The Public Survey

A detailed survey was conducted to evaluate the public’s understanding of pancreatic cancer and its treatment procedures. This study aimed to assess the existing level of knowledge regarding pancreatic cancer, its risk factors, and current treatment options, as well as to identify misconceptions and information gaps. These data helped us make informed decisions regarding the development of initiatives tailored to enhance public awareness and acceptance of advanced therapeutic strategies, such as dual-targeting nanobody.

Demographic Breakdown

The survey received responses from a diverse group of participants. Nearly 48% fell within the "18-34" age range, followed by the "35-54" category with 47% participants. A smaller number of respondents were under 18 or over 55 years old, allowing us to gain insights across various age groups.

Key Findings

Awareness of Pancreatic Cancer

The survey revealed a commendable level of awareness among participants, with 94% having heard of pancreatic cancer. This suggests that the general public has a baseline understanding of the disease, which is crucial for acceptance of new treatment modalities.

Perceived Commonality of Pancreatic Cancer

Despite the awareness, most respondents perceived pancreatic cancer as less common compared to other cancers, highlighting the urgency and support for research and development of novel treatments, underscoring the need to showcase the severity and rising incidence of pancreatic cancer in public discussions.

Awareness of Known Risk Factors

Most respondents demonstrated a high level of knowledge regarding risk factors, with more than 76% correctly identifying smoking, a high-fat diet, and family history as key contributors to the disease. However, nearly 24% of the participants were unsure, indicating that further education is necessary to reinforce awareness of these risk factors.

Knowledge of Common Treatments

Awareness of the multifaceted treatment approach for pancreatic cancer was high, with 70% of participants acknowledging the use of surgery, radiation therapy, chemotherapy, and targeted therapy. The selection of "All of the above" as the most common response reflects an understanding of the complexity of pancreatic cancer treatment.

Awareness of Targeted Drug Therapy

The survey revealed a very high level of awareness regarding targeted drug therapy among respondents, which is promising for the introduction of advanced treatments, such as dual-targeting nanobodies. This indicates readiness within the public to embrace novel therapeutic strategies that go beyond traditional chemotherapy.

Perceived Effectiveness of Targeted Drug Therapies

More than 50% of respondents believed that targeted drug therapies were more effective than traditional chemotherapy, showing a favorable perception; however, nearly 30% of respondents were either unsure or disagreed, reflecting a degree of skepticism that must be addressed through clear communication about the advantages and limitations of targeted therapies.

Conclusion

The survey data highlight both the strengths and gaps in public knowledge of pancreatic cancer and its treatment options. The widespread awareness of targeted therapies suggests an openness to innovative treatments such as dual-targeting nanobody. However, misconceptions about the prevalence of pancreatic cancer and uncertainty about the specific diagnostic and treatment methods signal the need for targeted educational initiatives.

Our strategy focuses on leveraging this foundational knowledge by providing clear, evidence-based information on the benefits and mechanisms of dual-targeting nanobodies, through educational campaigns designed to emphasize the effects of pancreatic cancer and the specificity of novel therapies compared to conventional treatments, fostering a deeper understanding and acceptance of this targeted approach.

Collaborations with NGO

In an effort to strengthen our ties with the local community regarding cancer, we reached out to the Hong Kong Cancer Fund to establish a partnership. We engaged in a conversation through an interview and expressed our support for their mission by making donations. The Hong Kong Cancer Fund is a well-respected non-governmental organization (NGO) dedicated to offering free professional assistance and information to cancer patients and their families, extending compassionate support to those affected. They also prioritize and seize opportunities for educating the public about cancer awareness, which aligns perfectly with our objective of developing more accessible pancreatic cancer treatments and increasing public knowledge about this particular form of cancer.

The founder and CEO of the Hong Kong Cancer Fund, Mrs Sally Lo, provided insights into the various forms of assistance they offer to patients. These include family support, rehabilitation programs, and wellness seminars. She also highlighted common challenges that are often overlooked by the general public, such as the mental health needs of individuals navigating cancer recovery. Additionally, Mrs Lo pointed out the lack of awareness regarding the significance of routine health screenings.

