During the brainstorming process, we wanted to see whether the process of using synthetic biology to produce ursolic acid was really viable and the most efficient option. Dr. Mikail Abbasov is a chemical biologist developing new chemistries to understand how mutations and post-translational modification alter protein function and lead to disease. We reached out to him because of his expertise in small molecule drugs. Dr. Abbasov’s interview was immensely helpful because he advised us that ursolic acid synthesis is a complex enough process that biological manufacturing is recommended. He also revealed another benefit of the Oncurex enzymatic process: the fact that it does not include any toxic intermediates.

Take Aways: Our project has merit and is promising. The only method that makes sense for extracting the acid is synthetic biology due to its complex process and has no toxic intermediates.

To investigate the merits of the Oncurex project, we interviewed Josh Wong. Josh Wong is a chemical biology PhD student studying natural product chemical synthesis. From talking with him, he revealed that the Oncurex biological process is more industrially viable than synthesis. This is an exciting fact to notice, as it reiterates that we do not need to be worried about there being another cheaper alternative. He said that you would not be able to synthesize ursolic acid without an enzyme and that this is a good example of a molecule that a model organism would be helpful to synthesize for clinical purposes. This meeting left us understanding that the Oncurex route of producing ursolic acid using synthetic biology is an extremely viable route.

Take Aways:Producing ursolic acid using synthetic biology is an extremely viable route and we should not be worried about there being different, cheaper alternatives. This also reinforced that we should use yeast as our model organism as it already creates the precursors and enzymes to ursolic acid.

During our brainstorming project, issues arose about the separation process for ursolic acid. Before continuing to pursue this idea, we wanted to make sure that this process was viable, and so we interviewed Dr. Kevin Sigenthaler, a molecular biologist with expertise in genetics. He informed us that the ursolic acid would likely be extracted through a lipid extraction and that the extraction process should be more simple because the ursolic acid is not found in the cytosol. He also talked about the benefits of using yeast and the ability of yeast to work well for scale-up. He opened our eyes to the fact that we are going to have to have a mechanical lysis to break down the yeast, so we realized we needed to do more research on that topic. Ultimately, Dr. Siegenthaler’s insight told us that our idea was valid, but that we needed to dig deeper into how we planned to remove the ursolic acid from the yeast cell walls.

Take Aways:We need to do further investigation into how to remove the ursolic acid from the yeast cell walls, but the likely option is by removing it through lipid extraction. This also emphasized that yeast is a good option for this process as it will be easy to scale it up.

After deciding to pursue Oncurex as our final project, we reached out to Agnes Slawska to talk about the logistics of having the product enter the market and to discuss potential ethical concerns with the project. Ms. Slawska works as the US Marketing Lead for Agios Pharmaceuticals. She urged us to make sure to look at the specific instances where the ursolic acid had worked to really push through the idea that it has very promising effects for different cancers while marketing the product and reaching out to companies. She also touched on some ethical implications with the project such as ensuring that the product is available for all that need it and determining how to make that possible.

Take Aways:We need to consider the ethical implications of our project and to focus on the specific instances where ursolic acid has been successful so far. We should also make a change in our policy/ethics framework to include the ethical changes she advised.

Dr. DeLisa’s research focuses on understanding and controlling molecular mechanisms in protein biogenesis. Specifically, he studies the folding, assembly, membrane translocation, and post-translational modifications of proteins. He has created various commercial products to manufacture human drugs. His lab works to bridge biology with chemistry by manipulating cells. We were referred to Dr. DeLisa by multiple engineering professors as a possible contact for bioreactors. During our conversation we spoke about both Wet lab and Product development to gain a deeper understanding of both genetics and the creation of the bioreactor. For the purification of our product, he recommended we first find where UA accumulates. If it is outside we can find what molecules it is attracted to and use this to draw UA out and purify it. In relation to prolonging the lifespan of yeast he recommended we either utilize continuous cultivation of the cells or grow the cells in a fed batch, as reengineering the yeast ti do this may prove to be difficult. On the PD side, he recommended setting up a simple tank CSTR as it will most likely be cheaper and using a magnetic stir bar to keep the tank continuous. He said the biggest issue we would face was contamination especially since we are making a continuous system. Additionally, to add in a biosensor he recommended doing it outside of the reactor, since doing it inside would most likely be too complicated.

Take Aways:Dr. DeLisa recommended either continuous cultivation or fed batch growth, along with a simple tank CSTR. We said a main concern would be contamination, and that a biosensor would need to remain outside the reactor. Overall, we will use DeLisa’s advice when setting up our bioreactor to ensure it is something our team can most likely accomplish while still propelling our project further.

Dr. Jiang is a biomedical engineer at Cornell University who focuses on creating drug delivery systems for cancer vaccines and neurological diseases. Currently, his lab is investigating extracellular vesicles released by cells that contain stem cell materials. His lab injects these EVs with mRNA in hope of studying regenerative medicine. During our meeting Dr. Jiang helped us investigate the use of Lipid Nanoparticles in our project. Together we came to the conclusion that using a LNP delivery system would not make sense in the context of our project. Instead, since Ursolic Acid is a small molecule, we should use liposomes. As we talked further, he gave us several recommendations on how to make a more innovative project. Specifically he referenced his research and use of extracellular vesicles. He encouraged us to explore yeast and how they produce EVs to see if we could use this as a delivery mechanism for our drug. He informed us that all products of our yeast should be encapsulated into the EV and we should measure how much UA is present in each one. Although purifying the product may be difficult, he recommended we use yeast that are not harmful to humans or possibly shift the result of our project to be agricultural based if the yeast EVs would threaten human health. He told us that EVs are naturally programmed to go to specific locations and these locations may be cancerous, so it may be worth it to track where yeast EVs go naturally before trying to edit or complicate the system.

Take Aways:Dr. Jiang recommended use of yeast extracellular vesicles and the use of liposomes as a potential pathway. He encouraged us to consider tracking where the extracellular vesicles go naturally before editing the system, as well as the benefits of different applicable fields such as medicine or agriculture.

Dr. Specht is a Cornell University postdoctoral student who is focused on researching the transformation process of Vibrio natriegens. There has been lots of research done in Vibrio cholera, but Dr. Specht focuses on natriegens and trying to extract methods from cholera to apply them to these specific bacteria cells. He has created various novel ideas surrounding these bacteria and ways to manipulate them in a more cost effective and efficient ways. We asked him if he thought we should use Vibrio natriegens as an intermediate in our pathway to make everything more efficient and novel. He recommended that we just introduce our plasmid to the yeast cell directly as yeast already have an established pathway, so our plasmid will be small. Additionally, Dr. Specht has spent a lot of time using the Gibson assembly technique. We expressed to him the difficulties we have faced with the technique and he said one of two things could be going wrong. One possibility could be that our PCR was too cheap and did not have the correct materials to allow the path to occur. Additionally, we could not be expressing or making what we originally thought we were. The second possible error could be in the actual design as it may be toxic. A lot of time bio bricks can be very strong and this strength can kill proteins on accident. He recommended starting off slow with a weak ribosomal binding site and promoter and then increasing strength from there.

Take Aways:We will definitely take Dr. Specht's advice into account, especially since we have had difficulties with our Gibson assemblies in the past. He gave us an abundance of information to make sure our assemblies are going smoothly. He also said he would be happy to have us in his lab to show us some techniques to make our methods go smoother and this is something we are planning on looking into further.

Professor Fromme is a principal investigator for both the Weill Institute for Cell and Molecular Biology and the Department of Molecular Biology and Genetics at Cornell university. His research focuses on the Golgi Complex and its function of sorting within the cell. His lab was able to discover how GTPase is activated in order to regulate these mechanisms within the Golgi. Our primary focus to reach out to him was his use of yeast. Our team is not familiar with the organism and we wanted to go over specific protocol regarding them. Professor Fromme was able to give us a lot of good advice and material on working with yeast. He assured us that working with yeast would most likely not be an issue as it should not pose a safety threat. Unless one of our members is severely immunocompromised there should be no big issue working with yeast. To clone yeast his lab deploys methods of recombination when introducing plasmids. He also said that having more plasmids in the yeast will increase the overall yield of the acid while also making sure to select a good promoter. Additionally, he said we may not need to worry about yeast aging and that our culture can most likely be stored in the freezer for long periods of time without the yeast dying. He also shared multiple different resources and protocol books with us.

