Objective

This is the second time for JLU-NBBMS to join the iGEM competition, and compared to last year, this year we have more new iGEMers joining our team. At the beginning of the project, our team leader introduced iGEM competition and the application and prospect of synthetic biology to new iGEMers, and reviewed the ins and outs of last year's project together. On the one hand, members were encouraged to brainstorm for new inspirations, and on the other hand, to emphasize that the existing strengths of the lab serve as the cornerstone for new inspirations. In order to determine the theme of the new year's project, we discussed with the PI of the competition, Professor Zhang Ling from the Department of Pathophysiology, College of Basic Medical Sciences, Jilin University, about the current cutting-edge diagnostic and therapeutic methods for cancer, and attempted to find a feasible, innovative and practical solution for the project.

Contribution

During first meeting, Prof. Zhang Ling, the team PI, pointed out that although some cutting-edge approaches of cancer treatment were promising, they did not belong to the category of Synthetic Biology (e.g. Immunotherapy which used monoclonal antibodies combined with immune checkpoint inhibitors in tumor cells). And due to the limited time and knowledge reserve of the team members, she suggested that we consider attenuated Salmonella which was already available in the Pathobiology Research Laboratory of the College of Basic Medical Sciences in Jilin University as the chassis and based on last year's project (i.e. attenuated Salmonella in combination with RNAi technology), review the literature and analyze the projects of other iGEM teams. Combined with the current dilemmas in cancer treatment, we would like to analyze the pros and cons of last year's project for further exploring the potential of Salmonella to fight against tumors.

Implementation

Through talking with the PI, we learned that Bacterial-Based Cancer Therapy is a cutting-edge therapy in tumor treatment. Since Salmonella has good immunogenicity and targeting ability towards tumor, it can stimulate the autoimmune system to attack the tumor cells. Also its genetic materials are simple so it is easy for gene modification, and it can be used as a good vector for plasmids. However, it is difficult to be applied in clinical treatment due to its weak ability to kill tumor tissues by itself and its safety is hard to be guaranteed. So we decided to continue to use attenuated Salmonella mediated Bacterial-Based Cancer Therapy as a chassis to look for ways that we can further modify it.

Outlook

The next step is to think about how to further modify attenuated Salmonella to improve its safety while increasing its ability to kill tumor, analyze the current dilemmas in oncology treatment and look for new ideas by reviewing the literature and learning from the results of other iGEM teams over the years.

Objective

After identifying using attenuated Salmonella as the chassis of the project, in order to add innovation and social significance to make the most of our attenuated Salmonella to solve the challenges that arise in the treatment of cancer and solving the problems faced by cancer patients in a better way, we began to browse through the sharing of other teams on the iGEM blog.

Contribution

We learnt from the iGEM blog that microbial resistance is a very important area which points to a common public health challenge around the world. And we've seen many teams making amazing progress in addressing anti-microbial resistance, which is undoubtedly what every iGEMer wants - using Synthetic Biology to solve practical and urgent problems in real world, and to ‘make our world a better place’ through our efforts.

Implementation

After seeing the exciting presentations and results of many teams focusing on microbial resistance, we realized that the emergence of resistance is by no means limited to the microbiological field. In the field of oncology, the occurrence of drug resistance is also a major cause of poor efficacy of chemotherapeutic agents and short survival time. Therefore, we decided to address drug resistance as one of the goals of this year's project.

Outlook

We hope to take a step further on literatures of drug resistance, including the mechanisms of action in conventional chemotherapeutic agents as well as drug resistance in tumor cells, such as phenotypic changes, formation of inhibitory tumor microenvironments, and the changes of drug targets. Next we would like to further think about how to use Synthetic Biology to solve a specific problem and look for opportunities to bind it with attenuated Salmonella.

Objective

In order to learn about the latest advance on drug resistance in tumor, we carried out extensive literature reading, starting from internationally renowned journals such as Science and Nature. Then we divided into small groups for literature discussion and report for brainstorming, in the hope of absorbing advances and academic perspectives in the most cutting-edge researches. It laid a good theoretical foundation for our project.

Contribution

During the brainstorming session, we found that the research of drug resistance is a very important area. In 2005, Science posed 125 most challenging scientific questions in its 125th anniversary issue, and one of them was whether we could overcome drug resistance. Now almost 20 years later, more and more innovative therapies are on the horizon, but addressing drug resistance in cancer treatment remains the direction of efforts. Many new approaches are emerging continuously, such as Immune Checkpoint Inhibitor therapies that target immune evasion from tumor, and the use of gene editing techniques to repair drug-resistant genes in tumor cells. They all exert as a powerful force in the fight against drug resistance. Meanwhile, with the rapid development of computer technology, identification of drug-resistant genes in tumor cells through high-throughput sequencing and other techniques provides new targets and therapeutic strategies for the treatment of tumor with drug resistance.

Implementation

Since attenuated Salmonella has easy gene editing properties and targeting ability towards tumors, we believed it has the potentials of addressing the drug resistance as we learnt before. So we initially decided to use attenuated Salmonella to address resistance of chemotherapeutic agents used in tumor treatments due to its easy gene editing properties and its ability to target tumors.

Outlook

After initially defining the direction of our research, we decided to further learn the prevalence of drug resistance in different cancers and its impact on society in order to clarify our objectives. Next, we will go into hospitals to conduct interviews and communicate with cancer patients, doctors and other stakeholders to further consider the societal implications of addressing resistance in chemotherapeutic agents.

Objective

Through our engagement with the iGEM blog and science125, we initially recognized the importance of drug resistance research. But how much does drug resistance affect society? As Harlow Shapley noted, “Theories crumble, but good observations never fade”. To better understand how drug resistance affects treatment and the challenges patients face, we decided to conduct interviews with patients and their families at the Third Bethune Hospital of Jilin University (China-Japan Union Hospital).

Contribution

The struggles of patients and their families deeply moved us and strengthened our resolve to find solutions to chemotherapy resistance. Particularly, Patient 2's insights led us to reflect on the limitations of current treatments and sparked fresh thinking about the root causes of resistance. This feedback not only provided invaluable clinical insights but also inspired us to seek sustainable solutions. Our research promises to offer a new and effective pathway in cancer treatment, especially for hard-to-treat types of cancer. By employing innovative drug combination strategies, we aim to maximize therapeutic benefits while minimizing patient suffering.

Family Member 1’s feedback shaped our research direction, leading us to focus on the safety and efficacy of BBCT in reducing chemotherapy resistance. (Click here to see Design Safety.) We also became aware that many of the imported chemotherapy drugs used in treatment are not covered by health insurance, prompting us to explore health insurance policy reform.

Implementation

Safety has always been our priority, and privacy is no exception. Before conducting any interviews, we thoroughly consulted with participants and assured them that the interview content and findings would be used solely for research and statistical purposes, and would never be shared with third parties or used for commercial purposes(Click to view Safety ). Once consent was obtained, we signed a bilingual Interview Informed Consent with each participant. To safeguard the privacy of patients and their families, we will refer to them in this report using numerical identifiers.

We began with a detailed interview with Family Member 1, whose loved one is a gastric cancer patient. Family Member 1 revealed a concerning reality: with each chemotherapy cycle, resistance to the drugs gradually develops, leading to reduced efficacy, accelerated disease progression, and a reduced life expectancy. They were particularly concerned about how to slow down the onset of chemotherapy resistance to maintain the treatment’s effectiveness and prevent uncontrollable deterioration. They also shared that, to minimize suffering, imported chemotherapy drugs with fewer side effects and better results are often chosen, although these medications are expensive and not covered by health insurance.

At the same time, we introduced our plans for Bacterial-Based Cancer Therapy (BBCT) to Family Member 1. They expressed hesitation, noting that the word “bacteria” inherently evokes associations with disease. Whether BBCT would be considered as a treatment option would depend on how trustworthy and safe the process appears to patients and their families, as well as whether its safety can be verified.

Next, we interviewed Patient 2, a breast cancer patient. Her story highlighted the dual challenge of chemotherapy resistance: not only was it increasingly difficult to control the disease, but the resistance was also causing significant harm to her body’s normal tissues. Multiple relapses weighed heavily on her physically and emotionally. After switching to a new chemotherapy regimen, the cost of treatment skyrocketed, and the combination of drugs further weakened her body's tolerance, leaving her anxious and uncertain about the future. She expressed a strong desire to reduce drug resistance.

When we introduced BBCT to Patient 2, she was hopeful. Given her struggles with resistance, she expressed willingness to try the therapy if it proved effective. Patient 2 noted that late-stage treatments often deviate from established guidelines, explaining, “At this point, you just try this and that.” For emerging therapies, patients like her are primarily concerned with efficacy. Of course, because advanced-stage patients endure significant suffering, their tolerance levels and willingness to pursue treatment also influence their decisions.

Throughout the interviews, we expressed deep empathy and sincere care for both patients and their families, recognizing them as brave and resilient individuals on the frontline of the battle against cancer. As iGEM team members specializing in oncology, we deeply understand the immense challenges cancer treatment poses. The patients and their families warmly supported our efforts as medical students, acknowledging both our dedication to exploring cancer treatments and the humanistic care we provided.

Outlook

We hope this research will provide patients with more durable and stable treatment regimens, allowing them to manage their disease effectively for longer periods while improving their quality of life. By reducing chemotherapy resistance, future cancer treatments will become increasingly personalized and precise, not only enhancing treatment outcomes but also significantly alleviating the physical and psychological burden on patients. Additionally, we believe BBCT has the potential to become a breakthrough innovation in overcoming chemotherapy resistance.

Objective

To gain a deeper understanding of the mechanisms behind cancer drug resistance and its broad impact on clinical treatment, we had an in-depth discussion with Dr. Tianwei Wang, Deputy Director of the Comprehensive Interventional Oncology Ward at the Third Bethune Hospital of Jilin University (China-Japan Union Hospital). As a frontline clinician, Dr. Wang possesses extensive experience in cancer treatment, particularly in addressing resistance to chemotherapy and targeted therapies. His insights will help us understand the challenges from both the physician’s and patient’s perspectives. Through this exchange, we hope to further clarify the potential of BBCT in addressing drug resistance and provide guidance for translating our research into clinical practice.

Contribution

Dr. Wang’s data and insights confirmed the importance of our research focus, particularly regarding BBCT. His feedback led us to further refine our design to better meet clinical needs, such as improving bacterial targeting and enhancing therapy safety. These clinical insights provided both theoretical support and practical guidance, allowing us to be more focused in our research direction. We believe this study will ultimately provide more effective treatments for drug-resistant cancers, improving both patient compliance and survival outcomes.

Implementation

Dr. Wang shared critical data about cancer drug resistance, noting that over 90% of cancer-related deaths are associated with drug resistance. Certain cancers, like small cell lung cancer, exhibit resistance rates—both primary and acquired—exceeding 60%, while pancreatic cancer has a primary resistance rate of up to 70% for chemotherapeutic agents such as like gemcitabine. These figures underscore the magnitude of the drug resistance challenge in cancer treatment.

Dr. Wang further explained that resistance mechanisms and rates of resistance vary significantly across different cancer types. For instance, approximately 50-60% of non-small cell lung cancer patients treated with EGFR-TKI develop resistance due to the T790M mutation, while in melanoma patients, around 50% of resistance is closely related to NRAS mutations. These findings highlight the growing importance of developing personalized, targeted therapies to combat resistance in various cancer types.

Based on this discussion, we decided to focus on mitigating chemotherapy resistance by using BBCT to slow the progression of resistance.

Outlook

As research on BBCT progresses, we are confident that this innovative therapy will lead to significant breakthroughs, particularly in overcoming resistance to chemotherapy and targeted therapies. By delaying the onset of drug resistance, BBCT offers cancer patients a promising new treatment option that will not only control their disease more effectively but also enhance their quality of life.

Objective

The premise of any experiment and research is safety. After establishing the theme of using attenuated Salmonella to reduce chemotherapy resistance and understanding the high importance that clinicians and patients place on the safety of new therapies, we decided to first focus on modifying the safety of Salmonella first. We found that attenuated Salmonella is divided into many different strains due to its distinct characteristics. So, which one is more suitable as a chassis for resistance gene editing? To address this issue, we consulted Professor Yang Ming and Professor Dai Enyong.

Contribution

The Yang Ming team from the School of Basic Medicine at Jilin University provided us with a delayed lysis Salmonella strain χ11802. This bacterium's essential genes for synthesizing the peptidoglycan layer, asd and murA, are controlled by an arabinose promoter. In the absence of arabinose, the bacterial peptidoglycan layer cannot be synthesized, leading to automatic lysis. Since arabinose does not exist in the human body, we can control the lysis time of the strain by regulating the amount of arabinose carried, thereby ensuring the safety of the attenuated Salmonella. (link to safety)

During the discussion with Professor Dai, he mentioned that different tumor microenvironments significantly affect the targeting accuracy of bacterial vectors. After understanding the details of our project, he believes that using Salmonella as a vector is an excellent choice. Although there is heterogeneity among different tumor microenvironments, they share the common characteristics of low pH and low oxygen levels, which are completely consistent with the facultative anaerobic characteristics of Salmonella. Professor Dai also mentioned the currently popular oncolytic virus therapy, but it still has issues such as high immunogenicity, insufficient oncolytic efficacy, and systemic viral delivery obstacles. After understanding our bacterial vector which can simultaneously carry RNAi tools and chemotherapy drugs, Professor Dai believes that our project has good prospects, and he also suggested that the project could consider exploring the direction of immunotherapy.