Based on insights gained from the interview, we have identified numerous potential ways our project could contribute to ongoing efforts in the fight against pancreatic cancer.

Awareness and Education: The organization emphasized the necessity for heightened public awareness and education on cancer. Our initiative can enhance this effort by creating and disseminating educational resources about pancreatic cancer. These activities may include conducting seminars, setting up booths, coordinating community chats, and using social media platforms like Instagram and podcasts to distribute precise and current information regarding the condition.

Fundraising: The Hong Kong Cancer Fund mainly depends on public donations and fundraising initiatives to sustain its vital services. As a component of our initiative, we are initiating a campaign to sell jackets. The funds generated from this campaign will be contributed to the Hong Kong Cancer Fund, therefore offering monetary assistance for their endeavors.

Research Dissemination: The organization has shown interest in acquiring knowledge regarding the results of cancer-related research initiatives. To help this, our project can disseminate our study findings to the organization through presentations, papers, or instructive videos.

Support for Patients and Families: The organization outlined its comprehensive care for patients and families, encompassing psychiatric programs, wellness programs, art therapy, and peer support groups. Our project may support these projects by dedicating our time as volunteers, promoting these services, and facilitating connections between persons and these resources.

By forming these strategic alliances, we entered communities of individuals most needing this knowledge. The NGO, leveraging its extensive community connections and profound comprehension, assisted us in customizing our instructional resources to be culturally appropriate and pertinent, guaranteeing that our message was received and comprehended.

Conclusion

Collaborating with non-governmental organizations (NGOs) and implementing a multi-platform approach have proven successful in spreading our message to a wider audience. Our objective is to equip people with the essential information to understand pancreatic cancer, encourage early detection, and facilitate informed discussions about treatment options. In our fight against pancreatic cancer, education serves as a powerful tool, and we remain committed to continuing these initiatives to improve awareness and outcomes.

Stakeholder interviews

The interviews with Professor Chok Siu Ho Kenneth and Professor Chan Lam Stephen provided valuable clinical and research insights that significantly influenced our project. Their expertise in pancreatic cancer treatment and innovative therapeutic approaches offered a deeper understanding of the challenges faced in current medical practice. The following content presents these esteemed professionals' key reflections and knowledge. To further assess the viability of our project, we conducted another interview with Dr. Tang Kwan Ho, who offered a commercial perspective on pancreatic cancer treatment strategies. The following content outlines the knowledge we obtained from these experts.

Meet the Experts: Key Insights from Professor Chok Siu Ho Kenneth

To deepen our understanding of current treatment options for pancreatic cancer, we interviewed Professor Chok Siu Ho Kenneth, a leading expert in hepatobiliary and pancreatic cancer. This interview provided us with crucial insights into the limitations of current treatment options and the importance of developing innovative therapeutic approaches, which are highly relevant to our project.

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Pancreatic Cancer: Causes and Diagnostic Challenges

Professor Chok highlighted the cause of pancreatic cancer is often idiopathic, with no clear reason for its development. He noted that many factors may increase the risk of developing pancreatic cancer, such as diabetes, obesity and smoking. However, the lack of specific risk factors for pancreatic cancer complicates efforts to prevent the disease. Due to its deep-seated location behind the stomach, pancreatic cancer is difficult to detect early, often resulting in pancreatic cancer usually not being diagnosed until it is at a late stage. Professor Chok mentioned that targeted therapies can help address the treatment challenges posed by late detection.

Challenges in Treating Pancreatic Cancer

In the interview, Professor Chok highlighted the difficulties of treating pancreatic cancer, particularly its late-stage detection. Surgical resection offers a slight chance of cure, but for most patients present at advanced stages, surgery is no longer feasible. Chemotherapy is the standard treatment, but resistance often develops after initial cycles. Our project seeks to combat gemcitabine resistance by using dual-targeting therapies. Professor Chok’s insights reinforce the importance of developing therapies to overcome resistance and target cancer cells more precisely and efficiently.