Take Aways:Our team is looking forward to using his input and advice to make sure our project is safe and that we can extract Ursolic Acid form the yeast. Professor Fromme also provided us with some of his yeast so we can grow them and insert our plasmids in them. We also took away key components of his handbook which allowed us to understand how to work with yeast, as this was the first time our team used yeast as a model organism.

Our second conversation with professor Fromme went over our plasmid design. He told us that there are various ways to design the plasmid that is more effective. One way would be to use primers and a plasmid with the alpha amyrin template cDNA already within it. Another way to make it more effective would be to cut the Ura3 site, insert alpha amyrin synthase, and then perform the POR2 knockout in a separate region of the plasmid. An additional way to better the plasmid would be when the Ura3 site has a point mutation. When this occurs you can perform a single crossover where you duplicate the Ura3 site and then the gene of interest is inserted in between the duplicated Ura3s. It is also important to note that primers are essential for validating these constructs. He also discussed how to get yeast to pick up the new plasmid. One of his suggestions was to have yeast take up various parts of the plasmid and integrate it into their own DNA with homology regions. He explained that yeast are very recombinant organisms.

Take Aways:We will apply his advice moving forward by trying multiple of these methods in parallel to see what works for us. He gave us the freedom to explore multiple methods not only designing our plasmid but also how to get the yeast to actually use our plasmid in the most efficient manner. Multiple different methods can be used at once and we must find a collection of methods that have the correct harmony to produce the most ursolic acid.

Dr. Van Campen is the CEO of EthicsMatters, a bioethics company, and has extensive experience in bioethics, organizational ethics, and research due to her diverse previous jobs across various sectors. We met with Dr. Van Campen to discuss ethical considerations that we should have about our project. She emphasized the importance of thinking of ethics early and often, and also elaborated on the four components of ethics: justice, autonomy, non-maleficence, and beneficence as well as their relationship to our project. She brought up the issue of drug delivery, and proposed that when deciding the type of delivery method (supplement or injection), it would be important to consider beneficence and justice mostly to ensure that the populations that are most in need of this treatment can receive it in the most efficient way possible. This interview was instrumental in our project process because it led us to more deeply analyze which method of drug delivery would be most optimal. Our interviews with Dr. Van Campen also encouraged us to pursue leads at insurance companies to see if ursolic acid would have potential to be approved by insurance companies and thus be increasingly accessible.

Take Aways:There are four components of ethics: justice, autonomy, non-maleficence, and beneficence, and we should consider all of them in our project. We should reach out to insurance companies to see if they would approve ursolic acid as a drug. We should consider how the ursolic acid would be delivered.

Dr. Jini Hyun is an Assistant Professor of clinical medicine at Weill Cornell and an attending physician at NYPQ. She has a background in Pharmacology and completed an Internal medicine residency at NYPQ. During this time, a large portion of her research focus surrounded the herbal extractions done in Korea and China in old literature and pharmacological tracing to make pure compounds. She believes creating a cancer care plan is complex and requires a team with the patient being at the forefront of decision making. She also thinks it is important to create a personalized plan with specific goals of care to maximize the quality of the patients' lives. Specifically, she has used genomics, molecular biology and therapeutics over the last few decades to treat many of her patients. During our conversation she spoke a lot about the extraction of specific medications from plants. She was excited to hear about our plan to extract this natural and medicinal acid in a way that was environmentally conscious. She also spoke about the process of clinical trials and what we can expect as ursolic acid goes through these trials for cancer. She was not too concerned about the amount of time it is taking for ursolic acid to move to the phase two trials but she urged us to email the PIs to see how their current research is going now. As a physician, she said if Ursolic Acid was FDA approved and part of a regular treatment plan she would be happy to use it as a medication.

Take Aways:She was excited to hear our project, and explained to us the process of cancer treatments and where Ursolic acid would fit in. She recommended us to continue to reach out to oncologists and ursolic acid specialists regarding our project and even ask the NIH and FDA about the process of clinical trials. She urged us to speak to researchers to gain a better understanding of the current phase one trials of ursolic acid as a cancer therapeutic.

Dr. Ran Yin is currently the CTO of a Chinese biotech company, BIOEVERYDAY. He was previously a PhD candidate in the Liu Lab at Cornell, conducting research focusing phytochemicals from fruits and vegetables in regulating cell proliferation and apoptosis in human breast cancer cells. He found that UA had the most promise out of the phytochemicals he studied for its anticancer and antioxidant properties. It has a lot of flexibility and can be used as dietary supplements or synergistically with other therapies. At the molecule level, UA has several targets on cancer cells that can be maximized for anti-cancer effects. He highlighted several challenges to the engineering and clinical implementation of UA: 1) it has a high antioxidant activity that may damage yeast, 2) as a large hydrophobic molecule, it is hard for humans to absorb an active concentration, and 3) very high amounts of UA can have cytotoxic effects for healthy tissue. There is a window for an active concentration against cancer while also avoiding normal tissue toxicity.

Take Aways:We will investigate the antioxidant activity of ursolic acid in engineering the yeast and look into an optimal active concentration of ursolic acid. He said that UA has the most promise specifically for cancer applications, but highlighted other applications and several challenges that prompted additional interviews, Cornell iGEM decided to integrate the different applications of ursolic acid into the Ursolic Acid Historical Handbook as a second component based on the interview.

Dr. Susan Sadoughi works at Mass General Brigham and specializes in internal medicine. She has a lot of experience helping patients with their treatment plans and has an in-depth understanding of the process of drug approval, so we spoke to her about how patients and doctors decide on treatment plans and enroll in experimental drug trials and the various applications of ursolic acid. She explained that the patients in phase I trials are only those who are already projected to die, and that they engage in these trials to help science. Phase II and phase III, however, can be enrolled in by the patient if the typical treatment plan is not working so the specialist will suggest that a trial could be helpful. She described that, when creating a treatment plan, doctors normally first look at what type of cancer it is, what stage, and then how aggressive the cancer is. Based on that, she described that there are generally not many options for cancer treatment, especially compared to other conditions, but that to decide on a final treatment doctors look at the patient's medical history and the potential side effects and how willing the patient is to tolerate the side effects. When discussing UA application to other fields such as viral medications, Dr. Sadoughi stated that based on the research a cancer treatment is more comparable that antiviral. Dr. Sadoughi expressed concern with the fact that ursolic acid has been stuck in phase I trials, and encouraged us to find out why the drug has not moved onto phase II trials. This led us to reach out to some of the authors on the scientific papers on ursolic acid to find out the reason for the hold up.

Take Aways:We learned about the process that doctors and patients take to create a treatment plan and how patients and doctors decide to enroll in clinical trials / to try experimental drugs. We decided to find out more about the lack of phase ll trials, and also pointed away from specific ursolic acid application processes such as anti-viral applications.

Dr. Joshua Zaritsky is a pediatric nephrologist who specializes in treating rare diseases. He often deals with children's cancer, and is familiar with the process of clinical trials. We consulted him to get feedback on the general cancer treatment process and the process of implementing a drug into treatment from clinical trials. He said that he normally only consults clinical trials when other, more established treatment methods have been failing, especially because he is dealing with children. When asked about the drug only being in phase I, he also expressed significant concern about this and said that it was integral that we look into the reasons that it is stuck in phase I. This prompted us to reach out to even more ursolic acid researchers. He encouraged us to explore some of the other applications for ursolic acid because he said it could have significant promise outside of cancer applications. While he stated that UA does not seem as applicable to kidney diseases, he touched on how, even if ursolic acid does not end up having efficacy for a cancer or kidney specific application, it is still a useful compound and that our method of producing it more efficiently and sustainably is helpful.