Implementation

According to our conversation with our PI, we learned about the delayed lysis strain. We verified the feasibility of delayed lysis through literature search and experimental validation, deciding to use the delayed lysis Salmonella strain as our vector. The genetically engineered attenuated Salmonella can stably exist in the host and effectively deliver chemotherapy drugs and genetic engineering tools to specific tumor tissues without immediately triggering a lysis response. When the arabinose we provide is exhausted, the Salmonella will lyse and will no longer survive in the body, ensuring the safety of our system.

Outlook

Through discussions with Yang Ming and Professor Dai Enyong, we have gained a clearer understanding of the main components of the project. After listening to Professor Dai's suggestions regarding the tumor microenvironment, we began to further contemplate how to utilize the characteristics of the tumor microenvironment to minimize the impact of its variability on targeting accuracy and further improve the targeting accuracy of the Salmonella vector towards tumor cells.

Objective

After deciding to use Professor Yang Ming's strain, we learned that attenuated Salmonella still has certain toxicity to the human body, therefore if we want to use it as a vector, we must further reduce its toxicity through certain methods. Our team's PI Professor Zhang Ling from Jilin University's Pathophysiology Laboratory has done extensive research on the attenuation of Salmonella. To address the issue of Salmonella toxicity, we discussed with Professor Zhang.

Contribution

Professor Zhang suggested that we use genetic engineering techniques to precisely knock out the msbB gene closely related to Salmonella toxicity, which aims to weaken the potential harm of Salmonella to normal cells, with the hope of achieving a tumor treatment strategy that is both safe and effective. She emphasized that it is possible to retain some toxicity of Salmonella, because as a natural pathogen, its toxicity mechanism includes the ability to invade and destroy host cells; these characteristics, when properly regulated, can be transformed into a powerful tool against tumor cells.

Implementation

We were greatly inspired by Professor Zhang Ling's suggestions and, after discussion, reconsidered our understanding of the toxicity of Salmonella. We reviewed a large number of papers and found that retaining some toxicity of Salmonella does not cause serious secondary harm to the infected individuals. On the contrary, it can further stimulate the function of the immune system, mobilizing the body's own immune cells to attack tumor cells. At the same time, in conjunction with Professor Yang Ming's strain, we intend to directly modify the strain he provided by knocking out the msbB gene.

Outlook

After this communication, we have formally identified two aspects for modifying Salmonella for safety, one is to reduce toxicity, and the other is to utilize Professor Yang Ming's delayed lysis characteristics. After completing the control of toxicity and lysis time, we hope to achieve a further upgrade in safety by increasing the targeting ability of attenuated Salmonella.

Objective

In order to enhance the targeting ability of attenuated Salmonella, we reviewed a large amount of literature and identified many potential methods (targeting specific cytokines, expressing tumor-specific receptor binding proteins, etc.) To determine which method is the most feasible, we consulted Professor Quan Chengshi, who has conducted research in this area.

Contribution

We have gained deep insights from our esteemed teacher about how modern biotechnology has advanced to the stage where bacteria can be finely engineered. This innovative approach aims to significantly enhance the bacteria's ability to target and attack specific tumor tissues, opening up a novel and promising avenue for cancer treatment. He suggested that we use RGD peptide from the options we provided.

Implementation

After reviewing the information on RGD peptide and confirming its feasibility, we decided to follow the advice of our teacher and adopt RGD peptide to enhance the targeting capability of attenuated Salmonella. The RGD peptide can bind to integrin proteins that are highly expressed in tumor tissue, enhancing tumor-specific targeting ability of the bacteria. To further increase the ability of bacteria as 'vehicles' to target tumors, we utilized the Lpp-OmpA structure to display the RGD peptide on the outer membrane of Salmonella. OmpA is a bacterial outer membrane protein that tightly connects the RGD peptide to the outer membrane of Salmonella through the 'Lpp-OmpA 'structure.

Outlook

Our main vector, attenuated Salmonella χ11802, has been fully characterized. Next, we will focus on further enhancing the safety and specificity of the system. Specifically, we will verify and optimize the display effect of RGD peptide on the outer membrane of Salmonella through refined experimental design, aiming for more precise tumor targeting. At the same time, we will actively explore and validate different types of RGD peptides to select the most suitable peptide segments for our system, thereby further enhancing the therapeutic effect.

Objective

After considering the suggestions of the honest teacher, and also to achieve the goal of improving targeting capability of the system, as the wet experiments progress, we have decided to use the outer membrane expression system to present the RGD peptide. Through reading the literature we found that, with the in-depth research of other scholars, RGD peptide has gradually evolved from the initial common RGD peptide to various functionally modified RGD peptides. In order to screen for the most suitable RGD peptide for our system, we conducted experiments comparing the affinity and stability of different RGD peptides in binding to integrins in order to apply it.

Implementation

We obtained the protein structure of integrins from the Uniprot database, which contains two subunits: the alpha chain and the beta chain. At the same time, we used Alphafold to construct the structural files of three fusion proteins: Lpp - OmpA - RGD, Lpp - OmpA - RGDFK, and Lpp - OmpA - iRGD. After obtaining the protein structures of the two types of proteins, we simulated the molecular docking process using PDBePISA and Alphafold3. We concluded that Lpp-OmpA-RGD has the highest affinity.

Outlook

After testing, we believe that unstable fusion proteins may lead to lower reliability in our subsequent molecular docking and molecular dynamics simulations. To address this issue, we hope to improve the structure of the fusion proteins in the future, or use previously validated structures to enhance the stability of the fusion proteins, thereby improving the safety and specificity of our system. 【Link to experimental cycle 2】

Objective

After targeting based on the integrin protein on the surface of tumor tissue, we attempted to consider utilizing the internal microenvironment factors of the tumor to further enhance the safety and specificity of the system. Therefore, we invited Professor Sun Wei from the School of Basic Medicine at Jilin University for a discussion, hoping to gain insights on how to improve the safety and specificity of the system during our exchange.

Contribution

Professor Sun Wei conducted an in-depth analysis of the significant differences between the tumor microenvironment and the normal tissue environment, revealing the complex ecology of tumor growth and development. He pointed out that the tumour microenvironment differs significantly from the normal tissue microenvironment in several key aspects. Firstly, oxygen concentration is one of the notable distinctions; due to the abnormal angiogenesis and disorganized structure within tumor tissue, blood supply is insufficient, leading to generally lower oxygen levels in tumor regions compared to normal tissue. This phenomenon is referred to as the hypoxic tumor microenvironment. This hypoxic state not only affects the metabolic pathways of tumor cells but also promotes their invasiveness and enhances drug resistance. Professor Sun Wei also emphasized that the pH value of the tumor microenvironment is significantly lower than that of normal tissue. This is because the rapid proliferation of tumor cells is accompanied by a sharp increase in metabolic activity, producing a large amount of acidic metabolites, such as lactic acid, which accumulate in the tumor tissue, causing a decrease in local pH and forming an acidic microenvironment. This acidic environment not only favors the survival and proliferation of tumor cells but may also damage the surrounding normal tissue, further exacerbating the malignant progression of the tumor.

Implementation

Attenuated Salmonella itself has a natural advantage in targeting hypoxic and acidic environments. We hope to further utilize the characteristics of the bacteria and the value of the tumor microenvironment, considering whether we can leverage the special internal conditions in these tumor tissue microenvironments to trigger the expression of relevant effector genes. Through our research, we found that by using the hypoxia-inducible promoter pnirB, which responds to the hypoxic conditions in the tumor microenvironment, we can ensure that Salmonella expresses relevant effector genes under hypoxic conditions, further enhancing the specificity and safety of the treatment.

Outlook

Professor Sun Wei's profound insights into the uniqueness of the tumor microenvironment provide us with new research directions. In particular, we plan to further validate the role of the hypoxia-inducible promoter pnirB in our experiments. Through this validation process, we aim to confirm whether pnirB can effectively activate the expression of Salmonella-related effector genes under the hypoxic conditions of the tumor microenvironment, thereby not only enhancing the specificity of the treatment but also further ensuring its safety.

Objective

Having identified tumour drug resistance as the main direction of research in this project and performed the basic strain modification, we obtained the ideal vector, safe and targeted Salmonella. Next, we will explore how to use it to reduce tumor drug resistance. In order to find the method for knocking down drug resistance genes in this project, we conducted many brainstorming sessions. After literature review and group discussion of all members, we initially identified three knockdown gene strategies, namely: RNAi, CRISPRi and circRNA sponge. Next, we discussed with our PI Professor Yang Ming and Professor Zhang Ling. Professor Yang Ming is an expert in molecular biology and can provide us with his insights on gene knockdown methods and assess feasibility. We hope to finally determine the gene knockdown method through the discussion with the professors, which will lay the foundation for our subsequent experiments.

Contribution

In the process of brainstorming, we understand through the literature circRNA sponge principle is to use circular RNA as a competitive inhibitor of miRNA, thus indirectly improving the sensitivity of anti-tumor drugs, but we found through the literature artificial transfection circRNA expression plasmid, the cell first formation of linear RNA ring rate is not higher than 20%, knockdown miRNA efficiency is not high, so we no longer consider circRNA sponge.

Our team members continued to brainstorm times with PI about how to choose between RNAi and CRISPRi. Professor Yang Ming proposed that although CRISPRi has low off-target rate and high silencing efficiency, the sequence length of CRISPRi expressing CRISPRi is longer, and our other modification elements may cause the plasmid length to exceed 1000 kb. If the sequence expressing CRISPRi is placed individually into a vector to transfer two plasmids into the bacteria, plasmid incompatibility is likely.

Implementation

After the discussion, we further reviewed the literature and found that RNAi mediated gene silencing through small interfering RNA (siRNA), which can effectively reduce the expression of specific drug resistance genes. Considering that the laboratory where the team is located has mature experience in RNAi technology, if CRISPRi is selected, it will require a long time to explore the technology, which is not conducive to the construction and efficacy verification of the system in a limited time. Therefore, we decided to prioritize RNAi methods to knock down individual resistance genes to ensure the smooth progress of the project, while laying the foundation for further optimization of the system at a later stage.

Outlook

After determining the method to knock down resistance genes, our next step was to design RNAi modules for screening resistance genes, which also faced arduous challenges, so we began to plan the subsequent model design and consider how to improve.

Objective

After determining the knockdown of resistance genes using RNAi, we first began to design the RNAi module for Salmonella, but encountered two problems in the design process: 1. How to narrow the range of differential genes; 2. How to select appropriate indicators to rank the correlation between resistance genes and prognosis.(Link to model cycle1 )

Therefore, in the design process, we consulted our teacher, Mrs. Qi Mingran, hoping that after communicating with her, we could find an appropriate method to narrow the scope of screening for drug resistance genes and determine the appropriate indicators to rank the correlation between drug resistance genes and prognosis.

Contribution

At the beginning, by reviewing the literature, we found that we could use public databases to extract the drug use of patients, classify patients, and screen differential genes for further prognostic analysis.

Therefore, we plan to obtain a large number of tumor clinical data and gene expression data from TCGA and GEO databases. After classification according to drug use, we will screen the differential genes and conduct enrichment analysis on the differential genes to initially determine their functional changes, and conduct prognostic analysis.

However, after testing, we found that there were too many selected differential genes, which were not suitable to guide our personalized treatment. If we could narrow down the range of selected differential genes, it would be more conducive to our target screening. At the same time, it is particularly important to select the appropriate indicators to evaluate the relation of drug resistance genes and prognosis.

According to this, Mrs. Qi gave the corresponding suggestions, and we implemented and carried out the test.

Implementation

To reduce the selected scope of differential genes, we redesigned new parameters, tightened the expression level (here the normalized LogFC), and tightened the difference value of genes, and ranked the resistance genes according to the risk coefficient of the prognostic analysis as an indicator, as alternative targets for treatment.

After simulation, we obtained a more perfect stem experiment section of "Tumor resistance targeted gene screening and prognosis correlation analysis" 2, and obtained the therapeutic targets from the perspective of pan-cancer.(Link to model cycle 4 )

Outlook

After initially completing the design of the RNAi module, we planned to evaluate the utility and feasibility of the project next and consider how to better achieve resistance genes inhibition with RNAi. To this end, we contacted the BNUZH-CHINA 2023 team, who also applied RNAi last year, and discussed with them, hoping to get inspiration or some other suggestions (link to model cycle 3 ).

Objective

After intense brainstorming during the team meeting and discussions with our PI, Professor Yang Ming, we finally decided to prioritize the use of RNAi to knock down a single drug-resistant gene. While considering solutions to address tumor chemotherapy resistance, we are also seeking strategies to enhance the targeting specificity of Salmonella. Through extensive literature review, we have identified a targeting method distinct from environmental targeting, which is based on the structure of integrins on the tumor surface, namely RGD peptide-mediated targeting. In order to seek a stable RNAi expression method and assess the feasibility of RGD peptide targeting, we communicated with our old friends, BNUZH-China, to further improve and refine our project in terms of both drug resistance and targeting.

Contribution

In May, we had an online meeting with BNUZH-CHINA for an exchange of ideas and discussion. Focusing on drug resistance and targeting specificity, our team members actively participated in the discussion. We sequentially delved into in-depth discussions on both wet lab and dry lab.

The topics concerning wet lab mainly focused on what methods we should use to enhance the expression of RNAi to ultimately achieve our desired effect of knocking down a single drug-resistant gene. During our communication we learned that they used a bacterial-mediated RNAi, transkingdom RNAi (tkRNAi), in their project research. We discussed that the efficiency of gene suppression mediated by tkRNAi is higher compared to gene suppression mediated by using bacteria only as a vector for delivering shRNA expressing plasmids.