The Role of Chemotherapy and Its Limitations

While discussing chemotherapy, Professor Chok mentioned chemotherapy remains the standard treatment for unresectable pancreatic cancer, but it comes with significant limitations. The treatment is highly toxic, expensive, and requires patient for frequent administration over several months, with regular imaging to monitor progress, which adds to the cost. A major challenge is drug resistance, particularly due to the fibrous tissue surrounding the tumour, which hinders chemotherapy penetration. As a result, patients often experience failure with first- and second-line treatments, with limited options for third-line therapy. There is also debate regarding the best first-line treatment, with Gemcitabine offering easier administration but marginally inferior survival benefits compared to Oxaliplatin-based regimens requiring hospitalisation. Moreover, Professor Chok mentioned targeting multiple pathways by using dual-targeting nanobodies could offer a solution by bypassing or disrupting the tumour’s protective barriers.

Emerging Therapies: Target Therapies and Immunotherapy

Professor Chok touched upon emerging therapies, including targeted therapy and immunology. However, he expressed concern about their current efficacy in treating pancreatic cancer due to the lack of significant side effects. Professor Chok shared the experience of a patient who experienced temporary remission through immunotherapy but suffered severe side effects, such as diabetes and nephritis. This underscores the need for more refined, personalised treatments. Target therapies aim to provide precision treatment with reduced toxicity.

Future of Protein Therapeutics in Pancreatic Cancer

Professor Chok expressed optimism about developing protein-based therapeutics, particularly precision-targeted therapies with fewer side effects than conventional chemotherapy. Therapies that target specific pathways in cancer cells have considerable potential. Dual-targeting nanobody could be part of the future of precision medicine for pancreatic cancer, offering a more targeted and effective treatment approach.

Connection Between the Interview and Our Project

The interview with Professor Chok Siu Ho Kenneth provided important insights that informed and supported our project to develop dual-targeted therapeutic proteins for treating pancreatic cancer. Professor Chok emphasised the complexity of diagnosing pancreatic cancer, which is often attributed to its deep location and the idiosyncratic nature of its aetiology. This complicates therapeutic regimens and patient counselling. He emphasised the need for innovation to create new therapies with minimal side effects and greater efficacy due to the limitations of current treatments, such as the high toxicity and resistance to chemotherapy.

Meet the Experts: Key Insights from Professor Chan Lam Stephen

The interview features Professor Chan, Assistant Dean of the Health System of the Faculty of Medicine at CUHK. Professor Chan specialises in medical oncology, particularly hepatobiliary and pancreatic cancers. Professor Chan provided many insights regarding our project, which focuses on developing a dual-targeting protein for pancreatic cancer treatment.

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Current status of pancreatic cancer

Professor Chan pointed out that the incidence of pancreatic cancer is increasing, and it is now the ninth most common cancer in Hong Kong, with about 1,000 new cases each year. He emphasised that pancreatic cancer was usually diagnosed at an advanced stage, with about 90% of patients having advanced stages and a life expectancy of less than one year. Moreover, pancreatic cancer was highly lethal and was the fourth most fatal cancer in Hong Kong, hence the urgent need for early diagnosis and more effective treatment.

Challenges in the Diagnosis and Treatment of Pancreatic Cancer

Pancreatic cancer poses a huge challenge in diagnosing and treating pancreatic cancer because of the late onset of symptoms. Patients with pancreatic cancer usually do not have any symptoms until the disease has progressed to an advanced stage. Standard diagnostic procedures include CT scans, blood tests and endoscopic biopsies. Treatment options for pancreatic cancer are limited, with only about 10% of early-stage patients receiving surgical treatment. In contrast, the majority of patients require chemotherapy to control advanced disease and improve their quality of life.