Take Aways:Dr. Zaritsky pointed out concerns that UA has not been researched enough in kidney applications, but stated in terms of cancer treatments that UA could be still a useful compound and our method of production is promising. We should continue investigating why the ursolic acid has not progressed into phase II trials, but our project idea in general is promising.

Dr. Saxena is an Assistant Professor of Medicine at Weill Cornell Medical College and Attending Physician at New York-Presbyterian Hospital. He works at Weill Cornell in the Division of Hematology and Medical Oncology with a concentration in Thoracic Oncology. He continues to study the different forms of lung cancer and treats patients with various types of lung cancer. Dr. Saxena is primarily a medical oncologist and spends most of his time focusing on treating patients with specific therapies rather than surgery or radiation. We spoke primarily about how specific treatment plans are chosen for patients and the side effects of these plans. We learned about the three major treatments for lung cancer including chemotherapy, immunotherapy, and small molecule therapy. He also spoke about the side effects for each therapy and how additional medications need to be prescribed to alleviate these side effects. He was concerned about the amount of time Ursolic acid has been stuck in the clinical trials and said it could be because of the functionality of the medication or the cost. He advised us to continue looking into the trails and Ursolic Acid as a whole. He also reminded us to find a specific subset of cancer that the acid could be applied to. He said it was easier when the treatment group is narrower as the treatment works better than when giving it to multiple people with different backgrounds and types of cancer.

Take Aways:Dr. Saxena was able to provide an abundance of information on current cancer treatments and their effects on the human body. He was also able to break down the clinical trial process in America and help us understand the importance of making our project target more specific. This helped us to look into all of the cancers the acid has been used on and choose our focus.

Dr. Rajiv Magge is a professor of Neurology at Weill Cornell and a Neurologist at NYPH. He specializes in neuro-oncology and gives care to patients with brain tumors and people who have suffered neurological damage from cancer. When speaking with Dr. Magge wanted to gain more information about the process of clinical trials and his opinion on the trials of Ursolic Acid. We were also hoping to gain more insight about current cancer treatments. He gave us plenty of information on various cancer treatments like chemotherapy, immunotherapy, target agents, radiation, and surgery. He also helped us better understand the side effects that accompany many of these treatments so we can see if these specific side effects would be lessened with the use of Ursolic Acid. Dr. Magge also brought up some concerns he had about the clinical trials occurring with Ursolic Acid. He found some contradictory points within the paper and noted that the study was being done in China– a country with much fewer regulations in comparison to the USA. He advised we take the study with caution and try to learn more about the trials being done. He also explained that many molecules do exhibit anticancer properties. Many times when molecules are placed in a petri-dish with a cell they are able to kill cancer cells; however, this rarely is able to translate to actual anti-cancer mechanisms in vivo. He emphasized the importance of making sure that Ursolic Acid is actually a reliable drug in vivo as well as in the beginning stages of trials. He also encouraged us to fine tune and specify the type of cancer we want to focus on. Cancer treatments have long moved on from blanket treatments and now need to be highly specific to the cancer being treated. As a team, we are hoping to take Dr. Magge’s advice and find more information about the clinical trials. We also want to begin to fine tune the type of cancer we will target with Oncurex.

Take Aways:We are taking away a better understanding of how the ursolic acid trails are undergone and the possibilities of errors that could have been encountered.. He also brought up concerns about the current studies which urged us to continue investigating the trials to get our answers.

Cornell iGEM met with Dr. Blessing Aderibibge, an ursolic acid researcher specializing in biological applications. Dr. Aderibibge presented to us the various applications of UA, along with specific details on its biochemical makeup. Specifically, we learned that currently UA is classified as a Class lV drug due to its poor oral bioavailability. It is difficult on its own to use as a drug due to its poor solubility, poor bioavailability, and poor membrane permeation. It is difficult to administer orally as well, as it is not properly absorbed. Administering UA through an injection however leads to rapid absorption in abundant blood supply organs. One way this issue of bioavailability has been addressed is using modified UA. Specifically, after modifying UA either with esterification, amidation, glycosylation, oxidation/reduction, or acylation, the drug performed better than UA alone. As such, UA modification is necessary for an enhanced biological effect. Several reports have shown that increasing solubility, bioavailability, and potency of UA can be done with modifications at different positions. She stated that cancer is the most promising path related to UA, but it is unknown the exact biological mechanism UA uses to differentiate between cancer cells and non-cancer cells. Dr. Aderibibge states that scale up of production of UA is not a problem and that upscaling UA production is not helpful, but the main issue for it to be used biomedically is to increase modification or using nanocarriers to incorporate UA.

Take Aways:Dr. Aderibigbe offered a lot of extensive information on the applications of UA as well as its biochemical properties. She outlined two types of Ursolic acid modifications, such as chemical modifications or the implementation of nanoparticles. Both paths offered potential, so future interviews investigated the viability of both.

We met with Dr. Sean Nicholson, a professor of policy analysis and management, and the director of the Sloan Program of Health Administration at Cornell University. Dr. Nicholson discussed the potential behind our project in terms of management and potential stakeholders/consumers. Dr. Nicholson said that There are two potential values to our project: 1. whether we will be able to produce something that is either lower cost or produces more, and 2. Whether our project touts environmental progress. He noted that many companies are becoming increasingly conscious of environmental concerns, but it is now a matter of balancing between whether companies would prioritize an environmentally friendly product over cost and efficiency. He gave us direction to reach out to Contract Development and Manufacturing Organizations that support pharmaceutical companies, as well as how we should differentiate our Ursolic Acid from others currently on the market. He also noted that it does not matter as much whether pharmaceutical companies are undergoing phase 1 trials and beyond, as they need medicine to conduct the trials and our project fills that need. Overall, he applauds this project for being bold, and encourages us to consider all options and pivots necessary.

Take Aways:Dr. Nicholson offered key advice on the value of our project to potential biopharmaceutical company consumers, and direction regarding project values and outreach.

We spoke with Dr. Bruce Ganem, an Organic Chemistry professor here at Cornell University regarding potential modifications to Ursolic acid, following a lead by Dr. Blessing Aderibigbe earlier. Dr. Ganem stated that there is little difference to chemical modification of UA that makes it distinguishable from other common chemical synthesis methods. In addition, chemical synthesis is not environmentally friendly and will significantly raise the environmental factor of our work, detracting from the original goal of sustainability. Overall, while he approves the innovation behind the work, he highly suggests a different approach than chemical modifications as it is not viable for the project.

Take Aways:Dr. Ganem pointed out that chemical modifications are standard and not particularly unique in the industry. Additionally, it would raise the E factor of the project, detracting from the original goal of a sustainable, environmentally friendly and efficient alternative to current methods.

We met with Professor Ying Yang, the associate director for the center of life sciences ventures and a former McKinsey consultant about the business behind our project. Professor Yang stated that the main concern she had was with the exact problem we were looking into, and told us to hone in upon the specific market we need to focus on and how UA is currently used. She noted that since ursolic acid is already commonly found in nature, our best bet would be to focus on the modification component as well as the knowledge that our project is applicable to other natural compounds. She gave us great direction in terms of honing in on the market need of our project as well as specific paths we should look into regarding the cost and need for scale up.

Take Aways:Professor Ying Yang pointed out some concerns with FDA approval and marketing, pointing us to different directions on how to understand and answer these questions.

Elizabeth Kim is the General Counsel for Mitsubishi Tanabe Pharma America. She is an expert in FDA regulatory and compliance procedures for new pharmaceuticals. She highlighted that evidence from Phase I, II, and III clinical trials are required in order to create a submission for FDA approval. She discussed that there are many reasons why a pharmaceutical may remain in Phase I trials, notably if there are issues with safety and efficacy of the product. She also noted that the clinical trials do NOT need to be conducted in the United States as long as they adhere to international and FDA standards for clinical trials. This is particularly helpful to know as clinical trials for ursolic acid, as well as the majority of in vitro and in vivo studies have been conducted internationally. For example, Mitsubishi Pharma has received approval for drugs that they have developed through clinical trials solely in Japan, which adhered to international standards for clinical trial practices.