In terms of dry lab work, we delved into topics such as improving fusion proteins and the structure of Lpp-Ompa-RGD. We utilized the properties of RGD peptides to enhance the targeting capability of engineered strains through theoretical analysis and computational modeling. We are well aware that the structure of fusion proteins plays a crucial role in their function and application, making the improvement of this structure a core topic of our discussion. During the discussion, the BNUZH-CHINA team shared the protein structures they used in last year's project to us, which undoubtedly increased the possibilities for our team to utilize RGD peptide-mediated targeting. This provided us with valuable reference and inspiration.

Implementation

During the discussion, team members actively engaged in the conversation and believed that tkRNAi could better integrate into the gene circuits we designed. Additionally, by utilizing the T7 expression cassette within the tkRNAi module, bacteria could efficiently express other gene products, thereby expanding the application scope of the system. Consequently, we ultimately decided to use tkRNAi as the method to inhibit drug resistance genes.

At the same time, we also conducted structural simulation, molecular docking, and molecular dynamics simulations based on the main structure of the extracellular expression system. Using the protein structure of integrin retrieved from the Uniprot database, we utilized Alphafold to generate the structural files for the three improved fusion proteins: Lpp'-OmpA-RGD, Lpp'-OmpA-RGDFK, and Lpp'-OmpA-iRGD. We observed a significant improvement in the confidence level of the structures, although it did not reach an extremely high level. However, these structures have been experimentally validated and used by collaborative teams such as BNUZH-China. Therefore, we believe that these structures can support the credibility of molecular docking and molecular dynamics simulations. Subsequently, we conducted molecular dynamics simulations and molecular docking between integrin and the three improved fusion proteins. The results indicated that the Lpp'-OmpA-RGD, which we ultimately selected, exhibited the best binding effect.

Outlook

In terms of wet lab experiments, based on our discussions and implementations with the BNUZH-CHINA team, our next step is to consider how to utilize the T7 expression cassette to modify our engineered bacteria. Additionally, we will use flow cytometry to detect whether the RGD peptides are expressed on the surface of our engineered bacteria.

In the context of dry lab work, the team members have determined that the fusion protein Lpp'-OmpA-RGD exhibits the strongest stability. Moving forward, we will focus on optimizing the overall system design, with the aim of advancing the idea of engineered bacteria carrying nanoparticles encapsulated with chemotherapy drugs. To ensure that the practical application of this technology is both safe and effective, the team hopes to validate the safety of the system through modeling and design, as well as address the issue of nanoparticle synthesis. Since we are using bacterial therapy and have created new engineered bacteria, the team has made the decision to re-evaluate their safety. We plan to simulate the changes in the body's immune system after the injection of Salmonella, as well as the delayed lysis system to ensure its safety.

Objective

After thorough discussions with the BNUZH-CHINA team, we plan to modify our engineered bacteria to possess a complete T7 expression system. Initially, we attempted to express both the T7 promoter and T7 RNA polymerase through plasmids. However, we found that the length of T7 RNA polymerase is approximately 2.7kb. If inserted into the plasmid vector, it would cause the final plasmid size to exceed 10kb, leading to issues such as low efficiency during plasmid transformation. The team had several internal discussions but had not yet found an effective solution. Therefore, we once again consulted with our PI, Professor Yang Ming, to discuss how to resolve this bottleneck problem.

Contribution

Professor Yang Ming was appreciative of our plan to use T7 RNA polymerase for the modification of our engineered bacterial strain. He suggested that we insert the T7 RNA polymerase gene into the genome of Salmonella, while the T7 promoter is located on our designed pSilencer-target gene plasmid without integrating into the bacterial genome. This approach will help reduce the size of the plasmid and improve the efficiency of plasmid transformation.

Implementation

We inserted the T7 RNA polymerase gene at the location where the msbB gene was knocked out.

Figure 1. Our modifications to the χ11802 genome.

Outlook

At this point, the modifications to the engineered bacteria used in our project are basically complete. Overall, we used the delayed lysis strain χ11802 as our engineered bacterial strain. We knocked out the msbB gene to attenuate the toxicity of Salmonella, and leveraged the Lpp-OmpA structure to display the RGD peptide on the outer membrane of Salmonella, enhancing the tumor-specific targeting ability of the bacteria. We also employed the hypoxia-inducible promoter pnirB to respond to the hypoxic conditions in the tumor microenvironment, ensuring that Salmonella expresses relevant effector genes under hypoxic conditions. Finally, we inserted the T7 RNA polymerase gene in situ at the msbB knockout site and transformed the plasmid carrying asd, murA, and the T7 promoter (see Design for details). So far, we have partially addressed the issue of chemotherapy resistance. Next, we will refine our approach to maximize the impact of our project.

Objective

Our engineered bacteria have made significant progress in terms of resistance and safety. The project was reinitiated during a brainstorming session where Professor Yang Ming from the Jilin University School of Basic Medical Sciences proposed a core definition of engineered bacteria. Professor Yang believes that if the bacteria only contain RNAi segments, their modular characteristics are not significant and their innovation is insufficient to be truly termed "engineered bacteria." Given our team's initial goal of addressing chemotherapy resistance, this discussion led us to seek professional guidance from Dean Xin Ying. Interviews with patients and doctors deepened our understanding of their worry for chemotherapy side effects and the physicians' helplessness, prompting us to find ways to mitigate these effects.

Contribution

Dean Xin Ying, while approving of our project, raised a concern: "If chemotherapy drugs and modified Salmonella are injected together without any interaction, it doesn’t address the systemic side effects caused by chemotherapy nor fully utilize the bacterial vector. Is your project not maximizing its potential?" Dean Xin emphasized that the primary challenges in tumor chemotherapy are resistance and side effects. Addressing both could significantly alleviate patient suffering while enhancing therapeutic efficacy. This prompted us to rethink our project: could we enhance its tumor targeting to reduce the systemic toxic reactions of chemotherapy drugs?

Implementation

Inspired by Dean Xin Ying, our team explored two avenues: the conversion of prodrugs into active drugs through enzymes expressed by Salmonella, and the use of advanced materials to attach chemotherapeutics directly to the bacterial surface. As current research on prodrug conversion is more mature only for cisplatin, we chose for the latter—using novel materials to load chemotherapy drugs onto bacteria, thus broadening the experimental scope and enhancing specificity and efficiency of treatment.

Outlook

We aim to select the most suitable materials for carrying chemotherapy drugs so that Salmonella can deliver these drugs directly to tumor tissue more precisely, significantly reducing impact on normal cells while increasing the precision and safety of treatment. This not only optimizes existing chemotherapy methods but also opens new possibilities for future cancer treatments. We hope to create a truly interchangeable, iterative drug delivery system in the form of engineered bacteria, that will benefit a broader population.

Objective

We are focused on developing an innovative drug delivery system using engineered Salmonella as a carrier to precisely deliver chemotherapy drugs to tumor tissues. The core of this method lies in selecting the most suitable loading materials to ensure effective drug delivery to the tumor site while minimizing damage to normal cells. In selecting materials for carrying chemotherapy drugs, we consulted with Professor Xu Cai Na from the Department of Biochemistry at Jilin University School of Basic Medical Sciences to discuss the feasibility of our ideas.

Contribution

Professor Xu Cai Na suggested using nanotechnology to encapsulate chemotherapy drugs, enhancing drug targeting through precise delivery. This greatly advanced our understanding of the drug delivery system and highlighted its potential in reducing drug side effects and improving therapeutic outcomes.

Implementation

Our literature review revealed various methods for attaching drugs to bacteria, including chemical bonds, ionic bonds, or biotin. In-depth discussions with Professor Xu Cai Na revealed that chemical bonding offers stability and feasibility. We decided to try linking attenuated Salmonella and nanoparticles through chemical bonds and chose PEG-PLGA nanoparticles. These particles can not only carry chemotherapy drugs but also connect via the abundant carboxyl groups on the bacterial surface, thus achieving targeted delivery.

Outlook

We aim to reduce chemotherapy resistance and side effects through this innovative drug delivery method, providing patients with safer and more effective treatment options. Learn more about how our innovative drug delivery system reduces chemotherapy side effects by clicking this link. [Insert Link Here]

Objective

After deciding to use nanoparticles to carry chemotherapy drugs and achieve targeted delivery on the surface of Salmonella, we urgently needed in-depth research and practical support in nanoparticle technology. To implement this innovative treatment strategy, we collaborated with the Chemistry Department at Jilin University to explore and develop nanoparticles suitable for our project.

Contribution

The Supramolecular Structure and Materials Lab at the Jilin University Chemistry Department, with its extensive background in nanotechnology, provided strong research support for our project. The lab's advanced equipment and expertise allowed us to conduct precise experiments and optimizations in nanoparticle design, synthesis, and functionalization.

Implementation

The design and synthesis of the PEG-PLGA nanoparticles was carried out by experts in the Chemistry Department with experience in nanotechnology. Due to their biocompatibility and biodegradability, these particles became the ideal drug delivery carriers. Our team worked closely with them, continually optimizing the particles' size and surface characteristics to suit various drug loads and targeting needs. To enchance the nanoparticles' targeting capabilities, researchers at the Chemistry Department developed a range of surface modification techniques, including the addition of specific targeting molecules like antibodies or small molecule ligands that can recognize and bind to receptors unique to tumor cells. Our team was involved in the design and implementation of these modifications, ensuring that these functional treatments not only enhanced targeted delivery but also preserved the drug's activity. Coupling nanoparticles with engineered Salmonella was one of the innovative aspects of this project. Our team led the development and testing of this coupling process, including assessing the stability and efficiency of nanoparticle binding with Salmonella. Through these experiments, we not only improved the bioavailability of drug-loaded nanoparticles but also optimized their distribution and tumor targeting in model organisms.

Outlook

Our multidisciplinary collaboration with the Jilin University Chemistry Department not only significantly enhanced interdisciplinary integration in our project but also provided a solid experimental foundation and prospects for potential clinical translation. We look forward to these research outcomes eventually transforming into practical medical solutions, bringing more precise and safer treatment options to chemotherapy.

Objective

After determining the drug delivery method of chemically binding nanoparticles, we decided to begin experimental verification of the effect of BIOTARGET on drug resistance in chemotherapy-resistant cancer cells. First, we need to find a cancer to determine the cell line for experiments. Therefore, we consulted Professor Enyong Dai.

Contribution

Professor Dai emphasized that ovarian cancer and breast cancer are two typical chemotherapy-resistant cancers, and the appearance of drug resistance during chemotherapy treatment greatly limits the quality of survival of patients. Both breast cancer and ovarian cancer are sensitive to chemotherapy, especially ovarian cancer, which is very sensitive to chemotherapy and does not belong to natural drug resistance. Therefore, it is an ideal drug resistance model. Based on these facts, we focused on specific cell lines of ovarian cancer and breast cancer. We consulted with Professor Dai and learned that in clinical practice, ovarian cancer is mainly treated with platinum-based drugs, so we selected human ovarian cancer cisplatin-resistant cell lines SKOV3/CDDP and A2780/CDDP. Breast cancer is mainly treated with 5-FU, ADM, and DDP, so we selected the human breast cancer multidrug-resistant cell line MCF-7/MDR for subsequent experimental verification, which can truly reflect the drug resistance challenges encountered in clinical treatment.

Implementation

After discussion with Prof Dai and review of literature, we decided to use the MCF-7/CLDN6 cell line and the drug-resistant cell line MCF-7/MDR to verify whether BIOTARGET can knock down target genes efficiently, and to verify whether BIOTARGET plays a role in the resistance of chemotherapy-resistant cells in the MCF-7/MDR, SKOV3/CDDP, and A2780/CDDP cell lines. After BIOTARGET specifically knocked down CLDN6, the sensitivity of breast cancer multidrug-resistant cell line MCF-7/MDR to three different chemotherapeutic drugs increased; When BIOTARGET was incubated with MCF-7/MDR, the expression of drug resistance-related proteins in tumor cells decreased; The combined use of ADM and BIOTARGET can greatly increase the early apoptosis rate of drug-resistant tumor cells; The sensitivity of SKOV3/CDDP and A2780/CDDP cells to DDP before and after the addition of BIOTARGET was determined by CCK8 assay, which verified that BIOTARGET could reduce the drug resistance of SKOV3/CDDP and A2780/CDDP cells; The co-culture of BIOTARGET and SKOV3/CDDP cell lines resulted in the highest rate of early apoptosis. After the addition of BIOTARGET, the expression of apoptotic proteins increased and the expression of anti-apoptotic proteins decreased.

Outlook

Due to time constraints, we only verified the effect of BIOTARGET on the drug resistance of chemotherapy-resistant cells in three cell lines: MCF-7/MDR, SKOV3/CDDP, and A2780/CDDP, covering only two types of cancer: breast cancer and ovarian cancer. We will further validate more types of drug-resistant cell lines in our future work, with a view to further validate therapeutic effect of BIOTARGET . We hope to use SKOV3/CDDP, A2780/CDDP, and MCF-7/MDR cell lines to verify the high efficiency of BIOTARGET knockdown of target genes and to verify that BIOTARGET can reduce tumor cell drug resistance.

Objective

After communicating with Dr. Enyong Dai, we realized that traditional tumor chemotherapy still has many obstacles: drug resistance, lack of selectivity, and can cause damage to normal cells, resulting in a series of side effects. Therefore, we hope to understand the patients' true thoughts about our new treatment to discover more areas where our project can be improved to be closer to clinical treatment. Therefore, we communicated with Patient 2.