Early Detection Technologies and Prevention Measures

Professor Chan mentioned that effective early-detection methods for pancreatic cancer are still developing. Although patients with a family history of the disease may benefit from regular CT scans, current tumour markers are not specific enough for screening. He emphasised the importance of recognising subtle symptoms and seeking prompt medical attention to avoid delays in diagnosis and treatment. In addition, preventive measures for pancreatic cancer include addressing risk factors such as gallstones, chronic alcohol consumption and obesity. Professor Chan encouraged those with a family history of pancreatic cancer to be vigilant about their health and seek prompt medical attention if they experience any relevant symptoms.

The role of scientific research

In the interview, Professor Chan emphasised the importance of scientific research in understanding pancreatic tumours when this fibrous tissue prevents effective treatment. He discussed the potential of novel drug therapies and the need for better targeted therapies to improve patient outcomes. In addition, he highlighted the potential integration of technologies such as artificial intelligence and machine learning in drug discovery and treatment optimisation, arguing that these technologies can streamline the research process and improve the prediction of treatment outcomes based on patient data. Looking ahead, Professor Chan emphasised the need for early diagnosis and the development of more effective drug therapies, encouraging young researchers to keep an open mind, work hard, and continue seeking knowledge to make meaningful contributions to the field.

The Connection Between the Interview and Our Project

The interview with Professor Chan provides valuable insights related to our project on developing a dual-targeting protein for treating pancreatic cancer. Professor Chan highlights the increasing prevalence and lethality of pancreatic cancer in Hong Kong, emphasising the urgent need for more effective and innovative treatments. Professor Chan’s discussions on the challenges of late-stage diagnosis and the limitations of current treatment options underscore the significance of innovative approaches, such as dual-targeting therapies, which aim to improve patient outcomes.

Key Insights from Dr. Tang Kwan Ho

To deepen our understanding of commercial perspective on pancreatic cancer treatment strategies, we interviewed Dr. Tang Kwan Ho, an Associate Director at AstraZeneca. This interview yielded valuable commercial information regarding the ongoing clinical trials for pancreatic cancer treatment approaches, their current constraints, and potential future developments.

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AstraZeneca is a research-based biopharmaceutical company developing multiple pipelines of drugs for the treatment of cancer, including pancreatic cancer. Notable achievements include the development of osimertinib for the treatment of non-small cell lung carcinoma (NSCLC).

AstraZeneca was established in 1999 through the merger of the Swedish pharmaceutical company Astra AB and the British company Zeneca Group PLC. Astra AB originated in the early 20th century, while Zeneca was founded in 1993 as a demerger from the chemical conglomerate ICI. The merger aimed to create a more robust global entity in the pharmaceutical industry, focusing on innovative medicines in areas such as oncology, cardiovascular, and respiratory diseases. Subsequently, AstraZeneca has expanded its portfolio through strategic acquisitions and collaborations, establishing itself as a leader in drug development and research. The company garnered significant attention during the COVID-19 pandemic for its vaccine development efforts in partnership with the University of Oxford.

AstraZeneca has developed several drug pipelines for the treatment of end-stage pancreatic cancer patients. In 2019, lynparza, a PARP inhibitor, received FDA approval for the treatment of pancreatic cancer. Subsequently, the research team further developed Durvalumab (MEDI-4736, Imfinzi, Fidursi, Imfinzio), a programmed death-ligand 1 (PD-L1) blocking antibody, for the treatment of pancreatic cancer. Durvalumab is currently under clinical development by AstraZeneca and is in Phase II clinical trials for the treatment of pancreatic cancer. As a leading biopharmaceutical company globally, AstraZeneca aims to develop antibody-based or antibody-drug conjugate (ADC) approaches for the treatment of pancreatic cancer.