Take Aways:She highlighted the need for clinical trials for FDA approval, and the various possibilities for clinical trial barriers. She pointed out that clinical trials are not tied solely to the USA, and can be applied to trials across seas.

We met with Dr. Carey, a health insurance policy specialist regarding the applicable federal regulation components of our project. She stated that it looks like our project can address a key issue with botanical drugs, as our Ursolic Acid product currently falls under that category. She stated that botanical drugs are commonly heterogeneously sourced, creating huge issues in standardization of medicinal output. Through a synthetic biology method, our project gets rid of issues with quality standardization and control. She also pointed us to a drug repurposing standpoint. In addressing an issue brought up by Dr. Ying with patenting, she noted that our work can fall under a Method of Use patent instead of a patent on the active ingredient itself, and applauded our decision to pursue a provisional patent on the process. Overall, she greatly encourages our work, and pointed us to both new and old directions based on supplier demand and project needs.

Take Aways:Following up on the Prof. Yang interview, we learned about one of the main issues of FDA approval from a policy standpoint, and elaborated upon routes of marketing and IP protection.She was extremely encouraging at the prospect of our project being a way to address the issue of botanical drugs, and highly encouraged us to continue our research. This was subsequently integrated into our project as a response to concerns on FDA approval.

We met with Dr. Oyedeji, who answered many key questions on our project. Regarding clinical trials and entering phase 2, he noted one of the biggest problems in preventing phase 2 trials is that modifications of UA takes too much resources and time, which delays phase 2 projects. In combination with this, nanoparticle applications could help with mitigating the time and energy it takes to modify UA. He also explained the barriers of UA approval for medical treatments as being largely based on solubility and toxicity concerns. While UA has very low toxicity, it does have a very fine line between toxicity and non toxicity, as well as difficulties in overall bioavailability. In addition, he explained to us the current process of extracting UA from plants, and how it is very time consuming and inefficient. He noted that our project has huge implications for UA research, as current UA on the market is very expensive and is actually a prime reason many UA researchers synthesize their own UA instead of buying it, which creates potential issues in standardization.

Take Aways:Dr. Oyedeji explained to us the main reason behind the minimal phase 2 trials present, and elaborated that our project still has multiple applications even barring phase 2. He also elaborated upon the plant extraction method for Ursolic Acid and how our project addresses multiple present concerns regarding cost and efficiency.

Hannah Bogich has experience as a pharmaceutical researcher and in overall drug and product research and development. We spoke to her to learn more about nanocarriers and the potential of beta cyclodextrin and other nanocarriers for our project. She explained that the company which she worked for, Dicerna Pharmaceuticals, differentiated themselves by not using nanocarriers and rather adding an ending cap to DsiRNA that helped it travel through the bloodstream to the target area. They chose to opt for this method rather than nanocarriers because nanocarriers have the potential to be toxic or to reach the wrong part of the body. While she explained that she did not have enough knowledge to speak on the efficacy of beta cyclodextrin for our project specifically, she was enthusiastic about the idea that we were not using nanocarriers but were trying to find an adjacent method to achieve the same end goal of improved drug delivery.

Take Aways:Mrs. Bogich recommended the use of an alternative to nanoparticles through the use of beta-cyclodextrins per prior literature into Ursolic Acid. She stated that nanoparticles can be toxic, and that our new method of beta-cyclodextrins is a promising alternative.

We met with Dr. Lewenstein, a public engagement and scientific communication professor at Cornell regarding our project and outreach events. Dr. Lewenstein was very enthusiastic about our work, and was particularly encouraging of the IRB and children’s book. He explained to us how scientific communication interacts with public engagement and science education via a venn diagram, and detailed the need for more understanding of scientific communication. Dr. Lewenstein also explained to us a project he worked on regarding cancer patient perspectives, and explained different demographics that we should continuously keep in mind during our project. He recommended that we continue our current trajectory, and pointed us to next directions on cancer patient interviews.

Take Aways:Dr. Lewenstein elaborated upon different methods of scientific communication, encouraging us to continue our work in forming an IRB to speak with patients and alternative methods of outreach such as the children’s book. He pointed us to other researchers who have worked with cancer patient communication in the past.

We met with Dr. Robert Weiss, a researcher at the Cornell Vet School who has organized patient interviews in the local community. Dr. Weiss was extremely enthusiastic about our project, and stated that patients will give a nuanced perspective and insight that other interviews will not bring. He stated that it will be extremely helpful to define the interview group as breast cancer survivors pertaining to our project. Dr. Weiss additionally gave recommendations on how to approach cancer patient interviews, focusing on the patient’s story and not over-promising based on the scope of the project. He gave local community initiatives to connect to as well as elaborated on how to establish rapport with patients beforehand.

Take Aways:Dr. Weiss pointed us in various directions, and put us in contact with multiple organizations around the Ithaca local community to conduct outreach and learn more from the perspective of cancer patients.

Dr. Raymond Glahn is a visiting Professor at Cornell University. He researches the ways to enhance certain types of nutrients into crops. Specifically, He has worked with enhancing the iron in beans and various projects relating to Mazoie. By enhancing the nutrients people are able to combat certi9an diseases by having enough vital nutrients and minerals in their body. We wanted to speak to Dr. Glahn about biofortification to see if this would have been a viable route to our project. Through talking through our approach we learned that Biofortification is very time consuming. It cannot be done on all plants and does not even necessarily guarantee an increased yield when extracting Ursolic Acid from plants. This process would take a minimum of 8 years and require a large amount of resources. Additionally, even if this type of proceeding would work on loquat plants or apples to increase the amount of ursolic acid in its fruits it may not have an equivalent expression every year. This would mean some years people would not be able to extract meaningful amounts of ursolic acid. This further proves the point that fruit extraction is an unreliable source of extraction as the level of UA in each fruit could have a large range. He also said the biggest tool to combat misinformation of GMOs or plant breeding is through education. He said everyone may need different levels of education but it's important to have a large reach.

Take Aways:Dr. Glahn gave us information about the biofortification of plants, so we could see if it would have been a viable alternative as recommended by various outreach events. Through this interview, we learned that biofortification is both time and resource consuming, and unreliable as a whole.

We spoke with Sophia Openshaw to investigate the necessary steps in calculating the environmental factor of our project. Sophia Openshaw has previous experience in calculating the impact of making changes to more sustainable technologies, as she has experience creating life cycle analyses of biodiesel and diesel fuels to compare their environmental impact. She suggested that we look for papers that have already calculated the environmental impact of similar processes to make an estimate for the environmental factor of our process. To compare it to traditional processes, she suggested that we make rough estimates based on the environmental impact of producing apples and the portion of apples that are used to produce ursolic acid.

Take Aways:Mrs. Openshaw helped explain the impact of sustainable practices upon our project, and gave us directions in calculating the environmental impact using E-factor.

Dr. Li is a professor at Cornell and a member of the USDA-ARS as a researcher in molecular biology. Dr. Li focused on the nutritional quality of food crops through the focus of gene discovery, micronutrient nutrition, biofortification of plants in the context of nutrition, and plant biotechnology. We wanted to speak with Dr. Li to gain more insight on the manipulation of plants and if this is a viable process for expressing ursolic acid. She was able to walk us through the biology of plant manipulation and highlighted the importance of finding each metabolic pathway, the key elements within them, and their precursors in order to be able to express a compound within a plant. She said that this could be done fairly quickly in wells studies model organism plants; however, apples and loquats do not fall under this category. She also informed us that for every molecule plants have an internal threshold of how much they can produce until they die or begin to fight back and regulate the production of the molecule. She also explained to us ways we can inform people about genetically modified organisms to make them feel less scared. She said it is important to prove the safety of what you create to put people's minds at ease.

Take Aways:Dr. Li helped us understand the process of biofortification better by thinking about key regulators and enzymes on the cellular level. This interview helped us understand that modification of yeast is the most efficient and least time consuming method. She also helped us understand how we can start to break down the stigma against GMOs with education.