Contribution

Before having a conversation with breast cancer patient 2, we signed an informed consent form with him(see safety). During the conversation, we asked him to rank the various aspects he was interested in of the new treatment in the form, and we captured the two core issues that patients are most concerned about regarding the new treatment --side effects and safety. Patients are generally concerned that the new treatment involves the introduction of bacteria, a non-traditional treatment method, which may pose unknown and unpredictable risks, such as uncontrollable infections or other undefined side effects. This feedback not only reflects the patients' expectations and concerns about the new treatment, but also provides the direction for our subsequent research and optimization.

Implementation

We reviewed previous experiments and integrated and sorted out measures to ensure safety and reduce side effects: we have knocked out the msbB gene through genetic engineering technology, which significantly reduces the toxicity of Salmonella and reduces its potential pathogenic risk. At the same time, we designed a strategy of delaying the lysis of the bacterial strain to ensure that the bacteria can self-destruct after completing their task, avoiding long-term presence in the host. In addition, we also introduced RGD peptides to enhance the targeting of bacteria to tumor tissues and reduce damage to non-target tissues. More importantly, we utilized the hypoxia-inducible promoter pnirB, which is an intelligent regulatory mechanism that allows Salmonella to specifically express therapeutic genes in the tumor's hypoxic microenvironment, further improving the precision and safety of treatment. Therefore, we believe that the feasibility of the BIOTARGET project is very high.

Outlook

When discussing the safety issues of Salmonella applications in depth, we sincerely invite all parties to join and share the latest breakthroughs and discoveries in Salmonella safety research, providing us with optimization ideas from different perspectives. This move aims to promote the project to be closer to clinical practice, more in line with patients' voices, and promote extensive exchanges and cooperation in the field. At the same time, it strengthens public science education, ensures the safe application and popularization of the technology, and lays a solid foundation for subsequent project optimization and improvement.

Objective

From July 12th to 14th, the 11th China Regional iGEMer Conference (CCiC) was held at Xi'an Jiaotong-Liverpool University. In order to further refine our project plan and present the best exhibition effect at the conference, we invited Professor Wang Fang, Dean of the College of Basic Medical Sciences at Jilin University, as well as our Project Investigators (PIs), Professor Zhang Ling and Professor Yang Ming, to conduct a comprehensive and systematic discussion on our project before departure.

Contribution

fter listening to our report, all three teachers provided constructive suggestions, which pointed out the direction for us to improve the project in the future. Dean Wang Fang pointed out the defects and loopholes in our dry and wet experiment designs. He emphasized the need to clarify "how to reflect personalized treatment" and noted that "compared to developing a new chemotherapy drug, our design of engineered iterative technology, will greatly benefit patients in both treatment efficacy and economic aspects, especially for those who cannot afford the high cost of targeted drugs, if it can reduce the issue of tumor chemotherapy resistance"(link to education ). He believed that our project had great potential, and our goal was not to solve tumor problems of all kinds, but as long as BIOTARGET could play a role in one type of tumor, everything we did would be meaningful. Professor Zhang Ling further clarified that the significance of modifying the original Salmonella chassis organism in improving efficiency. She also suggested that we collect epidemiological data on ovarian cancer and breast cancer. Professor Yang Ming believed that our dry experiment design was not complete enough, and he advised us to strengthen communication with other iGEM teams in this aspect at the CCiC.

Implementation

After the meeting, we combed the suggestions of the three professors. For the problem of personalized treatment, we designed shRNA that can be replaced according to the drug resistance gene status of different patients. We can also use other tumor-targeting peptides or antibody fragments instead of RGD peptides to enhance the tumor targeting ability of the treatment system and adapt to the treatment needs of different types of tumors. The suggestion of Dean Wang Fang brought us inspiration, and we have subsequently improved our investigation in the field of medical insurance. At the same time, we have further collected epidemiological data on ovarian cancer and breast cancer according to Professor Zhang Ling's suggestions, which is also a strong support for our entire project background.

Outlook

We have realized the deficiencies of the experimental part. In addition to bioinformatics analysis, we also need support in mathematical modeling. At the same time, we also need to think about how to integrate dry experiments and wet experiments into a whole, which is what we need to focus on at the CCiC.

Objective

Recognizing the significant challenges present in the implementation of our project, we sought more efficient solutions and opportunities for collaboration with other iGEM teams. Therefore, at the CCiC conference, we shared our project with iGEMers from the China and established connections with numerous teams.

Contribution

Before the CCiC, we had completed the basic design of our project and begun implementing our wet experiment plan. Although we encountered problems, we still found some solutions. Before departing, Professor Yang Ming's suggestions prompted us to consider how we could further improve our project design. With this confusion in mind, we engaged in deep exchanges with HUST-China, BNUZH-China, and HainanU-China at the CCiC. They shared their experiences in dry experiment design with us. We introduced the experimental details of nanoparticle preparation and linkage to BNUZH-China. During our discussions with them, we discovered that double-end modified PEG might affect the linkage between nanoparticles, leading to reduced material utilization.

Implementation

The communication with other iGEM teams has helped us to clarify our thoughts. Although CCiC has ended, we have kept in touch. Finally, our experimental part will be implemented in three parts: "Model screening and verification of BIOTARGET targeting function", "Screening of targeted genes for tumor drug resistance and prognostic correlation analysis", and "Analysis of safety and efficacy of BIOTARGET"(link to model ). In CCiC, we found the shortcomings of nanoparticles in the project. To address this issue, we quickly reviewed relevant literature and found that we need to redesign the nanoparticles and optimize their functionality. So how can we solve this problem? Our PI believes that we need to discuss this issue with professors from the College of Chemistry at Jilin University.

Outlook

After returning back, we sorted out through our preliminary thoughts and confusion about nanoparticle modification and are ready to discuss this issue with teachers from the College of Chemistry at Jilin University in the future.

Objective

The 11th China Regional iGEMer Exchange Meeting CCiC welcomed 89 university teams and 35 high school teams from China, which was a grand gathering. JLU-NBBMS hopes to take this opportunity to gather iGEMers from all over the country, unite our strength, and jointly explore more possibilities of human practice. Before the start of CCIC, JLU-NBBMS took the initiative to contact nearly 20 teams from the University of Macao, Beijing Normal University Zhuhai Campus, Wuhan University, etc., and sent them an invitation letter for HP&Eductaion intercollegiate cooperation. We hope to have in-depth exchanges with them on the project planning and development direction of HP and Education, and form a certain degree of cooperation relationship. We know that the strength of one party is weak, so we have the responsibility to call on more iGEMers to join the cooperation team of JLU-NBBMS, and gather the strength of everyone to attract more people.

Contribution

After the roundtable discussion began, the team members of JLU-NBBMS distributed an activity plan prepared in advance for the participating teams, which included our preliminary ideas for the "Biomedical Research Course", "Biosafety & Ethics White Paper", and "Bio-field Synthetic Biology Comic Book". For detailed information, please refer to the attached PDF. The detailed PDF will be attached at the end of the article. This is just a starting point, and we hope to gain more inspiration during the discussion and engage in thought exchange and collision. We first briefly introduced these three activities, expressing our original intentions and ideas, and then ushered in a heated discussion, during which we recorded everyone's feedback. After the discussion, many teams expressed their desire to form close partnerships with us, which gave us confidence to promote our activities in the future.

During this collaborative roundtable discussion, we received suggestions from various sources. Initially, we thought that the iGEM Biomedical Research Course iBSRC would only be open to students from Jilin University, but after listening to our presentation, other participating teams expressed their desire to expand the scope of enrollment to include not only Chinese university students, but also some high school students and even students from Malaysia. Therefore, publicity became one of the most critical aspects of our event. After the meeting, we reached a consensus with our partner team after careful consideration. During the discussion of the "Biosafety & Ethics White Paper", XJTLU-Software provided us with new inspiration, which prompted us to include AI-related ethical and security issues in the white paper. Finally, our plan for Bio-field synthetic biology comic strips was also recognized by everyone, but they suggested that the comic strip format is not conducive to dissemination on most social media platforms, where most users browse by page. This can disrupt the continuity of the comic strip reading experience, so we have conducted more in-depth discussions on the painting format in the future.

Implementation

After CCiC, we have integrated everyone's suggestions and improved the three projects to varying degrees, and gradually pushed forward the realization of the projects. Finally, JLU-NBBMS, ZQT-Nanjing, WHU-China, CAU-China, BNUZH-China, and PekingHSC jointly held the iBSRC-Middle School event, and with PekingHSC, BNUZH-China, WHU-China, we held the iBSRC-University event. And with XJTLU-Software, JLU-CP, and other 18 teams, we jointly wrote the "Biosafety & Ethics White Paper", and with JLU-CP, DUK iGEM, and other 8 teams, we completed the Bio-field synthetic biology comic series. (link to education )

Outlook

We are very much looking forward to the scene after the event. During this collaboration exchange night, we have become very good partners with many iGEM teams, which has laid the foundation for our rich exchange activities.

see our White Paper in Education

Objective

In our efforts to design nanoparticles, we encountered unique challenges. We discovered that dual-end modified PEG leads to self-linkage among nanoparticles, significantly reducing material utilization efficiency. To address this technical issue, we collaborated with Jinsei Pharma, renowned for its extensive expertise in chemotherapy pharmaceuticals, and engaged with Professor Zhang Yuetao, a leading figure in nanoscience from the Jilin University Chemistry Department. Our discussions focused on seeking breakthrough innovations and solutions for these challenges.

Contribution

Throughout our deep discussion with Jinsei Pharma, they introduced a visionary perspective: viewing Salmonella as a "natural linker," thereby avoiding the need for chemical linkers. This approach not only optimized material utilization but also simplified the design of the nanoparticles. Furthermore, under the guidance of Professor Zhang Yuetao, we fundamentally redesigned the functional architecture of the nanoparticles by directly attaching amino groups, effectively solving problems related to linkage efficiency.

Implementation

Inspired by these innovative ideas, we opted to employ multiblock supramolecular polymer technology for fabricating the nanoparticles. Specifically, we utilized the amino side chains of lysine as surface modifiers to ensure the presence of active amino groups on the nanoparticle surface. This strategy, based on the principle of charge repulsion, prevents self-linkage and enhances the efficiency of targeted delivery. To avoid premature phagocytosis in the bloodstream, we constructed the nanoparticles using a triblock copolymer of mPEG-PLGA-PLL, ensuring improved biocompatibility and circulatory stability within the body. In the Supramolecular Structures and Materials Laboratory at Jilin University's Chemistry Department, our team members, under expert supervision, successfully synthesized these novel nanoparticles and extensively characterized their morphology and size using transmission electron microscopy.

Outlook

Through this series of innovations and collaborative efforts, we have developed a highly targeted, safe, and efficient nanoparticle-based chemotherapy drug delivery system. Looking forward, we plan to utilize mathematical models to further investigate the synthesis dynamics and drug release behaviors of nanoparticles, simulating their in vivo effects. This study aims to evaluate whether this approach can achieve precise, targeted treatment goals and to contribute to technological breakthroughs in cancer treatment.

Objective

In the pursuit of precision medicine, the accurate synthesis of nanoparticles and control over their target drug release behavior are especially critical. To achieve this objective, we collaborated with the Mathematics Department at Jilin University to initiate a series of mathematical modeling salons, [Link to Educational Salon], aimed at enhancing the safety and efficiency of nanoparticle applications through interdisciplinary cooperation, and to lay a solid theoretical foundation for their use in medical fields.

Contribution

During the salon titled “Innovative Applications and Exploration of Mathematical Modeling in Medical Research,” Professor Song Haiming from the Mathematics Department presented foundational knowledge on mathematical modeling related to medicine to colleagues from the Basic Medical College. This expanded the perspectives of the Basic Medical College students and stimulated further thought on the establishment of our project model. In the latter part of the salon, we collaborated with Professor Song to design a mathematical model for system safety. Through simulation experiments, we detailed the synthesis process and drug release mechanisms of nanoparticles, ensuring the feasibility and safety of our designs. Moreover, these discussions and model developments provided quantitative guidance for our experimental design, significantly enhancing the predictability and success rate of our experiments.

Implementation

Based on the discussions and recommendations from the salon, we formulated a series of specific experimental protocols, which includes tests for the stability of drug carriers and evaluations of release efficiency. Through mathematical modeling, we were able to predict the behavior of nanoparticles under various conditions, optimize experimental conditions, and reduce the randomness and resource waste of the experiments.

Outlook

Through close collaboration with the Mathematics Department, our project has made further progress on the basis of theoretical and simulation experiments. In our future work, we plan to further deepen this interdisciplinary collaboration. By continuously comparing modeling results with experimental data, we will refine and optimize existing models based on feedback. In addition, we hope to extend the predictive capabilities of mathematical models to more areas of biomedicine, especially by introducing more precise gating mechanisms into nanoparticle drug delivery systems. This would allow drugs to be released only in specific tumor microenvironments, greatly reducing the impact on normal tissues, thereby alleviating patient treatment side effects and enhancing the therapeutic efficiency of drugs. Futhermore, we plan to explore the use of mathematical models to evaluate novel biocompatible materials to support broader applications of our nanoparticle systems.