Connection Between the Interview and Our Project

In this interview, Dr. Tang Kwan Ho, an Associate Director at AstraZeneca, elucidated the current novel treatment strategy for pancreatic cancer. Initially, he emphasized the significance of targeting KRAS in pancreatic cancer, given that 90% of this protein is mutated. Furthermore, he expounded that targeting a single protein may not be efficacious, whereas simultaneously targeting two proteins may yield a more effective treatment for pancreatic cancer through bispecific antibody or dual-targeting nanobody approaches. Moreover, he posited that loading the antibody with a drug to form an antibody-drug conjugate (ADC) could enhance its therapeutic efficacy. Based on his expertise, we confirm the feasibility of our dual-targeting antibody design as potential treatment strategy for pancreatic cancer treatment. Following the initial success of our dual-targeting antibody, we will explore the possibility of developing it into an ADC to further improve its efficacy in the future.

Overview of the Project

According to the statistics from the World Health Organization in 2022, pancreatic cancer ranks as the 14th most common cancer and the 10th leading cause of mortality among all cancers. Pancreatic cancer is one of the deadliest cancers, ranking as the 4th leading cause of cancer deaths in the United States and the 3rd in Hong Kong. During our research on pancreatic cancer, traditional treatments like surgery, chemotherapy, and radiotherapy often fall short in almost all stages of pancreatic cancer. Current treatments face challenges such as lack of targeted therapies, and the low efficiency of chemotherapy and radiotherapy due to drug resistance, particularly gemcitabine, which leads to high relapse rates and poor survival outcomes. Our team proposes to address the molecular changes in pancreatic cancer cells that drive proliferation, metastasis, and drug resistance.

Our team named PanoPOLY is tackling the above treatment challenge by focusing on developing a novel strategy to suppress tumor growth and overcome acquired resistance to gemcitabine. Through our bioinformatic analysis, we revealed that two crucial cell surface receptors, EGFR and HER2, was significantly upregulated in pancreatic cancer cells while their levels were further increased upon development of acquired resistance to gemcitabine. Based on this observation, we hypothesize that targeting these two receptors simultaneously leads to growth inhibition and sensitization to gemcitabine treatment.

To tackle the current treatment challenges, we aim to develop a dual-targeting nanobody using a synthetic biology approach that express the peptides of EGFR and HER2 that specifically block both EGFR and HER2 and their signaling pathways. This innovative nanobody is novel, original and has not been reported before. We envisage that simultaneously blocking these receptors will result in reduced growth and increased sensitivity to gemcitabine. Our nanobody offers several advantages, including flexible design, small size, cost-effective production, and enhanced cellular penetration. This targeted strategy has the potential to overcome the limitations of current therapies and provide a more effective treatment option for this challenging disease.

Our project represents a significant advancement in the fight against pancreatic cancer, aiming to reduce the high mortality rates associated with pancreatic cancer. The success of our project could offer new hope for patients who have exhausted conventional treatment options. We aim to share our findings with the scientific community and healthcare professionals to help bridge the gap between research and practical cancer treatment solutions.

The figure below shows the stakeholders we consulted:

Stage 1: Confirming the Idea

Problem Identification

Our project emerged from the critical issue of pancreatic cancer, which has one of the lowest survival rates among all cancers. In particular, the lack of effective treatment options and gemcitabine resistance has significantly reduced the efficacy of existing therapies, resulting in high recurrence rates and poor patient survival. To address this challenge, guidance was sought from our academic advisors, Prof. Lee kin Wah Lee Terence and Dr. Wong Tsun Ting Clarence, who hold the positions of Professor and Assistant Professor, respectively, at The Hong Kong Polytechnic University, with expertise in cancer research and peptide-based cancer therapy.

During our discussion, we learnt from our supervisors that the major hurdles for current treatment of pancreatic cancer include lack of effective targeted therapy and acquired resistance to gemcitabine, which is the chemotherapy applied to unresectable pancreatic cancer patients. In addition, we learnt that it is not effective to target single target. They encouraged us to develop a dual-targeting nanobody using a synthetic biology approach that simultaneously targets two cell surface receptor proteins that are overexpressed in pancreatic cancer and further increased upon development of acquired resistance to gemcitabine. To date, there are a number of such targets reported in pancreatic cancer samples in literatures and also in publicly available datasets. The subsequent inquiry pertains to the selection of appropriate targets for the production of dual-targeting nanobody.