We met with Bob and Monica, cancer advocates who have previously worked with the Cancer Resource Center in the Finger lakes. They both have past experience both with speaking with Cancer patients, and with organizing and leading focus groups. They highlighted key points of the cancer treatment experience, and pointed out many myths associated with the healing process such as being cleared after 5 years. Bob and Monica additionally shared common phrases and sayings they suggest we avoid as that diminishes to experience, such as the description of cancer being a “battle” and that “they are so brave”. The overall emphasis that they made was on the unique story that each cancer patient has to share, and to really focus on listening and understanding instead of reacting. Bob and Monica also aided in revising our interview process for cancer patients, and conducting mock interview training with our iGEM members tailored to the cancer patient and their lived experience.

Take Aways:Bob Riter and Monica Vakimer helped us reevaluate our questions for cancer patient interviews within our IRB, and led mock interviews for interview training specifically for patients. They highlighted key points in the patient experience, and led us to specific ways to learn from patients.

We met with Dr. Ben Cosgrove, the Director of Graduate Studies at Cornell’s Meinig School of Biomedical Engineering. We proposed our modelling approaches to Dr.Cosgrove and he explained that they are good justifications for the wet lab approach. In particular, he believes that the Michaelis-Menten modelling is accurate in scope and provides good information on enzymatic dynamics and pathway sensitivity. In terms of the network topology analysis, Dr. Cosgrove explained that a flux balance analysis (FBA) might be better suited to our goals. He explained that a rigorous FBA would provide more information as graph theory is too static. However, after explaining our approach with the thermodynamic weighting of edges, Dr. Cosgrove explained that this is an acceptable alternative. He suggested that we edit our documentation and theoretical development to match our approach. He also suggested that we consider even more variants of alpha amyrin synthase (AAS) to further strengthen our choice of AAS variant.

Take Aways:Dr. Cosgrove gave many recommendations for modeling and how a rigorous flux balance analysis is more applicable, and that we should also consider other alternative variants of alpha amyrin synthase.

We met again with Dr. Jiang to discuss the use of beta-cyclodextrins in the project. He had concerns of the hydrophobic nature of Ursolic Acid which might not be compatible with our current encapsulation methods. He in turn recommended multiple papers focused on different encapsulation methods, such as the use of poly(lactic-co-glycolic) acid (PLGA) as an alternative. He stated that we should consider the cost versus efficacy, and that the two might not always be sizable differences.

Take Aways:Dr. Jiang recommended we look into multiple other methods, prompting us to create a pro and con list of the different methods of encapsulation for future direction.

Agnes Slawska expressed excitement about the developments of our project. She thought that it was very interesting and impressive that we were focusing on encapsulation methods and that we had.

Dr. Magge was happy that we continued to explore Oncurex further and continued furthering our project. He was excited to continue following our project and wished us luck.

ONC1 is cancer free, but was originally diagnosed with stage 1 breast cancer 3+ years ago. Initially, she felt more calm than she thought she should have been. Her doctors had prepared her for the possibility after feeling a mass in her breast and her daughter came to all the appointments which put her more at ease. She opted for a full mastectomy because she did not want the cancer to come back and also did not want to undergo chemotherapy. She has other family members who had been diagnosed with cancer and she was scared of the thought of having to undergo that treatment. Overall, she trusted the doctors and their knowledge. She doesn't regret her decision but does wish she removed both breasts so she would not have to worry about developing cancer in her second breast. Overall, she is receptive to natural remedies. She grew up surrounded by them and would not be opposed if they were recommended by a doctor. She thinks it could be a great option but would want more information about the side effects of ursolic acid.

ONC2 was diagnosed with breast cancer at age 35+ at stage 2, and is cancer free. When diagnosed, his first thought was comparing the benefits of different treatments to the heavy toll it would take on his body. While he followed his doctor’s recommendation, he did do extra research on the side. ONC 2 received IV Chemo in addition to a mastectomy, and stated when considering his options, he wondered if he should complete his treatment in a bigger city as they might have more resources. Upon hearing our project, ONC 2 expressed excitement at the prospect of a “natural” medicine, but stated that natural does not mean good. He is in favor of using synthetic biology, and thinks “why not” as many other medicines are made using the same paradigm anyways. He believes it is too early to make an informed decision about using Ursolic Acid in general, but is highly interested in doing more research into the treatment.

ONC3 was diagnosed with stage 2 breast cancer at the age of 50+. Upon hearing the diagnosis she was afraid of dying and leaving behind her children. She was presented with the options of a mastectomy or a small reaction along with chemo. She opted for the mastectomy so she would not have to undergo chemotherapy as she did not want to become drowsy or lose her hair. She also wanted a breast reconstruction to appear the same as her past self. Overall, she was very happy with the results and is cancer free! She is a little hesitant about natural remedies and would want to ensure that the treatment she took was scientifically proved to be effective. Right now she isn't sure if she would take ursolic acid, but if more evidence came out in support of it she wouldn't be opposed if it was backed by research. She thinks some cancer patients may be interested in this type of treatment, especially those who care about where their medication is sourced from. She does believe that taking more natural medicine and creating it synthetically is good to make the medication more widely accessible.

ONC4 was diagnosed with multiple types of stage 1 breast cancer 5+ years ago and is currently cancer free. Upon learning her diagnosis, she was mainly concerned about quality of life, and how cancer treatment impacted her life and recovery. She also was concerned about body dysmorphia due to the potential mastectomy. The patient decided the best course of action for her was the one the doctor recommended, as she trusted the science due to a prior family history. Her treatment in addition to a mastectomy was IV chemotherapy, which she stated created side effects of fatigue and aversion to certain smells. Upon learning of our project, she believes the fact that it is “natural” is positive, but has never heard of synthetic biology and would be curious to learn more before creating an opinion. She stated that any research can have an impact, but her main priority is how it would impact her way of life.

ONC5 was diagnosed with stage 3 breast cancer at 75 years old. She felt a mass on her breast but waited several months before going to the doctor. By the time she went the mass was larger and already at stage 3 and metastasized. She was not optimistic about her outcome and went abroad for a second opinion because of a lack of insurance. There she searched for natural remedies with natural healers. These remedies did not work so she went to different religious shrines in hope for a miracle. After her searches she came back to the States and got a mastectomy. She also had to go several rounds of chemotherapy, with hair loss as a symptom. Overall, she is happy with the overall course of treatment. Since her cancer progressed she was not given many options but is now cancer free. She is really excited about the prospect of having more natural remedies for cancer. She searched for many natural remedies when she was diagnosed and would be excited to learn more about UA. She thinks these kinds of treatments are necessary above them to give people more choice; however, she would be worried about how synthetic extraction would affect the”naturalness” of the drug.

ONC6 was diagnosed with prostate cancer at 50+ years. He felt that his treatment options were clear to him, and we felt very comforted by his doctors. One part of treatment that he found confusing was not understanding the science behind his treatments. So, he explained that he did a lot of research on the treatments that he was undergoing to understand what they were doing to his body. He explained that when he thinks of “natural” medicine, cancer does not come to mind as the best illness to cure. He claimed that he enjoys more natural remedies for smaller illnesses, but that he had some hesitation about using something natural to combat cancer. However, he said he is very open to new technologies and is interested to see the impact that ursolic acid could have. He also expressed interest in our project and explained that he finds GMOs and synthetic biology extremely interesting and is excited about the new possibilities that they can bring to the world.

After performing a literature review, we decided to focus on ursolic acid for its unique rising potential in the field of oncology and novel method of tackling breast cancer, and chose to focus on its extraction because of the current synthesis inefficiency. We wanted to prioritize environmentally conscious methods of extracting ursolic acid and learn more about its potential as a natural treatment for cancer versus the current drugs. We also wanted to focus on cancer, instead of other potential applications like diabetes, because of the promise of ursolic acid against major cancers such as breast cancer.