Objective

In our mathematical modeling salon, insights gained prompted us to revisit and optimize our experimental designs, especially concerning target gene screening and drug delivery strategies. By collaborating with experts in mathematics and chemistry, we designed more accurate models to predict and verify the behavior and efficiency of drug carriers. This process not only increased the predictability of our experiments and reduced resource waste but also enabled more precise control of drug release behaviors in vivo. In the iGEM project, addressing the challenges from "Cycle 1," we implemented stricter criteria for target gene selection based on prognostic analysis-derived sorting standards. Combining challenges encountered in "Cycle 3" and improvements to the overall system design, we introduced an engineered bacteria scheme carrying nanoparticles wrapped in chemotherapy drugs. To advance this scheme, we collaborated with the Mathematics and Chemistry Departments at Jilin University to explore system safety modeling design, nanoparticle synthesis, and key drug release technologies.

Contribution

Our design focused on narrowing the range for selecting differential genes and redefined new parameters for gene expression levels (normalized LogFC) and differential values. Furthermore, based on risk coefficients from prognostic analysis, we re-ranked drug-resistant genes as alternative therapeutic targets. Regarding safety, we designed two crucial modules: (1) changes in the immune system following Salmonella injection and (2) simulation of a delayed lysis system. For nanoparticles, we concentrated on simulating factors influencing drug release.

Implementation

Through a series of simulation experiments, we optimized the experimental scheme for "Tumor Resistance Gene Targeting and Prognostic Relevance Analysis," obtaining pan-cancer treatment targets. The experiments primarily simulated the safe range of injections and the safety of delayed lysis. Ultimately, we integrated various part models to thoroughly analyze the overall system's effect, ensuring the practicality and safety of the scheme.

Outlook

Upon completing this phase of research, our three major sections, "BIOTARGET Model Screening and Validation," "Tumor Resistance Gene Targeting and Prognostic Relevance Analysis," and "BIOTARGET Safety and Efficacy Analysis," have successfully concluded. In the future, we will continue to verify the consistency of these models with actual experimental data and adjust and optimize the models based on experimental outcomes. Through this approach, we aim to further enhance the precision and efficacy of our therapeutic schemes and ultimately achieve personalized precision medicine solutions.

To expand our team's research scope and provide both technical and theoretical support, JLU-NBBMS members have broadened their international perspectives and honed their cross-cultural communication skills. We visited the University of Tokyo and Waseda University in Japan, where we engaged with local students and exchanged insights on our project with student associations, research groups, and professors at the University of Tokyo and Waseda University. Through campus interviews, surveys, in-depth discussions, and group dialogues, we gathered substantial data and feedbacks on our initiatives. Moving forward, JLU-NBBMS 2024 is committed to enhancing international cooperation, learning from each other's strengths and weaknesses, and integrating societal insights to enrich our research endeavors.

Objective

We aspire to travel abroad to Asia, Europe (see Animal Ethics for details) and many other places for exchanges of projects, listen to a diversity of opinions and optimise the design and ideas of our project.. Our goal is to delve into the insights of researchers in different regions regarding Synthetic Biology and Bacterial Based Cancer Therapy. This trip not only finished our team with invaluable research data, but also sparked brainstorming with researchers from different regions, generating novel ideas and technical support for our endeavors. In addition, during the trip we strengthened the communication with internationally renowned universities and research teams. We shared the experience of the JLU-NBBMS iGEM project, explored potential collaboration opportunities, and fostered international academic exchanges and cooperation.

Contribution

To our team, this activity was a rich source of insights. We not only collected a large amount of data on the views of students in Japanese university on cutting-edge biotechnology, but also required cutting-edge knowledge about the application of AI in biomedical field through lectures and seminars by this activity. Additionally, team members engaged in a productive diaogue with Prof. Wu, which helped to further clarify and improve the design of the study on‘The screening for targeted gene of drug resistance in tumour and prognosis correlation analysis’. This exchange provide a new direction and technical support for our research endeavors.

Implementation

Prior to the trip, our team established contact with the relevant departments and student organizations of the University of Tokyo and Waseda University through email, social media and other communication channels. This was done to specifically confirm the details of the visit and the arrangements for the activities. We completed preparatory work, including the design of the research programme, the creation of the questionnaires, interview appointments and cultural adaptation training to ensure the successful implementation of the activities.

On the campuses of The University of Tokyo and Waseda University, our team members interviewed more than 50 local students within two days. This allowed us to effectively gather insights into the attitudes and acceptance of Japanese university students towards the cutting-edge biotechnology such as oncology treatment and Bacterial Based Cancer Therapy.

Figure.1 Interviews with Japanese students at the University of Tokyo

Figure.2 Interviews with Japanese students at Waseda University

Table.1 Acceptance of Bacterial Based Cancer Therapy of the students in Japanese university students

At the same time, we engaged in in-depth conversations with the student associations and research teams of the two universities. We shared the design and experience of our project while exploring opportunities for cooperation.

Figure.3 Exchange with student clubs at the University of Tokyo

Team members attended lectures by professors from two universities. A professor from the University of Tokyo taught us about the application of medical AI in biomedicine.

Figure.4 Lecture at the University of Tokyo

Figure.5 Conversations with professors from the University of Tokyo

It is worth mentioning that the team members were greatly inspired by the lecture given by Professor Xianchao Wu of Waseda University. After the lecture, we discussed the design of our project with and explained our initial ideas for the dry experiment (see Model). Prof. Wu pointed out that grouping by medication status as a label can be used to pave the way for 'supervised learning'. He suggested that we could use this method to optimize the screening of drug-resistant genes through machine learning. If we had difficulties in writing the algorithm, he advised we could select from existing gene screening algorithms and choose the one with higher accuracy for our project. At the same time, our team members mentioned the problem that there are too many differential genes being screened in the process and Professor Wu suggested us to tighten the differential conditions in the screening process. He then pionted out that we could prioritize genes based on the prognostic risk coefficient index, which clarified the original ambiguity about the design. During the communication between the team members and Prof. Wu optimized and clarified parts of our design for the dry experiment.

Figure.6 Lecture by Professor Wu Xianchao of Waseda University

Outlook

We will continue to deepen our domestic cooperation and expand our exchanges and cooperation with foreign research institutions. The team will make full use of the valuable experience and resources gained from these events to further optimize our Model and strengthen the applied research of big data and artificial intelligence in biomedical field. In addition, the team will also focus on cultivating members' international vision and cross-cultural communication skills, thereby laying a more solid foundation for future international cooperation and exchange.

Objective

Once our team had completed the dry and wet labs, we turned to GeneRulor to analyze and evaluate the entire project in order to ensure a high and stable level of safety and feasibility in the realization of our experiments.

Contribution

Zhuhai Shutong Medical Technology Co., Ltd. is a biomedical enterprise with gene editing as its core technology, which is committed to carrying out new drug research and development for a variety of diseases, especially tumours, and providing gene-targeted therapeutic solutions through innovative technologies. The company has built a one-stop gene editing platform, a molecular diagnostic platform and a scientific and technological service platform, which can provide a full-process solution from target screening, gene editing effectiveness testing to safety assessment.and the fact that there is a precedent of successful editing of Salmonella before it makes our team excited toEvaluation of communication and co-operation with the company.

Implementation

After understanding our team's overall project vision and the need to assess safety and feasibility, the company applied one of its self-developed high-throughput off-target assays, AID-seq, to test the effectiveness of our knockout and found that our off-target rate was at a detection threshold of 0.2%, and that potentially off-target loci are considered safe.AID-seq can detect more low-frequency and true off-targets, which is critical for gene therapy. This is because even a small number of cells with off-target editing may lead to cancer due to clonal amplification. This is also a strong proof of the absolute safety of our project in terms of gene editing. In addition, based on the company's many years of experience in innovative research, the professionals also gave an optimistic assessment of the feasibility of the project.

Outlook

With the professional testing methodology and data support , we have more confidence in our project. The main body of the project is now almost finished, and we would like to take the complete project back to the closest stakeholders - doctors and patients - for further evaluation and refinement in a real-world clinical perspective.

To demonstrate whether our team's outcomes meet patients' psychological expectations, satisfy more The needs of multiple populations , our team is back with a full programme of An in-depth clinical interview was conducted at with a number of Patients were surveyed on their concerns about the new drug and we wanted to get the public's honest opinion of the team's project.

Contribution

In communicating with patients , we learnt that When faced with a drug, their main concern is the exact efficacy and safety of the drug , followed by the side effects and financial burden brought by . At the same time, patients still have a conservative attitude towards bacterial therapies. However, the majority of patients expressed interest in the issues that our project aims to address and are looking forward to the future.

Implementation

We respected and valued the privacy of our patients, and after signing an informed consent form for the interview under the condition that it was completely voluntary, we talked to several patients for about ten minutes each. (We strictly controlled the time and number of patients to ensure that we were as patient-centred as possible and to prevent them from feeling uncomfortable or offended.) In addition, we created and printed out a table of issues that might be considered when administering medication, and invited patients to write down the order in which they considered them. After statistical analysis, we found the issues that mattered most to patients based on their chemotherapy drugs - efficacy and safety topped the list, with concerns about side effects and financial pressure not far behind. We are very pleased to see that our project is "targeting" these patients' most important concerns.

Outlook

In presenting our programme to patients, we find that the general public still holds certain wariness and trepidation about the idea of bacterial therapy. "Pumping my body with bacteria that are already making people sick?" We know that there is still a long way to go to correct future perceptions about bacteria and even synthetic biology. Therefore, we hope to introduce synthetic biology to the general public in a more accessible and lively way, and to work towards changing the public's perception of emerging medicines.

Objective

In a conversation with one of our patients, we asked if he would be willing to try our bacterial therapy. Among some of the more resistant responses, one patient answered, "I trust my doctor, and I will use it as soon as Dr Wang tells me to." We were curious to see how Dr Wang got his patients to trust him so much, and we would also like to hear from radiotherapists about what they think of our project, and whether it can provide food for thought and inspiration from a different perspective than chemotherapy.

Contribution

Although a radiologist, Mr Wang expressed positive optimism about the prospect of biotherapies, suggesting that the issue of drug resistance can even be extended to the field of reducing radiation resistance, giving radiotherapy a certain synergistic effect. In addition, the teacher highlighted the issue of humanistic care in the whole process of cancer diagnosis and treatment. He advocates all-round care for patients' physical, mental and social attributes, from nutrition to psychological counselling, and he pays attention to all the details that promote patients' recovery.

Implementation

As this interview may involve the patient's medical history or private information, we drew up and informed Mr Wang to sign an informed consent form. Teacher Wang believed that the future of cancer treatment may lie in biotherapy, tumour vaccines and other precision treatment modalities, and affirmed our idea and project. In addition, the teacher told us that under the medical level of today's society, although it is possible to prolong the survival cycle of cancer patients to a greater extent, the physical and mental pain that cancer patients have to bear when undergoing treatment is enormous, and at the same time, every cancer patient will bring great financial pressure to his or her own family, and if our project can be steadily applied to the clinic, it will bring a certain degree of improvement to these problems. If our project can be steadily applied in clinic, it will bring some degree of improvement to these problems. In addition, the teacher's high medical ethics and meticulous care for the patients are also affecting everyone. The trust that patients place in their lives comes from the empathetic dedication of doctors. "There are times when the most important thing in surviving cancer is that will. And while treatment is only a small part of the disease process, the long recovery period and possible recurrence are also tough challenges. The patient's mind and body have to recover, especially the heart. Humans are social creatures, and our ultimate goal is for the patient to achieve healing in terms of social attributes."

Outlook

After hearing about the current situation, our whole team's heart is undoubtedly heavy. We deeply sympathise with the situation of each and every oncology patient, but it also affirms our direction and motivates our team. Next, we will continue to improve our project and conduct simultaneous analyses in both entrepreneurial and ethical threads to realise its clinical value as soon as possible. In addition, the holistic care and humanistic care proposed by Mr Wang has also aroused our thinking on social ethics. We will carry through the concept of human-centredness in the implementation of our future projects, raising the expected effect of the drugs developed by the team from treatment to cure, and paying attention to and valuing the psychological health and social role transformation of the patients, after all-"People ignore design that ignores people".

Objective

As the project advances to its final stages, the most significant challenge faced by the JLU-NBBMS team is how to coherently summarize and ensure a smooth transition of our overall design into the practical application phase. On September 8, 2024, we had the honor of inviting Director Dr. Cui Jiuwei from the Oncology Department of the First Clinical Medical College of Jilin University for an online interview. We hope to gain in-depth insights and suggestions from the expert to help the team have a clearer grasp of experimental details and a more coherent and comprehensive understanding of the project's overall design. Aiming to return to the modular core concept of synthetic biology, we leverage the advantages of modular design to systematically organize and summarize the project, ensuring that it becomes a safe and effective anti-tumor treatment system that can reduce drug resistance, precisely deliver medication, and minimize side effects.

Contribution

Through the exchange with Director Dr. Cui Jiuwei, the team was able to integrate the expert's clinical experience and scientific knowledge into the project design. This not only enhanced the scientific rigor and practicality of the project but also brought new perspectives and potential therapeutic strategies to the field of oncology treatment. Furthermore, by employing a modular design approach, we systematically optimized the project. This method aids in improving the project's flexibility and scalability, enabling it to better adapt to diverse therapeutic requirements and environmental changes.

Implementation

Synthetic biology emphasizes the decomposition of complex biological systems into standardized, controllable functional modules. This approach not only facilitates system controllability and optimization but also provides flexibility for future iterations and innovations. In our in-depth exchange with Director Cui, we recognized the importance of modular design. This design philosophy not only makes our project more organized but also enhances the system's controllability and scalability. Modular design allows us to independently test and optimize each component before integrating them into a larger system. The flexibility of this method means that we can easily replace, upgrade, or reconfigure modules in the future to accommodate new scientific discoveries or technological advancements.