Initiation of the dry lab analysis

In search of receptor protein targets for production of dual-targeting nanobody, we sought for advice from Dr. Mu Quanhua, an Assistant Professor at The Hong Kong Polytechnic University, with expertise in genomic bioinformatics. With guidance from Dr. Mu, we initiated our first dry lab analysis to select two protein receptor candidates crucial for the development of pancreatic cancer with clinical significance through accessing to the publicly available datasets including GTEX and The Cancer Genome Atlas on pancreatic cancer, we demonstrated that expression of EGFR and HER2 was significantly upregulated in pancreatic cancer samples, while their overexpression was associated with poor patient survival. These findings indicate that these two receptor proteins are not only overexpressed in pancreatic cancer samples but also have clinical significance.

To further correlate these two genes in chemoresistance in pancreatic cancer patients, we accessed the database in pancreatic cancer cell lines and found that these two genes are also highly upregulated in gemcitabine-resistant pancreatic cancer cells including BxPC-3 and CFPAC-1. To further evaluate whether this observation is clinically relevant, we extended our analysis to patient data from the TCGA pancreatic adenocarcinoma cohort, in which 107 patients were treated with gemcitabine. Similarly, significantly higher levels of both EGFR and HER2 were observed in non-responders when compared to responders who achieved complete or partial remission.

Taking all these results together, we hypothesize that targeting EGFR and HER2 by a dual targeting nanobody not only suppresses pancreatic cancer growth but also sensitizes cells to gemcitabine treatment.

Stage 2: Shaping Idea

Stakeholders Interviews

To further substantiate and implement our project design, we conducted interviews with two prominent clinical experts and one expert in biopharma, all of whom possess extensive experience in pancreatic cancer treatment. These interviews provided valuable clinical and research insights that significantly influenced our project.

In our interview with Professor Chok Siu Ho Kenneth, a leading expert in hepatobiliary and pancreatic cancers, he emphasized the challenges of treating pancreatic cancer, particularly due to its late-stage detection and resistance to chemotherapy. Furthermore, he discussed the limitations of current treatments, highlighting the toxicity and limited efficacy of chemotherapy, particularly gemcitabine. After introducing our project, he expressed optimism about our dual-targeting nanobody approach, which could help overcome resistance by bypassing the protective barriers of cancer cells. His insights validated the importance of more precise therapies targeting resistant cancer cells.

Professor Chan Lam Stephen, an expert in oncology, focused on the increasing prevalence of pancreatic cancer and its high mortality rate. Professor Chan emphasized the need for early detection and more effective treatments, as most patients are diagnosed at an advanced stage with few curative options. He also discussed the potential role of scientific research and novel technologies, such as artificial intelligence, in improving treatment outcomes. His views reinforced the significance of our approach to creating a dual-targeting therapeutic protein to address current treatments' limitations.

Dr. Tang Kwan Ho, an Associate Director at AstraZeneca, who is an expert in the drug pipeline for treatment of pancreatic cancer. Dr. Tang indicated that focusing on a single protein might not be effective, but simultaneously targeting two proteins could yield better results in treating pancreatic cancer through the use of bispecific antibodies or dual-targeting nanobodies. Drawing on his expertise, we validate the viability of our dual-targeting nanobody design as a potential treatment for pancreatic cancer.

The Hong Kong Cancer Fund is a non-governmental organization (NGO) that aims to improve the quality of cancer support services in Hong Kong. It is the largest cancer support organization in Hong Kong, providing free information and professional services to all people living with or affected by cancer.

In the interview, Mrs Sally Lo mentioned that many common hardships faced by cancer patients are often neglected by the public, such as the psychological needs of cancer patients in the process of cancer recovery and the insufficient realization of the importance of regular medical check-ups. This interview taught us that our project can support current pancreatic cancer initiatives in many ways.

Mrs Sally Lo emphasizes the need to increase public awareness and education about cancer. To raise public awareness of pancreatic cancer, we have set up booths on campus and utilized social media platforms such as Instagram to disseminate information.