Our initial goals had to be adjusted based on the actual design and implementation processes. One interview that was integral in solidifying our project was the conversations with Dr. Abbasov. Through his immense knowledge in chemical biology, Dr. Abbasov walked us through the processes and concerns connected to making small molecule drugs. He thought that Oncurex was an excellent candidate for the use of synthetic biology due to its complex pathway and process of being created. He also suggested that since the enzymatic process did not have any toxic intermediates it would make sense to focus on creating Ursolic Acid through a lens of synthetic biology.

After this conclusion we still continue to weigh the difficulties of our project as we progressed through the season. At first, we wanted to see how the ursolic acid we made would be different from that currently available. We became a little concerned about how we could differentiate our product while making sure our acid was still useful and applicable. This led us to interview Dr. Aderibigbe, an ursolic acid researcher, during the summer months. She explained to us that we could make our product even more distinct by having a specific target or adding a modification to our product. We looked into both modifications and nanoparticle carriers as potential outlets for both.

After analyzing both prospective options for modification versus nanoparticle carriers, subsequent interviews led us to decide on nanoparticle carriers as our main focal point. A conversation with Dr. Bruce Ganem pointed out modifications of UA were relatively standard in the industry as well as not environmentally friendly, which is the basis of our project. This discussion eliminated the potential use of chemical modification of Ursolic Acid itself, allowing us to explore the use of nanoparticles more. Additional conversations with Dr. Opeoluwa Oyedeji additionally pointed out the costs of modification, and pointed us in the direction of nanoparticles. However, interviews with specialists who worked with nanoparticles in the past, such as Hannah Bogich, helped elaborate upon potential risks of using solely nanoparticles such as lack of body compatibility for target absorption. As such, we considered alternative methods such as beta-cyclodextrins sEVs and PLGA, as our medium of ursolic acid encapsulation that have the same properties of nanoparticles while addressing the issue of absorption.

We also wanted to initially potentially promote ursolic acid as an alternative to drug treatment, but based on our expert interviews, we realized that, currently, it has shown the most promise for its synergistic effects with anti-cancer drugs. Based on an interview with VP of Marketing, Agnes Slawska, we also chose to focus on a specific disease. Due to ursolic acid’s proven benefits in breast cancer treatment and the overall prevalence of breast cancer, we decided to focus the scope of our project on breast cancer treatment.

Current treatments for cancer revolve around combinations of chemotherapeutic drugs. Ursolic acid has the potential to decrease the number of drug treatments a patient has to take while having an equipotent anti-cancer effect with its anti-proliferation, anti-oxidant, and anti-inflammatory properties. We need to conduct further research to see exactly how ursolic acid and synergize with which chemotherapeutic drugs to ultimately produce what is hopefully a more effective anti-cancer treatment than either modality on its own.

Our solution involves genetically modifying yeast, which does have the potential of creating other problems. There could be unforeseen consequences with the engineering of yeast, and thus it is extremely important that we thoroughly test our process and take all precautions in the lab. A specific protocol handbook was created by our Wet Lab team which not only highlighted lab protocols, but also emphasized safety. This manual has specific instructions about the disposal of wastes and the handling of yeast to ensure all members were safe in the lab. There is a small chance that the yeast could create a harmful byproduct, such as spores, but we spoke extensively with Professor Fromme in order to mitigate the chance of this occurring. Check out our Contributions page to learn more about our protocols to stay safe!

The iGEM community has important policies to ensure that our team was safe and responsible in and out of the lab during the project development process. Two major policies include not releasing any genetically modified organisms and not testing the product on humans or having engineered organisms come into direct contact with humans. Because this project worked with yeast, it was important to strictly regulate the use of yeast in the lab. Additionally, the sterility of our bioreactor is extremely important to ensure that we are only growing yeast in our tank. To make sure to preserve the sterility of the reactor we created a manual that outlines imported procedures we adhered to to make sure our yeast solution did not become infected with other foreign cells or compounds.

The target audience for this project will be biotech companies who are interested in scaling up ursolic acid production and testing ursolic acid for its health properties. These company-related stakeholders will be the most interested and affected by the project considering that ursolic acid is still in Phase I trials.

Even though our primary focus is bio manufacturers and ursolic acid suppliers, the specific application we focused on was eventually cancer treatment down stream. It was valuable to engage with physicians and patients to hear their perspectives on the limitations and benefits of current cancer treatments and gauge their interest in an alternative treatment/supplement in the future. Ultimately, we hope that ursolic acid can have positive impacts on these groups because it can provide another treatment option for cancer. As such, speaking to cancer patients and oncologists about the application of Ursolic acid as a cancer treatment was paramount to understanding all implications behind our project. We spoke to these groups mostly about the current treatments and effects of cancer treatments and their perspective on new treatments being developed.

Competing biotechnology companies may be “negatively impacted” if the project succeeds if they are also trying to develop scale up methods for ursolic acid production. We have filed a provisional patent, which will grant our team the rights to this work for a set period of time in the next years.

If the project succeeds and ursolic acid is proven safe in clinical trials, then stakeholders/developers of current cancer treatments may be “negatively impacted” if ursolic acid is widely accepted by the public in favor of current options. However, the industry can be positively impacted through collaboration and communication between biotech companies to continuously improve and foster feedback. Specifically, we can license rights to our patent to make the production of Oncurex more widespread and allow for the further collaboration with other companies. By fostering a spirit of collaboration between ourselves and biotech companies, we can hopefully widely distribute Oncurex for our true goal of befitting patients around the world.

Since our project has taken the oncology route, it is important for us to consult both cancer patients and oncologists/doctors when we are creating Oncurex. We have been closely following the clinical trials of ursolic acid into a cancer treatment. If Ursolic Acid one day meets the requirements to be used as an anti-cancer drug, it is important for us to think of oncologists and possible patients when coming up with a delivery system of the drug.

Many current cancer treatments are hard on the body. Chemotherapy can lead to nausea and the loss of hair while other treatments like radiation can cause dysphagia and fatigue. Due to these additional side effects, some cancer patients lose parts of their identity that are meaningful to them, causing their cancer journey to be even more difficult. As we continue to build and grow Oncurex, we hope to keep patients and their needs in mind when designing a delivery mechanism for the drug. Their insight will allow us to understand what factors of certain treatments they thought were intolerable and which factors were more palatable. Doctors and oncologists will help us gain a more broad view of the different current cancer treatments that are used now. They can walk us through how they come up with a treatment plan for their patients and describe what factors of a treatment make it particularly better than another.

We also want to keep in mind the impact of our project from an ethical standpoint. For example, how is this chemical sourced? Ursolic Acid is currently found primarily in fruits such as loquats and apples. We want to speak with experts to figure out how ursolic acid is currently sourced and how environmentally impactful the current extraction methods are. Considering the environmental impact of our work and the various perspectives that would be affected helped us determine the values we would consider.

Current biotechnology companies can also be consulted so we can understand more about the market we are entering. Ursolic acid is currently cleared as a dietary supplement so this already opens a market for us no matter what happens with the cancer trials. They can also help us understand how we can transition between markets if the drug does pass clinical trials and general considerations that we should have when thinking about bringing a drug to market.

Lastly, researchers dedicated to ursolic acid can also help us prioritize important factors of the project. Ursolic acid has not only been proven to support muscle growth, it is also in the beginning stages to be investigated to combat a sleuth of issues. Many researchers are indicating that Ursolic acid could be used as a possible combatant to diabetes, heart diseases, and liver disease. By speaking with researchers, we can begin to understand the full scope of the compound.

To understand the full scope of our project, we made sure to reach out to a variety of stakeholders. All of these stakeholders brought their own feedback about the potential impacts of our project – both positive and negative – and this allowed us to take steps to fill in potential gaps and to capitalize on the strengths of our project.

One weakness which we uncovered through interviews was concern about why ursolic acid was not progressing into Phase II trials. Both Dr. Joshua Zaritsky and Dr. Susan Sadoughi explained that this lack of progression is extremely concerning because very few phase I drugs succeed through to phase IV. We wanted to look into this more, so we continued reaching out to ursolic acid researchers to find the reason that ursolic acid has not progressed into phase II. Ultimately, we reached out to ursolic acid researcher Dr. Opeoluwa Oyedeji, who told us that the main reason that there are no clinical phase II trials yet is because of the barriers from the modification cost. This finding was reassuring, as it told us that the trials are still successful and should be progressing. It also further reiterated the potential strength of our project in that beta cyclodextrin would serve as an extremely helpful method because they do not require modification to the acid.