The expertise and experience of Director Cui have had an immeasurable impact on our project. She not only patiently answered our technical questions regarding the safety of nanoparticles but also proposed innovative and feasible suggestions for our project. Her insights have helped us identify potential risks and areas for improvement within the project, allowing us to advance with greater confidence.

TUnder the guidance of Director Cui, we have re-evaluated our project and deconstructed it into four key modules: the Targeting module, the Gene regulation module, the Safety module, and the Drug delivery module. Each module undertakes specific functions and can be independently optimized and tested. We have analyzed the expanded functions of each module. [Learn more in Description]

Through the collaborative function of these four modules, our oncology treatment system is not only capable of significantly reducing tumor drug resistance, but also enhances the safety and efficacy of treatment by precisely delivering chemotherapeutic drugs. We believe that by continuously optimizing and expanding the functionality of these modules, our research will provide new ideas and methods for future cancer treatments. (Learn more in experiment)

Outlook

Through the exchange with Director Cui Jiuwei, our team has been able to refine our project and provide a new paradigm for the application of synthetic biology in the medical field. We also hope that our project will contribute to advancing research and applications of synthetic biology in oncology, offering valuable experience and references for future researchers and clinicians. This exchange has propelled the development of the JLU-NBBMS team's own project, delving into the modular design philosophy of synthetic biology. Moving forward, we will continue to seek new methods of improvement and present and exchange our project on more authoritative platforms.

Objective

The exchange with Director Cui has greatly helped the JLU-NBBMS team to refine the entire project. To date, we have modularized the project in line with the principles of synthetic biology. In order to better showcase our project, which is based on bacteria-iterative engineered biological targeting systems to assist drug delivery and enhance chemotherapy sensitivity, and also to further improve and refine, the JLU-NBBMS team participated in the National Virtual Teaching and Research Office for Biomedical Sciences Specialty 2024 Work Conference and the Biomedical Sciences Undergraduate Major Academic Exchange Meeting, held at the First Clinical Medical College of Jilin University on August 15, 2024. We hope that through this exchange, we can gain valuable opinions and suggestions from experts across the country, which are crucial for the further optimization and improvement of our project. We believe that through in-depth exchanges and discussions with peers, new ideas can be stimulated, potential problems can be identified, and more room for improvement can be explored.

Contribution

Through the exchange of research findings with leading academics, JLU-NBBMS has provided a novel perspective and direction for synthetic biology in the fields of reducing tumor drug resistance and mitigating the side effects of chemotherapy drugs. This has, to a certain extent, propelled research and practice in this area, with the hope of facilitating significant advancements in cancer treatment. Additionally, our participation has helped to disseminate knowledge related to synthetic biology nationwide, enhancing the public's understanding of this emerging scientific field. Lastly, with the critical guidance from attending experts, our project will be further enhanced, which is not only a valuable learning opportunity for our team but also contributes to the development of the biomedical sciences field. This exchange meeting has allowed us to establish connections with more peers, jointly promoting the application and advancement of synthetic biology in the field of biomedical sciences.

Implementation

The JLU-NBBMS team is consistently committed to pioneering new methods and possibilities for cancer treatment. On the eve of the exchange meeting, we engaged in in-depth communication with our mentors, aspiring to present a detailed and clear project at the conference. We began with an analysis of the current state of cancer, delving progressively into our project. From the project background, we identified the two major challenges faced in cancer treatment today—tumor drug resistance and the side effects of chemotherapy drugs. Having established our research direction, we addressed each issue methodically. In response to these two key problems, we elaborated on our project design, ensuring that every step of our design inspiration and thought process is captured by the experts and peers, culminating in a comprehensive experimental plan. This exchange meeting allowed us to establish connections with more colleagues, jointly promoting the application and advancement of synthetic biology in the field of biomedical sciences.

The International Genetically Engineered Machine Competition (iGEM) embodies the principles of synthetic biology; hence, we initiate from a modular design perspective, sharing our project more thoroughly with a different viewpoint. We have deconstructed the project design into smaller modules, each capable of independently performing its specific function. Moreover, due to the replaceability of the "parts," an iterative and personalized design is achieved, offering possibilities for treating various types of cancer. Each module also interacts with others, amplifying their effects.

Under the guidance of our expert mentors, we have conducted in-depth reflections and meticulous adjustments on various detailed aspects of the project. Meanwhile, the affirmation from our expert mentors has also filled the entire team with confidence, motivating us to transform this positive momentum into the success of the project and continuous innovation. We are well aware that the achievement of every project is inseparable from the joint efforts of the team and the dedicated instruction of experts. Therefore, we will continue to move forward hand in hand, constantly striving for excellence.

We have organized the overall structure and documentation of the project to ensure that all research steps, results, and conclusions are clear, logical, and easily comprehensible. Such organization not only aids in our own research work but also enables other researchers to more readily understand and utilize our findings.

Outlook

The 2024 National Virtual Teaching and Research Office for Biomedical Sciences Specialty Work Conference provided the JLU-NBBMS team with a more authoritative platform to showcase and exchange our project. Under the guidance of experts and teachers, our project has been further improved. Throughout the entire HP event, we delved deeper into the project, and these adjustments and optimizations have led our project to become more mature and refined. Moreover, the JLU-NBBMS team continues to popularize knowledge related to synthetic biology and the current state of cancer. We also look forward to our project providing new ideas and directions for cancer treatment, making a greater contribution to research in the fields of drug delivery and enhancement of chemotherapy sensitivity within synthetic biology. We look forward to joining hands with everyone to promote the advancement of science. This exchange meeting allowed us to establish connections with more peers, jointly promoting the application and development of synthetic biology in the field of biomedical sciences.

Introduction

As an undergraduate team from the Basic Medical College of Jilin University, JLU-NBBMS is dedicated to exploring innovative reforms that integrate medicine, engineering, humanities, and sciences, as well as fostering the development of foundational and applied research in the healthcare industry.

Purpose

Our objective during the pitching sessions of the 2024 (The 6th) World Health Expo was to present our project to various companies and to gauge the clarity of our design, concept, and market relevance.

Contribution

We visited multiple companies' booths, learning about the unique research and development projects and strategies employed by different hospitals. Throughout our visit, we engaged in conversations with various firms. We spent three minutes presenting our project and received feedback from ix biopharma. We discussed the advantages and disadvantages of different drug delivery systems and learned about the varying costs and production pipelines associated with each.

Initially, we focused on modeling simulations using several mature delivery systems available in the market. Through ix biopharma's staff, we communicated with their managers. We also realized that, at our current stage, enterprises focusing on a single link are unlikely to take interest or collaborate with us. For a vast pharmaceutical production system, we need to instill sufficient confidence in investors to facilitate cooperation.

Implementation

Exploring different drug delivery methods and conducting a cost analysis will help us propel our project to a new phase. Therefore, we also described our ideas more effectively in subsequent events and conversations with stakeholders. Based on our discussion with ix biopharma, we formulated preliminary ideas for our model section: to simulate the efficiency of different delivery systems to comprehensively evaluate the final experiments, starting from freeze-dried powder.

Outlook

This iterative process of pitching and refining is a crucial characteristic of disseminating our ideas to society and various stakeholders and investors. We hope that each conversation brings us one step closer to the commercialization of our project.

Introduction

After gaining insights into the biopharmaceutical market in China and the development of domestic enterprises, we have advanced our design to incorporate common chemotherapy drugs. As one of the most advanced biopharmaceutical companies in the region, we aspire to visit mature biopharmaceutical industries to understand their entrepreneurial and developmental journey, as well as their efforts in nanotechnology.

Purpose

Our aim is to communicate with pharmaceutical companies, exchange ideas with industry practitioners among our stakeholders, and learn how to conduct industry and competitive market analysis. Additionally, our goal is to comprehensively understand the supply chain and development process of cell therapies. Jin Lei has extensive experience in developing cell therapy drugs. RNAi therapy is a treatment for various cancers that silences tumor-related genes in patients to control and kill tumors. Our team envisions integrating this therapy with existing chemotherapy.

Contribution

The most significant feedback we received from pharmaceutical companies regarding the overall planning of BIOTARGET is the focus on post-commercialization cost control. Our idea of using nanoparticles to encapsulate different chemotherapy drugs for a full solid tumor treatment plan was also rejected. During communication with gensci, Ma Ji, an expert in marketing, explained in detail that if we initially plan our product to cover too broad a range, the price and production costs would increase exponentially, making the treatment unaffordable for the market. Therefore, after understanding the current state of the industry, we should engage in in-depth communication with patients to understand the size of the patient group, the dosage each patient requires, and their acceptance of different treatment plans, thus further adjusting our overall planning and experimental validation stages. Gensci believes that BIOTARGET will bring new vitality to the market because it is an evolving treatment technology that learns from the best. However, Jin Lei advises us to consider the safety of the bacterial strains we use in the approval process and when treating patients. He also suggests that we should communicate and exchange ideas with more enterprises in other regions. Additionally, Ma Ji provided feedback on the commercialization of scientific research projects and the transformation of school-enterprise projects, expressing his hope that we can develop a reliable plan through continuous design and improvement to gain market recognition.

Implementation

Based on the feedback on our cost structure, we have conducted multiple rounds of surveys targeting different income groups and individuals with varying levels of knowledge. Additionally, we calculated and discussed the size of the solid tumor patient population we could reach. The human practice team discussed which experimental experts and market-oriented transitions are needed to more accurately confirm the transformation of our experimental projects. Also, we discussed which experts could teach us how to scale out BIOTARGET.

Outlook

To further investigate the design of our cell therapy and corresponding safety measures, our goal is to interview experts in nanomaterials and tumor treatment. Specifically, our aim is to speak with Wang Fang, Dean of the Translational Medical Research Institute at Jilin University, and Xu Caina, an expert in nanomaterials at the Basic Medical College of Jilin University. Furthermore, we plan to collaborate with experts related to nanomaterial science at the School of Chemistry at Jilin University. Our goal is to use these interviews to validate the information we obtained from gensci, concluding the feedback cycle.

Introduction

The Shenzhen Industrial Innovation Center for Engineering Biology is an industry hub dedicated to fostering innovation and entrepreneurship in synthetic biology. To understand the feasibility of our project design from the perspectives of university medical translational research institutes and pharmaceutical companies, we were invited by teams such as BNUZH-China to participate in the second iGBA Exchange Forum. During the forum, we visited the Shenzhen Industrial Innovation Center for Engineering Biology, where we had discussions with the person in charge, Dang Xiaomeng, to gain insights and advice on the industrial implementation of BIOTARGET.

Contribution

Dang Xiaomeng showed great interest in our project and its innovative nature, introducing us to the facilities and policy support available at the center. However, there were concerns about the feasibility of bringing this therapy to market. They mentioned that bacteria-based therapies are not without risks. For instance, they could cause side effects or provoke immune responses against the injected Salmonella or nanoparticles.

The use of Salmonella in humans might trigger an immune response. Even if we sufficiently attenuate the toxicity of Salmonella, we still need to ensure that it is recognized by the immune system to induce the body's immune response to target the tumor microenvironment along with Salmonella. Additionally, there is hesitancy regarding the effectiveness of Salmonella's environment-specific targeting. Concerns arise whether Salmonella will aggregate at the intended site due to similar conditions present in other environments, and how long it would take for Salmonella circulating throughout the body via the secretion system to lyse and die in other areas to ensure safety.

Implementation

The feedback on potential immune and toxicity reactions caused by the use of Salmonella and nanoparticles is an area we have been contemplating and improving. This has prompted us to consider which cells should be engineered and implemented in our BIOTARGET. Furthermore, due to the risks associated with bacterial therapy, we are urged to conduct a strict review of bacterial safety and to thoroughly read the policies in the relevant review areas. Du Wenjin, another colleague at the center, also expressed concerns about the safety and efficacy of our engineered bacteria. We have rigorously reflected on this part of our engineered bacteria using findings from literature and feedback from expert meetings.

Outlook

The next step is to decide which factors we need to consider when scaling up the experimental content involved in our experiments to an industrial pipeline. Additionally, we will discuss with stakeholders (clinical physicians and clinical trial doctors) the types of tumors we aim to address.

Introduction

Shenzhen Synthetic Biology Infrastructure is an industry transformation center for synthetic biology, supported by schools, research institutes, and the government. To further understand the support situation of the synthetic biology industry in different regions and the feasibility of college student entrepreneurship, I attended the second iGBA forum and visited Biosysen and Biocretech companies, meeting with the founder of Biosysen, Liu Yujia. Liu Yujia provided us with advice on the support policies in Guangming District, Shenzhen, and the feasibility of college student entrepreneurship.

Contribution

The support policies for synthetic biology in Guangming District, Shenzhen, are very advanced. Here, companies and enterprises of different categories can share a set of laboratory equipment, thereby reducing the financial and procurement pressure on startups. At the same time, different companies can help each other with their products. For example, when Liu Yujia introduced the company and the situation of related research centers, thanks to the industrial cluster and cost advantages, the laboratory materials produced by Biosysen can be supplied to surrounding companies and have gradually become a part of Biosysen's revenue. This also provides us with a new idea: if we can land our project in a base with a complete surrounding industrial chain in the future, we can cooperate with surrounding companies to achieve an "internal circulation" within the industrial park, operating in multiple fields while minimizing costs, and we can transform our company's business strategy into a model that focuses on the development of new drugs, supplemented by the supply of experimental by-products.