The Hong Kong Cancer Fund mainly depends on public donations and fundraising initiatives to maintain its vital services. As part of our initiative, our team collaborated with the Hong Kong Cancer Fund to design a jacket and organize a charity jacket sale on campus. All the profits from the jacket sale will be donated to the Hong Kong Cancer Fund to provide financial assistance for their work. The purpose of the jacket sale was not only to raise funds for the Hong Kong Cancer Fund but also to raise awareness and alertness about cancer care in the local community.

Reflection

The insights and suggestions of the three experts and the Hong Kong Cancer Fund greatly influenced and refined our project. Their contributions allowed us to identify gaps in the initial plan and address them through practical and innovative solutions. This collaboration also encouraged us to go beyond our initial scope and integrate more diverse perspectives and strategies, ensuring that our project achieves its goals and has a broader and more lasting impact. As a result, we were able to refine our approach to make the project more sustainable and responsive to real-world needs.

Stage 3: Designing the Solution

Through expert consultation and statistical analysis, we have determined the main challenges in pancreatic cancer treatment. These include a lack of targeted therapies and the emergence of resistance to gemcitabine. After the first dry lab experiment, we, PanoPOLY decided to develop a BioBrick using a synthetic biology approach to produce a dual-targeting nanobody targeting both EGFR and HER2, which showed great clinical impact to pancreatic cancer patients. We hope that our nanobody suppress the tumor growth and sensitize to gemcitabine treatment.

Our dual-targeting nanobody incorporates two distinct targeting elements: one segment targets the EGFR receptor, while the other focuses on HER2, with a connecting linker between them.

Stage 4: Conduct Experiments

After identifying EGFR and HER2, we have further consulted with Dr. Wong Tsun Ting Clarence, Dr. Wong Waiting and Dr. Lee Chun Kit Alan for the BioBrick design for our dual-targeting nanobody called Panobody. Dr. Wong Tsun Ting Clarence first guided us to design the peptide sequences that targets EGFR and HER2. To allow the flexibility of dual targeting to EGFR and HER2, a linker sequence was advised to introduce in between. Although we have this initial BioBrick design, it remains unclear whether these two peptides are specific to these two receptors.

To address this issue, we further consulted Prof. Lee Kin Wah Terence regarding the specificity of our peptides to these two receptors. He instructed us to follow the guidance of his postdoctoral fellow, Dr. Shakeel, for our second dry lab experiment. This experiment focused on protein bioinformatics and aimed to verify the specificity of corresponding targets in our treatment approach. To start with, we began by predicting the 3D structures of Panobody and a control peptide, Scramble 6, using the AlphaFold3 web server. These structures were validated using SAVES v6.1, ensuring high structural accuracy.

We subsequently retrieved the crystal structures of HER2 and EGFR from the Protein Data Bank and prepared them to use the Protein Preparation Wizard (PPW) of Schrödinger Suite 2023-1 for molecular docking. Using the BioLuminate module, we then performed molecular docking between the top performing models and prepared receptors and analyzed the resulting complexes for their molecular interactions. Upon analysis, we demonstrated the specificity of Panobody in targeting both EGFR and HER2.

Next, we consulted with Dr. Wong Waiting and Dr. Lee Chun Kit Alan regarding the BioBrick design. They both advised using a synthetic biology approach to synthesize Panobody with a BioBrick design, utilizing the pET-24d expression vector. The Panobody was tagged with histidine for purification purposes. The recombinant plasmid was transformed into BL21(DE3) competent cells as hosts for protein expression. Protein expression was induced using IPTG (isopropyl β-D-1-thiogalactopyranoside). Successful expression was verified by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Subsequently, we purified the protein by Nickel-Affinity Chromatography and confirmed the mass using mass spectrometry.