There is also the second question of FDA approval. A few oncologists and other physicians such as Dr. Magge expressed concerns about the lack of FDA approval. Ultimately, our interview with Professor Colleen Carey revealed that a main reason why the drug has not been approved by the FDA is because it is a botanical drug. Plant-based medicine is difficult to make and get approved because to make safe drugs, you need to control every aspect, and this is difficult to do with botanical drugs because there are huge issues in quality standardization. Our method of synthetically producing ursolic acid would bypass this issue, as it would mean that ursolic acid no longer needs to be extracted from plants and the issues with quality standardization and control are gone. Thus, our project actually could have huge implications in accelerating ursolic acid to FDA approval.

Furthermore, in a preliminary interview to investigate the viability of our project to succeed in the market with Ms. Agnes Slawska – a Vice President of Marketing at biotech company Invivyd – we learned that in order to effectively market our drug, it would be best to focus on a specific disease that it targets rather than just being a broad cancer drug. This finding led us to investigate the various potential uses of ursolic acid by reaching out to doctors in various specialties such as gastroenterologists, nephrologists, and internal medicine doctors to investigate the potential of ursolic acid in their fields.

Another opportunity of the project, which we discovered through interviews, was the potential of nanocarriers to make our project even more impactful. Dr. Ran Yin, an ursolic acid researcher, explained the issues with ursolic acid absorption because it is a large, hydrophobic molecule. Dr. Aderibigbe, an ursolic acid researcher, suggested that we work to differentiate our project because there is a need to either modify ursolic acid to help it perform better or to include nanocarriers that would incorporate ursolic acid.

Through a thorough literature review, we discovered that beta cyclodextrins have similar applications to nanocarrier molecules because they serve the similar function of using encapsulation-like methods to improve the delivery of hydrophobic molecules into the bloodstream (Duchene, Ponchel). This was a change to our original plan to use liposomes as a carrier method. They differ, however, in their structure and mechanisms. A previous pharmaceutical researcher, Hannah Bogich, explained that nanocarriers are accompanied with a certain level of risk and that she had previously worked for a company that chose to not use nanocarriers due to their toxic potential. This interview, in combination with our literature review, led us to pursue using beta cyclodextrins instead of nanocarriers because they served the same purpose but with less risk.

Additional concerns that were raised in our interviews concerned the accessibility of our project due to the fact that ursolic acid is already found in nature. Professor Yang Ying, the associate director for the center of life sciences ventures and a former McKinsey consultant, expressed concern about what our target market would be. She also noted that she believed it would be difficult to patent our project because ursolic already existed. These concerns led us to further investigate our ability to patent the project, and our interview with Dr. Colleen Carey explained that we could definitely patent our project through a method of use patent. Dr. Carey also expressed that our project did serve an important role because it would allow for more standardization of the production method of ursolic acid, which would improve its chances of FDA approval.

We were able to “close the loop” between our design and what is desired through our extensive interviews. Additionally, we were able to keep stakeholders involved in our project by continuously updating them on developments. Reconnecting with stakeholders that we have previously spoken with allowed us not only to update them continuously on the status of our project, but receive input at every step of the project. Closing the loop interviews with Dr. Jiang, Dr. Magge, Mrs. Slawska and Dr. Fromme helped confirm we were on the right path with our project. Understanding and envisioning how ursolic acid can be used is also vital in closing the loop. This can also include conversation with health professionals who focus on other diseases like heart disease or diabetes. We can also begin to speak with larger companies about scaling up to understand how Oncurex can be widely used commercially.

Our human practices work is extremely instrumental in understanding and implementing all our decisions. We heavily utilized interviews to develop the technical and scientific portions of our project. For example, we spoke with Dr. Jiang, who helped us understand that a lipid nanoparticle delivery system did not make sense in the context of our project. Dr. DeLisa recommended either continuous cultivation of yeast or fed batch growth along with a simple tank CSTR. This information helped our team understand how to set up the bioreactor to ensure that it can be accomplished. Dr. Specht, furthermore, gave us several recommendations on how to tweak our Gibson assemblies to maximize efficiency. He also introduced us to several methods of inserting plasmids into yeast, which allowed us to understand the most efficient way to introduce the plasmid into our yeast cell was directly because yeast already has an established pathway.

Thinking about safety and communication, it is important for us to ensure that our product does what it says it will. If we are using the ursolic acid we produce as a drug it must be in its pure form and we must ensure that it is usable medically. Dr. LuAnn Van Campen, a bioethics specialist, expressed that we should consider using whichever method allows for the most people to have accessibility and that we should consider different regions of the world differently.

It was extremely important for us to understand the perspective of physicians and patients as well. Since the goal of this project was to focus on the application of Ursolic Acid specifically in cancer, we decided to interview various oncologists that specialized in different fields of cancer. Through these interviews with the specific stakeholders, like Dr. Hyun, Dr. Saxena, and Dr. Magge, we learned about current treatments available to cancer patients and how oncologists choose treatment plans depending on the type of cancer the patient exhibits. By hearing more about the side effects of these treatments, we were able to understand ways that we can adjust Oncurex so that Ursolic Acid becomes an approved drug that is more effective. Our aim from these interviews was to reduce the risk that cancer patients would face if they took this drug, while increasing the benefit of Oncurex’s anticancer properties.

Another important factor of Oncurex is making sure it is truly sustainable. We do not want to falsely greenwash a product to make it seem more sustainable if it is not. We did this through extensive feedback and communication as well. Working with the Business subteam, we analyzed the environmental factor of our project, aiming to understand how every modification to our work might impact not only our environmental factor, but the larger context of our project. We spoke with Sophia Openshaw, an operations associate with experience calculating the environmental impact of sustainable alternatives about the calculations behind the environmental factor. She reassured us that our methodology is much more sustainable that current technologies on the market. Dr. Aderibigbe, an ursolic acid researcher, suggested that we work to differentiate our project because there is a need to either modify ursolic acid to help it perform better or to include nanocarriers that would incorporate ursolic acid. After considering both pathways, we learned through additional conversations with Dr. Ganem and Dr. Oyedeji that modifications are both not environmentally friendly and difficult to implement, leading us to consider nanoparticles more effectively.

We made an active effort to host a variety of outreach events, from tabling at farmer’s markets to teaching elementary schoolers to hosting a discussion at a local senior center. The engagement we gained with various communities such as Longview and Kendal versus Sciencenter from these events led us to approach our outreach and communication differently because each demographic and conversation taught us more about potential gaps in our project.

Our work on Oncurex was conducted under Cornell’s project team safety guidelines. First, all new members on the team received training on how to properly navigate team spaces, use PPE when necessary, report incidents, etc. as part of an online safety training course through the Cornell College of Engineering. Wet Lab and Product Development team members received additional training as necessary. To prepare for working with yeast, we spoke with Cornell Professor Chris Fromme and received a lab manual from him on how to interact with yeast safely. To prepare for working in the Experiential Learning Lab, a shared project team space in the College of Engineering, Product Development team members underwent training to understand how to handle tools and materials safely. Our whole team is dedicated to making sure our projects are safe for everyone involved. We want to make sure that our members stay safe, while also making sure the users of our product are safe while interacting with our new project.

We made it a priority to consider the ethical implications of our project by interviewing a bioethics expert and examining every part of our project to ensure that we were making ethically sound decisions. From the interview, we learned more about the importance of drug delivery and accessibility. This inspired us to research drug delivery and investigate the cost effectiveness and feasibility of each method. Overall, our project follows ethical guidelines in using yeast, a widely used model organism in synthetic biology. It does not introduce harm into the world, and the ursolic acid produced will be directed for applications in biomedical cancer research.