At the same time, we learned about a new form of cooperation between industry, schools, and research institutes at Shen Zhen Synthetic Biology Infrastructure, that is, research institutes and governments cooperate to run schools, and students are jointly cultivated by research institutes and companies in the industrial cluster, helping students to quickly transition from being school students to scientific research talents in companies. This has also inspired our subsequent work, shifting the focus from the transformation of a single project to the cultivation of a talent team.

Implementation

We have gradually understood the business models of enterprises in different regions of the country and the various considerations for college students starting businesses. In addition, through exchanges with different companies, we have also learned that there is a very important part in the future landing process of BIOTARGET - the two major links of drug approval and intellectual property protection.

Therefore, we continue to think about how to communicate with startups and sponsors in different regions to understand these two extremely important sections. In addition, we have also started to write our company's business strategy and look forward to providing enough information to the other party in subsequent exchanges.

Outlook

The next step is that we will decide on our company's operation mode and understand the approval processes for drug approval and the operation of biopharmaceutical enterprises. We choose to go to the UK to meet with the founder of a very successful startup, oxartis, at the Heyford Park Innovation Center, which has a similar model.

Introduction

Heyford Park Innovation Center, located just a few miles north of Oxford, UK, has developed over the years into a very successful small and micro-enterprise startup center in Oxford. Similar to the facilities center in Shenzhen, it can alleviate the pressure brought by purchasing equipment and also provide some financial support. We met with Dr. Julian Dye, the founder of one of the outstanding startups, oxartis. He introduced us to the journey and considerations of being a startup founder.

Contribution

Dr. Julian Dye introduced us to their six-person startup, oxartis, a biomaterials company focused on bio-tissue regeneration scaffolds. Their company targets the clinically unmet demand for scalable regenerative biomaterials and has pioneered a new type of vascularized dermal scaffold for skin injury reconstruction.

In their introduction, Dr. Julian Dye mentioned, "If you want your research to become a product, you must treat it with a commercial mindset from the very beginning." This was very enlightening to me. Secondly, they conducted research and surveys on potential users and also understood the regulatory environment and requirements. At the initial stage, they used digital simulation to simulate the final results of their experiments, which were then compiled into materials and submitted, successfully obtaining the Puffin Innovation Fund, thus starting the product development work. In addition, they also provided us with a lot of experience in market analysis and intellectual property protection. Especially in the aspect of intellectual property protection, it broke my previous understanding that intellectual property protection requires a series of protective measures like a dense net to ensure product benefits.

At the same time, he also mentioned something very interesting. When applying for intellectual property rights, we need to apply for the core technology patents without revealing the core technology, which is a very contradictory thing. You cannot guarantee that those who review these materials will not reverse develop your patents or even get ahead of you. At the same time, operating freely is also a very special part. We need to understand that our company and other companies do not completely overlap, which will help you not be suppressed for a period of time. Therefore, you must make your project unique in order to stand out among competitors.

Implementation

Under the introduction of the professor, we have gradually optimized our overall commercial route and continue to come up with new ideas in seeking potential investigations of doctors and potential needs of patients. The road to commercialization is very long. We have set a long-term schedule and hope to continue to optimize the entire project and truly achieve industrialization after the iGEM competition. In addition, the research we have done on stakeholders should not be enough, so next, we will continue to screen stakeholders and conduct comprehensive interviews and research.

Outlook

The next step is for us to further understand the content of intellectual property application and approval processes. The content we need to consider is very complex and has come to a new field. For this reason, we went to the Changchun Intellectual Property Protection Center and communicated online with Beijing Jingjinshi Intellectual Property Agency Co., Ltd.

Introduction

Beijing Jingjinshi Intellectual Property Agency Co., Ltd. is a well-established intellectual property application company. We seek such a company to provide us with preliminary intellectual property analysis and a network of intellectual property protection for our BIOTARGET. Additionally, such a company is an essential part of our process of transforming from the laboratory to entrepreneurship. We wish to understand the key points to consider when a company applies for intellectual property patents. Hope to understand the relevant policies and laws and regulations for intellectual property protection at the government level, we also visited the Changchun Intellectual Property Protection Center.

Contribution

Liu Guanyu, a representative from Beijing Jingjinshi Intellectual Property Agency Co., Ltd., provided us with valuable insights into the intellectual property landscape. They emphasized the importance of early and strategic patent filing to secure a strong position in the market. Liu Guanyu highlighted that understanding the patentability criteria, conducting thorough prior art searches, and crafting a well-structured patent application are crucial steps in the patenting process.

Liu Guanyu also pointed out that we should consider the international protection of our intellectual property, especially if we plan to expand globally. They advised us on the different routes available for international patent applications, such as the Patent Cooperation Treaty (PCT) system.

Moreover, Liu Guanyu discussed the significance of non-disclosure agreements (NDAs) and material transfer agreements (MTAs) in protecting our proprietary technology during collaborations or when engaging with third parties.

Implementation

Based on the guidance from Beijing Jingjinshi Intellectual Property Agency Co., Ltd., we have initiated the process of patent drafting and filing. We have engaged in a series of brainstorming sessions to identify the innovative aspects of our BIOTARGET that can be protected by patents. We are also in the process of conducting a comprehensive prior art search to ensure that our inventions are novel and non-obvious.

We are developing a strategy to manage our intellectual property portfolio, which includes deciding which markets to target for patent protection and setting a timeline for filing applications. Additionally, we are creating a protocol for documenting our research and development process to support our patent claims.

Furthermore, we are engaging with government agencies to understand the local and national policies that could support our intellectual property protection efforts. We are also exploring potential funding opportunities and incentives for startups focusing on innovation and intellectual property.

Outlook

Looking ahead, our next steps involve finalizing our patent applications and filing them with the appropriate authorities. We will continue to work closely with Beijing Jingjinshi Intellectual Property Agency Co., Ltd. to navigate the complexities of the patent system.

We are also planning to engage with potential industry partners to discuss licensing opportunities and collaborative research. This will help us assess the market potential of our BIOTARGET and identify any additional intellectual property that we may need to protect.

Finally, we will keep abreast of any changes in intellectual property laws and regulations, both domestically and internationally, to ensure that our strategy remains compliant and effective. Our goal is to establish a robust intellectual property protection network that will support the commercialization of our BIOTARGET and secure our competitive edge in the market.

Introduction

The iGEM StartUp community has been a focal point for JLU-NBBMS since our team's inception. We have been closely following the blogs and events they publish. After conducting extensive research and forming a preliminary business plan, we took advantage of the iGEM StartUp Summer School to engage in online exchanges and discussions with iGEM teams from around the world who are also interested in entrepreneurship, hoping to draw inspiration from different teams.

Contribution

Following the opening remarks by the director of iGEM StartUp and the director of iGEM Judging, we were continuously placed into different groups to introduce our projects. In the process of exchanging with other teams, we were delighted to learn about the research content of teams from Greece, Pakistan, France, Mexico, and other countries, and to understand the general situation of medical policies in different regions. This enables us to better achieve "when in Rome, do as the Romans do" in the localization of our products in different areas.

Implementation

Through this Summer School, our team continuously established connections with other teams and tried to understand their projects as much as possible, working with them to improve the entrepreneurial aspects of our projects and achieve overall improvement. Under the guidance of the director, we also learned how to conduct and practice the entrepreneurial part within the iGEM community and how to raise funds from iGEM StartUp, thereby truly starting from iGEM and promoting iGEM.

Outlook

1. Collaborate with International Teams: Establish partnerships with teams from different countries to explore potential collaborations and exchange of knowledge.

2. Localize Our Approach: Tailor our product development and marketing strategies to fit the specific needs and regulations of various regions.

3. Refine Our Business Plan: Incorporate the feedback and ideas from the Summer School to enhance our business plan.

4. Explore Funding Opportunities: Investigate the funding opportunities provided by iGEM StartUp and other sources to support our entrepreneurial endeavors.

5. Develop a Marketing Strategy: Create a comprehensive marketing strategy that leverages the iGEM brand and community to promote our project.

6. Prepare for iGEM Competition: Use the knowledge and connections gained to strengthen our iGEM competition entry and maximize our chances of success.

7. Long-Term Entrepreneurial Goals: Set clear goals for taking our project beyond the iGEM competition and into the startup phase, including planning for product development, market entry, and scaling up operations.

By following these steps, we aim to transform our iGEM project into a successful startup that not only wins in the competition but also makes a significant impact in the real world.

Introduction

After gaining a preliminary understanding of the market, market analysis and the analysis of competing enterprises are well underway. A critical step that follows is Quality Manage System (QMS), which is integral throughout the entire translational process. From the beginning of approval to small sample trials and adjustments, and finally to the establishment of a production pipeline, every step involves QMS. To gain insights into how quality control is approved at various levels and is closely related to project translation and production, we visited Changchun LanJiang Pharmaceutical Technology Co., Ltd. Jiang Bo, the founder of the company, answered all our questions, and after Dr. Shang, the Operations Manager, led us through the workshop and shared some quality control documents with us, we gained a basic understanding and are ready to continue refining our business plan.

Purpose

Our aim is to communicate with pharmaceutical companies, exchange ideas with industry practitioners among our stakeholders, and learn how to conduct industry and competitive market analysis. Additionally, our goal is to comprehensively understand the supply chain and development process of cell therapies. Jin Lei has extensive experience in developing cell therapy drugs. RNAi therapy is a treatment for various cancers that silences tumor-related genes in patients to control and kill tumors. Our team envisions integrating this therapy with existing chemotherapy.

Contribution

Following our visit to Changchun LanJiang Pharmaceutical Technology Co., Ltd, we gained profound insights into the pivotal role that Quality Management System (QMS) plays throughout the product development and commercialization process. Jiang Bo, the founder of the company, elucidated how QMS begins from the conceptual stage of a project and continues through to product approval and production. She emphasized that quality control is not just about regulatory compliance but is also the cornerstone of a company's sustained success and product reliability.

Dr. Shang, the Operations Manager, provided us with a practical understanding of the day-to-day operations of QMS by giving us a tour of the manufacturing facilities and sharing quality control documents. We learned how to establish and implement Standard Operating Procedures (SOPs), conduct quality audits, and ensure quality through continuous process validation and product testing.

This experience was invaluable as it helped us understand not only the importance of QMS but also the practical details of implementing QMS during product development.

Implementation

Based on the knowledge acquired at Changchun LanJiang Pharmaceutical Technology Co., Ltd, we began to refine our own QMS. We started by evaluating our existing operational processes and identifying areas for improvement. We are developing a set of SOPs to ensure that every step, from laboratory research to production, is guided and documented clearly.

We are also designing an effective quality audit system to regularly inspect our processes and products, ensuring they meet established quality standards. In addition, we are working with quality control experts to ensure that our QMS meets all applicable regulatory requirements.

To integrate QMS into our product development cycle, we are establishing a cross-functional team, including representatives from R&D, production, quality assurance, and regulatory affairs. This team will be responsible for overseeing the implementation of QMS and ensuring that all relevant personnel receive proper training.

Outlook

Looking ahead, we recognize that the implementation of QMS is an ongoing process that requires continuous evaluation and improvement. We plan to regularly review our QMS to ensure it adapts to new challenges and changes in the regulatory environment

We will also explore how to utilize advanced quality control technologies, such as automated testing and data analysis, to improve our efficiency and accuracy. Furthermore, we are considering establishing a feedback mechanism to gain valuable insights from customers and the market so that we can continuously improve our products and processes.

Ultimately, our goal is to establish a QMS that meets regulatory requirements and supports our business growth. By doing so, we believe we can deliver high-quality products that satisfy market demands and create value for our investors and customers.

Introduction

Dirui Medical Technology is an experienced diagnostic and testing enterprise, specializing in tracking and monitoring disease markers, especially oncological markers. We aim to meet with Dirui to understand how a company with extensive experience can ensure profitability while screening a multitude of tumor markers, or in other words, how to determine that these tumor markers are needed by the market.

Contribution

Guided by the person in charge at Dirui, we conducted a tour and interview. From their description, we gradually understood the history of how the company has grown and developed over time. When it comes to monitoring various markers, we learned how they can maintain a considerable revenue while monitoring such a wide range of categories. The person in charge told us that it's not just about the size we achieve at present to speculate our capabilities; Dirui started with a few commonly seen and necessary markers for approval processes abroad, gradually expanding to cover multiple categories and diseases to expand the market and improve the quality of the production line. This is a positive cyclical cultivation model, which requires the founder's leader to have a forward-looking vision and the ability to predict market trends in advance.

Implementation

Although Dirui is not engaged in the production and development of drug products, the key information they provide is also very valuable. We have always had certain misconceptions about the testing industry and the transformation of medical projects. We would stubbornly believe that testing institutions are just an amplification of our laboratory testing, without bringing a large product into the approval process, which requires multiple rounds of testing and inspection to ensure the quality of each batch of products.

Outlook

The next step is to explore the content that needs to be considered throughout the entire process of product research and development and transformation, which is the Quality Management System (QMS). This is very important, as it relates to our product achieving a standard therapeutic effect during mass production. We will delve into understanding the QMS framework, which includes aspects such as:

1. Quality Planning: Defining the quality standards and objectives for our product.

2. Quality Control: Implementing processes to monitor and control product quality during production.

3. Quality Assurance: Ensuring that our production processes and products meet the required quality standards.

4. Quality Improvement: Continuously reviewing and improving our QMS to enhance product quality.

5. Regulatory Compliance: Making sure that our QMS complies with all relevant regulations and guidelines.

6. Risk Management: Identifying potential risks and implementing measures to mitigate them.

By focusing on these aspects, we aim to establish a robust QMS that will guide us through the product development process, ensuring that our BIOTARGET not only meets the market's needs but also maintains a high standard of quality and safety.