Our dry lab experiment results led us to propose that gemcitabine-resistant pancreatic cancer cells exhibit increased levels of EGFR and HER2 proteins. To validate this hypothesis, we sought the expertise of Dr. Yan Sau Woon Clare to perform our initial wet lab experiment, aiming to substantiate these findings using a cell-based model. For this purpose, we treated PANC-1, a pancreatic cancer cell line with increasing doses of gemcitabine continuously for more than 2 months, and verified the successful establishment of gemcitabine-resistant pancreatic cancer cells using the MTT assay. Following the establishment of these resistant cells, Professor Lee Kin Wah Terence instructed us to assess the cell surface expression of both these cells and their mock control counterparts using flow cytometry analysis. Subsequently, under the guidance of Dr. Carmen Leung, a postdoctoral researcher in Professor Lee's lab, we evaluated the effectiveness of Panobody, both alone and in combination with gemcitabine, on gemcitabine-resistant pancreatic cancer cells. This evaluation was conducted using an MTT assay, followed by Bliss analysis.

Stage 5: Outside the Lab

We conducted various initiatives to promote awareness about pancreatic cancer and synthetic biology. Initially, we distributed a questionnaire to assess public knowledge of pancreatic cancer and shared Instagram reels along with weekly quizzes. As our experimental work progressed, we expanded our outreach efforts by hosting additional workshops and setting up informational booths to showcase our project and educate people about synthetic biology.

Moreover, we held a workshop to provide secondary school students with an introduction to fundamental concepts of synthetic biology and biotechnology. Specifically, we separate our workshop into two sections: the educational presentation part and the interactive activity’s part. As for the educational presentation, we summarized our projects related to pancreatic cancer and some concepts about synthetic biology. For interactive activities, we played games such as bingo games, board games, and Q and A discussions to let secondary school students have a better understanding of pancreatic cancer and synthetic biology. Additionally, PanoPOLY joined the APAC Mock Jamboree as the cohost to promote our project and communicate with other teams.

Furthermore, we participated the 57th Joint School Science Exhibition, which is faced to the public. Instead of only having an introduction to our projects in the booth, we also printed leaflets with pictures to help the public better understand pancreatic cancer. Moreover, based on our project, we have designed a science mini board game, in which players should use specific drugs to treat specific cancers. In addition, we held an education booth at our university campus and gave education presentations for university students during the orientation period.

Finally, we interviewed Prof. Chok and Prof. Chan to absorb more valuable knowledge in clinical and research insights that significantly influenced our project on developing a dual-targeting protein for treating pancreatic cancer. After the interviews, we did a collaboration with the Hong Kong Cancer Fund and conducted an interview with the founder of NGO. For the fundraising, we have designed a jacket to raise funds and will donate all the profits we made to the Hong Kong Cancer Fund.

Our team also took part in the 57th Joint School Science Exhibition, an event open to the general public. We not only provided information about our projects at our booth but also distributed illustrated leaflets to enhance public understanding of pancreatic cancer. Additionally, we created a science-themed mini board game based on our project, challenging players to use specific medications to treat various cancers. We also set up an educational booth at our university campus and delivered informative presentations to university students during their orientation period. As mentioned in the earlier section, we also conducted interviews with Prof. Chok, Prof. Chan and Dr. Tang to gain valuable clinical, commercial and research insights for our project on developing a dual-targeting protein for pancreatic cancer treatment. Following these discussions, we collaborated with the Hong Kong Cancer Fund for charity merchandise.

Here is the roadmap for our Human Practice activities:

Conclusion

Throughout PanoPOLY's project journey, our achievements were primarily attributed to expert guidance and unwavering dedication. Key stakeholders played a crucial role in steering and honing our progress, while our commitment drove continuous enhancements. The collective effort of our team resulted in the successful development of a groundbreaking Panobody, which shows promise in inhibiting pancreatic cancer growth and complementing gemcitabine treatment. Our ultimate aim is to enhance therapeutic efficacy and boost patient survival rates. This accomplishment stems from collaborative expertise and our relentless pursuit of advancing cancer treatment innovation. We are confident that PanoPOLY's innovative approach has the potential to improve the curability and survival outcomes for individuals battling pancreatic cancer.