Overall, the goal of Oncurex is to produce medical grade ursolic acid in a more efficient and environmentally friendly way than what is currently being done. This compound shows great promise in cancer treatment, as we have learned through our literature reviews and interviews with scientific experts, so our project would be “good for the world” because it makes Ursolic Acid more accessible in a form conducive for medical application. Additionally, our project seeks to synthesize ursolic acid in a way that is less environmentally costly than current production methods. Cancer is one of the leading causes of death worldwide, making the production and research of a ursolic acid supplement all the more important to pursue.

AREA starts out with anticipation, which highlights the importance of finding a problem in our world and solving it using our project. Additionally, it requires the project to have fluidity and to change depending on new things we discover and new ideas we have.

We designed a project with multiple social impacts that touches on sustainability, agriculture, oncology, and biomanufacturing. When we spoke with Dr. Blessing and Dr. Oyedeji they explained to us that the reason why scientists are not able to use Ursolic Acid is because of its low solubility and resource consuming extraction process. So, we decided to integrate novelty into our project by adding beta-cyclodextrins to help with the solubility of the acid while also adding nanoparticle carries to highlight its possible cancer therapeutic properties.

Reflection allows for time to evaluate the current state of the project and all changes that have been made. It is important to refuse the main point of the project and to enable adjustments.

We were able to complete this through many interviews and even followed up by “closing the loop” with various stakeholders. For example, interviews conducted with Dr. Jiang allowed us to better understand the use of extracellular vesicles and nanoparticles. We had many misconceptions of some of these different particles and during this meeting Dr. Jiang helped walk us through the main differences between these different modifications. After this meeting, we were able to change and develop our initial ideas to make sure the specific carries we were using were actually complementary to ursolic acid. Further along in the season we were able to meet with him again to continue going through the specifics of these new adaptations.

After each outreach event, we planned necessary adjustments. For example, during many of our interactions with children, we noticed some language barrier. This gave us the idea to translate our children's book into various languages to allow it to become more accessible. Changes like these were able to be done by the emphasis of reflection. Additionally, we realized that children liked interactive activities. This led us to incorporate Orbeez and slime into our activities.

Engagement highlights connections made with stakeholders in the field and community members. We were able to interview people of many backgrounds like researchers, biomanufactures, ethics experts, agriculturists, cancer patients, and doctors. This year we were able to bring high engagement to Oncurex through education and outreach. We attended various events in our community like the farmers market, SPLASH, Science Center, retirement homes, and MakerFaire to engage with people of all ages. We also did events online to make them more accessible for people. We created science experiment videos related to our project, a children's book, and various handbooks.

Act is vital in making sure we are hearing the opinions of community members and stakeholders and including themes into our project as it develops further. This allows for feedback to be smoothly integrated back into the project to make use of the other points of this specific ethics framework. We were able to act by putting our observations and the advice we were given into action. We were able to edit our plan for the project and specific events we hosted in this action step due to the other steps of the framework.

Ethical means to consider how the design and development aligns with moral principles, societal values, and the overall positive impact to all those involved. This includes being conscious of safety procedures, environmental impact, and taking efforts to mitigate risks. Overall, it means aiming to avoid harm and to benefit society. We took ethical considerations of our project by engaging with a variety of stakeholders to hear their opinions on our project. This engagement allows us to mold our project so that it has the most benefit for the greatest number of people. This is because, by combining the expertise from various researchers with the lived experiences of patients, we can find out how ursolic acid can be most effective. We also practiced the ethical aspect by following lab safety procedures and doing thorough research on compounds that we are dealing with.

Legal refers to adhering to all laws, regulations, and policies which are related to research. Firstly, we adhered to the lab safety guidelines given by Cornell for the Education Learning Lab because all members of our Wet Lab and Product Development teams were given extensive safety training. Additionally, when speaking to cancer patients, we made sure to get all of our questions approved through the Institutional Review Board at Cornell. We also created an interview consent form for all of those who we interviewed to ensure that they understood that the findings from our interviews with them would be used to help shape our project.

One of the main reasons for undertaking Oncurex was the positive impact it will have on the environment. Currently, it takes a massive amount of land and water resources to harvest ursolic acid from plants. By using the method of synthetic biology we are alleviating the carbon footprint and waste created by this current process. This is allowing us to be more environmentally conscious and focused.

We also thought it would be beneficial in preparing the drug for the possibility of being a cancer therapeutic. We hope that if ursolic acid passes clinical trials it will be able to be a therapeutic that can help alleviate current side effects of current cancer treatments or be a drug with a fewer amount of side effects. This will help improve the quality of life with many people undergoing cancer treatments.

Finally, through education we were able to leave a social impact on our community. Throughout a lot of our outreach programming we had conversations with people from all age groups about the importance of synthetic biology in our society. These conversations opened many kids up to the possibility of pursuing science in the future, leaving a lasting impact in our world.

Empathizing deals with hearing stakeholders issues and challenges. One important aspect of empathizing is to listen to all perspectives without judgment.

In the context of ONCUREX, empathizing meant speaking to a wide variety of stakeholders. We reached out to doctors of various specialties, ethics specialists, ursolic acid researchers, agriculturists, cancer patients, pharmaceutical experts, and consultants to gain insight into their thoughts on the impact and feasibility of our project. A main struggle with our project was that we needed to take into account and hear how both doctors/researchers and patients thought about our project. We also hosted outreach with varying groups of people ranging from preschool age to senior citizens to ensure that we were hearing a great deal of feedback on our project and our presentation of it.

Understanding means to take these perspectives into consideration. Within understanding, we have highlighted a subcategory called “ethics standards” that involves using all of the collected perspectives, finding overlap between feedback, and handling the feedback that can be addressed feasibly. An important component of understanding is beneficence, or an obligation to handle and evaluate the risks and benefits of a project.

All of the interviews that we conducted led to a deeper understanding of ursolic acid and the issues which our project addresses. Below, we have listed a string of interviews that led us to make a change to our project:

1. Dr. Ran Yin stated that ursolic acid has poor absorption and membrane penetration.
2. Dr. Blessing A. Aderibigbe expressed that she did not have difficulty in acquiring ursolic acid, but rather that incorporating nanocarriers or modifications to ursolic acid would be extremely helpful.
3. Ms. Hannah Bogich, a pharmaceutical researcher, explained the potential risks of nanocarriers and how they have the potential to be toxic or enter different parts of the body.

Develop involves creating the technical components of the project. An important aspect to consider in this stage is non-malevolence, which means to take all precautions to avoid doing harm.

For our project, development not only meant making yeast produce ursolic acid, but also developing modifications based on stakeholder input. For example, based on the input from Dr. Yin, Dr. Aderibigbe, and Ms. Bogich, we decided to develop our ideas by doing more research on nanocarriers and other alternatives to nanocarriers.

Implementing and assessing our project means to implement the feasible and helpful changes that were recommended by stakeholders and to determine the impact of these changes and our project. An important consideration of implementation is respect for persons and justice, as it is integral to ensure that like cases are treated alike, that those participating in research are being treated with respect for their own body, and that treatment is accessible to all those who require it.

An example of implementation in our project is our addition of beta cyclodextrins to our project to improve the delivery of the drug. Based on our step develop, where we researched nanocarriers and nanocarrier-adjacent molecules, we found that beta cyclodextrins serve a similar function to nanocarriers while being more feasible within our timeframe and posing less risk. Thus, we implemented them into our project to create increased efficacy.

We collabed with McGill iGEM on their annual BIOME book with the theme of RNA or DNA-related proteins! As our project deals with the synthesis of Ursolic Acid and one component of our project involves plasmid modification, we found it apt to include one restriction enzyme we use, XBaI and how it relates to the overall design of our project!

As part of our outreach and in conjunction with our game design, Cornell iGEM collaborated with McMaster iGEM in the creation of an iGEM sticker book. Cornell created its own cute little mascots based on the fruits that contain Ursolic Acid! We developed stickers based on the mascots, and explained the use of these fruits in Ursolic Acid synthesis. We hope that these stickers represent not only Oncurex, but the playful essence of Cornell iGEM and the iGEM community as a whole!