Introduction

After conducting a series of research and modifications, we began to gradually build our own integrated platform to manage the entire team. Having used many common office software applications, we often encountered issues with usage, including costs paid to the companies and the need to separate our personal software from our work software. Therefore, we are also continuously searching for a platform suitable for experimental developers and optimized for bacterial editing, capable of realizing project management, product design, and the final output of cloud-based samples. Thus, we collaborated with Yanyin Technology to address the first half of the pain points and with Shutong Technology to solve the latter half of the issues encountered.

Contribution

Yanyin Technology is an innovative technology company driven by biomedical AI big models, committed to creating a leading biomedical digital intelligent scientific research collaboration platform in China, becoming a connector in the biomedical scientific research ecosystem, and promoting efficient, compliant, and intelligent life science research and development. We had a brief contact with Yanyin Technology during the CCiC and learned that their products meet our current needs, so we proceeded with three communications.

Dr. Chen Zeping, the chairman of Yanyin, highly praised our project and provided us with a new perspective from an empowerment enterprise angle. Every company and enterprise is continuously improving its overall blueprint to achieve ultimate victory; hence, we must understand how to cooperate and achieve a win-win situation.

Implementation

With Yanyin Technology, we are developing an online automated platform tailored to our team's specific needs. This platform will integrate project management, product design, and sample output functionalities, streamlining our workflow and ensuring that all team members have access to the most up-to-date information and resources.

In collaboration with Generulor, we are addressing the optimization and improvement of bacterial editing. This involves creating a platform that can manage and track the progress of bacterial modifications, ensuring that our research remains at the cutting edge of the field.

Outlook

Looking forward, we aim to have our online automated platform fully operational, enabling seamless collaboration and data management among our team members. We will continue to refine the platform based on user feedback and technological advancements.

We plan to conduct a series of training sessions to ensure that all team members are familiar with the platform and can utilize it to its full potential. Additionally, we will explore opportunities to integrate third-party tools and applications that can further enhance the platform's capabilities.

Purpose

In the process of translating last year's iGEM project in collaboration with gensci, we were advised to consider the entire drug development process when designing new therapies. To kickstart our business plan, we have had some interesting discussions with several local companies. However, we still have questions regarding our SWOT (Strengths, Weaknesses, Opportunities, and Threats) analysis and financial analysis. Bioxun specializes in the production and marketing of medical products. Therefore, we decided to schedule a meeting with gensci to get feedback on the feasibility of our business idea, SWOT analysis, and the operational model of our BIOTARGET.

Contribution

Regarding the SWOT analysis, we received some valuable insights. Dr. Liu, the company manager, stated that we must consider our weaknesses and threats. They emphasized that every weakness or threat hides opportunities. For instance, they explained that marketing products might be challenging for a small company. Therefore, leveraging the name of a large company could help expand your network and add credibility to your product. For financial analysis, they suggested breaking down the entire project into smaller phases such as R&D, clinical trials, manufacturing, legislation, marketing, etc., and identifying the estimated costs for each phase to determine the total cost.

Implementation

We expanded our SWOT analysis based on Dr. Liu's feedback, understanding that threats and weaknesses are not necessarily bad but should be seen as opportunities. For every weakness and threat, we are looking for new ways to mitigate them or use them to our advantage. We divided the project process into several smaller phases and attempted to forecast the costs separately for each. This greatly helped us in formulating a more realistic financial plan. However, due to the inherent uncertainties, we had to make many assumptions about the R&D phase, clinical trials, production, and legislation.

Outlook

To understand the complete drug development process and implement a more systematic approach to determining the costs at each stage, we believe it is valuable to talk to other stakeholders. In the next steps, our goal is to identify and engage with stakeholders relevant at each stage of the drug development pipeline. Since Bioxun is not fully clear, we also plan to contact Wei Jun again. As the person in charge of the earliest I-phase clinical trials, we believe he will provide us with advice on different perspectives of stakeholders. Finally, we must decide on a business strategy to address this issue; how will we market and sell BIOTARGET?

Introduction

Professor Wei Jun is the director of the Phase I Clinical Laboratory at the Third Hospital of Jilin University, also known as Baiqian Third Hospital. He is the first communicator and contact person who helps to promote products through clinical approval to the hospital. Professor Wang Tianwei is an expert in the field of oncology at the same hospital. We hope to learn about the latest trends in updated oncology treatment guidelines from our conversation with him.

Contribution

Our discussion with Professor Wei Jun provided us with a clear understanding of the clinical trial research approval process. His insights into navigating the regulatory landscape for clinical trials were invaluable. He emphasized the importance of rigorous testing and the necessity of adhering to ethical standards throughout the process.

From our conversation with Professor Wang Tianwei, we gained insights into the current oncology treatment guidelines. He shared the latest trends in cancer treatment, including new approaches, therapies, and the integration of personalized medicine. His expertise helped us understand how these guidelines could impact the development and potential approval of our BIOTARGET.

Implementation

With the knowledge gained from Professor Wei Jun, we are refining our approach to clinical trial design and patient recruitment. We are ensuring that our protocols are in line with the latest regulatory requirements and that they are ethically sound. We are also considering his advice on how to effectively communicate the benefits and risks of our BIOTARGET to potential participants.

Professor Wang Tianwei's guidance on the current state of oncology treatments has helped us align our BIOTARGET with the latest medical standards. We are incorporating his suggestions into our product development strategy to ensure that our therapy is not only innovative but also compatible with current clinical practices.

Outlook

Looking forward, we plan to maintain a close dialogue with both Professor Wei Jun and Professor Wang Tianwei. We aim to keep our clinical trial protocols updated with the latest regulatory changes and ensure that our BIOTARGET remains at the forefront of oncology treatment.

We will also explore potential collaborations with their departments for clinical trials and seek their advice on patient stratification and treatment personalization. By integrating their expertise, we hope to enhance the clinical relevance and acceptance of our BIOTARGET, ultimately improving patient outcomes.

In conclusion, our engagement with these key opinion leaders will be an ongoing process, ensuring that our product development is informed by the latest clinical insights and regulatory standards.

Introduction

OBIO is a high-tech company deeply engaged in the core fields of cell and gene therapy. They specialize in providing CRO services for basic research in cell and gene therapy, including the development of gene therapy vectors, gene function research, and drug target and efficacy studies. Additionally, they offer CDMO services for the research and development of cell and gene therapy drugs, such as process development and testing, IND-CMC pharmaceutical research, and GMP production of clinical samples and commercial products. Their scope of services also extends to CDMO services for the commercial production of bacterial therapy products. Therefore, we met with the person in charge of OBIO to understand the specifics of the services they provide and what comprehensive GMP production and CDMO services entail.

Contribution

The meeting with OBIO provided us with a detailed overview of their services, particularly focusing on the CDMO aspect for bacterial therapy products. Their expertise in the cell and gene therapy domain has been instrumental in shaping our understanding of the complexities involved in taking a product from the research phase to commercialization.

We learned about the importance of GMP production standards in ensuring the quality, safety, and efficacy of biological products. OBIO's experience in IND-CMC pharmaceutical research was particularly enlightening, as it highlighted the regulatory requirements and the need for stringent quality control measures throughout the drug development process.

We expanded our SWOT analysis based on Dr. Liu's feedback, understanding that threats and weaknesses are not necessarily bad but should be seen as opportunities. For every weakness and threat, we are looking for new ways to mitigate them or use them to our advantage. We divided the project process into several smaller phases and attempted to forecast the costs separately for each. This greatly helped us in formulating a more realistic financial plan. However, due to the inherent uncertainties, we had to make many assumptions about the R&D phase, clinical trials, production, and legislation.

Implementation

Following our meeting with OBIO, we are now better equipped to plan the next steps for our product's development. We will be focusing on the following action items:

1. GMP Compliance: We will ensure that our production processes are designed to meet GMP standards to guarantee product quality and consistency.

2. IND-CMC Strategy: We will develop a robust IND-CMC strategy to navigate the regulatory pathway for our bacterial therapy product.

3. Partnership Exploration: We will explore potential partnerships with OBIO for process development, testing, and GMP production to leverage their expertise and facilities.

4. Quality Manage Systems: We will establish comprehensive quality control systems to monitor and ensure the efficacy and safety of our product at every stage of production.

5. Regulatory Engagement: We will engage with regulatory authorities to understand the specific requirements for bacterial therapy products and how OBIO can assist us in meeting those requirements.

Outlook

Moving forward, our goal is to establish a solid foundation for the commercialization of our bacterial therapy product. We will continue to engage with OBIO and other service providers to ensure that we have a comprehensive understanding of the entire production process.

We will also focus on building a strong relationship with regulatory agencies to ensure a smooth IND filing and approval process. Furthermore, we will keep abreast of the latest advancements in cell and gene therapy to incorporate the most innovative approaches into our product development.

In the long term, we aim to create a strategic partnership with OBIO that not only supports our current product but also positions us for future growth in the cell and gene therapy space. By leveraging their CDMO services, we can accelerate our path to market and improve our chances of success in the competitive biopharmaceutical industry.

Introduction

Jilin Weilai Kangyuan Pharmaceutical Technology Co., Ltd. is a highly successful example of a commercial project transformed from the Basic Medical College of Jilin University. We chose to meet with the company's chairman, Li Jing. We briefly described our own project and discussed with her the gap between research projects led by university professors and those of real scientific research companies. Li Jing believes that we should go out more, observe, and learn extensively. She thinks that the pace of company development has far surpassed the research conducted by university professors. The difference in motivation brought about by research purposes versus profit purposes is inestimable and shows an increasing disparity. Therefore, we hope to gain some insights from Dr. Li to help us change our thinking and solve problems.

Contribution

Our conversation with Dr. Li Jing provided us with valuable insights into the differences between academic research and industrial projects. She emphasized the importance of understanding the market dynamics and the need for flexibility and speed in a commercial environment, which can be quite different from the more methodical pace of academic research.

Dr. Li Jing shared her experiences in transforming scientific findings into marketable products and the challenges she faced in doing so. She stressed the importance of having a clear vision of the end goal and being able to adapt to changing circumstances. Her advice on how to bridge the gap between academia and industry was particularly enlightening.

Implementation

Inspired by our meeting with Dr. Li Jing, we are taking the following steps to align our project with commercial realities:

1. Market Research: We are conducting thorough market research to understand customer needs and identify potential competitors.

2. Business Model Development: We are developing a business model that not only focuses on scientific innovation but also considers profitability and marketability.

3. Business Model Development: We are developing a business model that not only focuses on scientific innovation but also considers profitability and marketability.

4. Adaptability: We are building a project team that is capable of adapting to changes in the market and regulatory environment.

5. Commercial Mindset: We are fostering a mindset that prioritizes the commercial viability of our project, in addition to its scientific merits.

Outlook

Looking ahead, we plan to continue learning from successful commercial projects like Jilin Weilai Kangyuan Pharmaceutical Technology Co., Ltd. We aim to integrate the lessons learned into our project development process.

We will also seek opportunities to collaborate with industry partners who can provide guidance and resources to help us navigate the commercialization process. Our goal is to create a product that not only advances scientific knowledge but also meets market demands and achieves commercial success.

Introduction

At the iGC forum, I focused on the drug approval segment, which involves both Chinese and American approvals. During my conversations with experts at the event, we continuously explored the differences in approval regulations and processes across various regions. The approval processes for China, Europe, and the United States all require in-depth understanding. Therefore, while attending the lectures, we integrated the regulations of each country with those of our own, forming a comprehensive set of policy explanations and analyses.

Contribution

The forum provided a platform to delve into the specifics of FDA IND submissions, particularly the Chemistry, Manufacturing, and Controls (CMC) section, which is critical for drug approval. The insights gained from experts have been instrumental in understanding the challenges and key points of IND submissions.

We learned that a robust CMC section is essential for a successful IND application. It requires detailed information on the drug's manufacturing process, its characterization, and the controls used to ensure the quality, purity, and strength of the drug. The experts emphasized the importance of early and continuous communication with regulatory agencies to address any potential issues proactively.

Implementation

Armed with the knowledge from the forum, we are taking the following steps to tackle the CMC challenges and prepare for our IND submission:

1. CMC Documentation: We are meticulously preparing our CMC documentation, ensuring that all aspects of the manufacturing process, from the drug substance to the drug product, are well-documented and cGMP standards.

2. Quality Control: We are establishing stringent quality control measures to ensure batch consistency and to provide data that demonstrates our drug's quality throughout its shelf life.

3. Regulatory Strategy: We are developing a regulatory strategy that includes a plan for ongoing communication with the FDA and other regulatory bodies to facilitate a smooth review process.

4. Pre-IND Consultation: We are planning to have a pre-IND consultation with the FDA to discuss our CMC data and address any concerns before submission.

5. Manufacturing Readiness: We are working closely with our contract manufacturers to ensure that they are prepared to scale up production in compliance with GMP regulations once our IND is approved.

Outlook

Looking forward, we understand that the process of IND submission is just the beginning of a long journey towards commercialization. We are committed to maintaining a high standard of quality and compliance throughout this process.

We will continue to monitor changes in regulatory guidelines and adapt our strategies accordingly. Additionally, we plan to engage with regulatory consultants and experts to ensure that our submission is as strong as possible.

In the long term, we aim to establish a solid foundation for our drug's approval and commercial success. By proactively addressing the CMC challenges and maintaining open lines of communication with regulatory agencies, we believe we can navigate the complex landscape of drug approval and bring our product to market efficiently.