Integrated Human Practices
For our team iGEM Toulouse 2023, it was crucial to engage in a project that could actually make a difference for everyone. We built it upon two primary objectives, i.e treating cancer with a non-GMO solution and minimizing side effects. To create a project responsible and good for the world, we needed to engage with a broad spectrum of stakeholders, well beyond scientists. That's why, in addition to meeting with oncologists and scientific experts to help us improve our design and its Modeling, we also met with supportive care professionals, volunteers from cancer-fighting associations, and patients. As outlined on our Entrepreneurship page, our aim was to implement our project in the real world, which led us to engage in discussions with entrepreneurs, specialists in clinical trials and ethics, as well as intellectual property experts. Consequently, these Integrated Human Practices have shaped our project from its conception to its integration into the real world.
Our team was motivated to participate in the iGEM competition, not only to compete but primarily to develop a meaningful project aimed at addressing a persistent and serious problem. After brainstorming ideas, our chosen cause was the fight against cancer. We reached a consensus that cancer affects each and every one of us, directly or indirectly. We all have personal connections to friends and relatives who have been affected by this terrible disease and felt powerless with this challenging situation.
“When you learn that you have cancer, everything collapses, all the projects become impossible."
told us Séverine Burdin, a former patient affected by breast cancer.
Moved by such testimonials, we were determined to take our project as far as possible. We understood that as future scientists, once we have an innovative project, it's crucial to step out of the lab to ensure its real-world impact. That's why we engaged with various stakeholders who are deeply invested in the fight against cancer. Our discussions included scientific experts in the field of cancer, clinical trial coordinators, experts in structural modeling, healthcare professionals, volunteers in cancer-fighting associations, entrepreneurship experts and entrepreneurs, ethic specialists, intellectual property experts, and most importantly, patients.
Our search for stakeholders was facilitated by the presence of the Oncopole in Toulouse, a major oncology campus hosting the IUCT-Oncopole (University Cancer Institute of Toulouse) dedicated to patient care, research and training in oncology, and the Toulouse Cancer Research Center (CRCT). Additionally, numerous support associations like “La Ligue contre le Cancer” (League Against Cancer) are available there to assist patients. This proximity to the Oncopole offered us a unique opportunity to collaborate and engage in productive discussions with an array of experts, healthcare professionals, and patients, greatly enhancing our project.
In a nutshell, our IHP approach consisted of identifying key questions requiring external advice, searching for the most appropriate people to answer them, and integrating these elements into our project. We thus divided this IHP section in nine questions:
Figure 1: Our Integrated Human Practices in nine questions.
Answering all these questions led us to build a human-centered project.
Many different treatments are used to treat cancer. It was important for us to understand their mechanisms and their limits in order to develop an innovative project that can surpass these boundaries.
How did we find answers?
We first initiated a comprehensive literature review. Many reputable organizations' websites provide precise information about the various types of treatments, such as the “Ligue contre le Cancer” (League against Cancer) [1] or “La Fondation contre le cancer” (the Foundation against Cancer) [2].
Furthermore, to ensure the accuracy of the information we had gathered, we arranged a meeting with an oncologist at IUCT-Oncopole, Dr. Ilfad Blazevic. His contact information was provided by one of our advisors. Dr. Blazevic also provided us with one of his lecture materials that present the different treatment modalities.
What did we learn?
Figure 2: Meeting with Dr Ilfad Blazevic.
The treatments for cancer generally include radiotherapy, surgery, immunotherapy, chemotherapy, hormonal therapy, and targeted therapy, as indicated in our online research. All these treatments are described as associated with side effects. Dr. Blazevic (Fig. 2) confirmed that the primary challenge with chemotherapy (i.e, the most common treatment against cancer) is its lack of specificity. It is known to affect various tissues throughout the body relatively quickly. Common side effects include fatigue, nausea, pain, hair loss, increased sensitivity to treatments and allergies, as well as a decrease in the quantity of white and red blood cells. We learnt about the different types of molecules used in chemotherapy and their associated side effects. Finally, further research revealed that chemotherapy can be administered orally, intramuscularly, or subcutaneously. However, the majority of treatments are administered intravenously.
How did it impact our project?
This literature review and the meeting with Dr. Blazevic provided us with a more precise understanding of cancer treatments. What particularly drew our attention were the numerous side effects caused by these treatments, especially chemotherapy. It is evident that minimizing side effects is a top priority and has to be taken into account for the design of our project.
While designing our system, it was imperative for us to gain a closer perspective on the actual experiences of the stakeholders involved to fully understand the reality for patients in this journey.
How did we find answers?
We first proceeded with the main institution, the IUCT-Oncopole, to encounter not only oncologists, health executives, nurses but also supportive care professionals. That is how we came into contact with Carine Vildary-Castelli (Health Executive/ Day Hospital), Alexandra Lambert and Kristell Thomas (Advanced Practice Nurses), Anne Gauci (Manager of the Supportive Care Department), Aurélie Robello (Coordinator of the UTEP- Transversal Unit for Therapeutic Patient Education -), Lisa Barthie (Socio-aesthetician), Muriel Richl (Dietitian Healthcare manager), Agnes Ruscassie (Clinical psychologist). To uncover the facilities outside the hospital premises, we engaged in conversations with members of associations that provide patient care: Joëlle Heux (Ligue contre le cancer/ League against cancer), Ludivine and Catherine (Étincelle) (Fig. 3), Marc Dupeyron (Accompagnement en Soins Palliatifs-Être là/ Palliative Care Support-Being There) (Fig. 4). Then, after becoming more proficient in the subject and improving our communication skills, we engaged with voluntary patients to provide them a platform to express their opinion. Consult our Education page to learn more about it.
Figure 3: Meeting with Catherine and Ludivine from the association Etincelle Occitanie.
Figure 4: Meeting with Marc Dupeyron from the association ASP-Être là.
What did we learn?
By interacting with these stakeholders we acquired a broad perspective of the cancer patient’s journey which helped us to better understand the need and how our project might address existing deficiencies. While it is acknowledged that each patient's experience is unique, we can discern shared steps within their respective journeys like the diagnosis announcement, the treatment and the monitoring.
What emerged from our discussion were two important points:
Figure 5: Socio-aesthetic care room at the Etincelle Occitanie House in Toulouse.
- - First, a lot of patients are suffering from side effects that could greatly impact their body and mind. By providing supportive care, the hospital and associations try to enhance the patient’s overall experience and bring them some relief by responding to their needs as effectively as possible. For example IUCT-Oncopole and the association “Etincelle Occitanie” suggest socio-aesthetics (Fig. 5), psychology or nutrition sessions to patients and their relatives.
- - Second, after consultations with medical staff and patients themselves, it appeared that there is a need for comprehensive information and education. Patients are overflowed by information from various healthcare professionals, but they lack fundamental scientific knowledge on their disease to fully understand them which could lead to distress and incertitude.
More information for future iGEMers on the cancer patient journey and the supportive care are available here.
How did it impact our project?
We had not imagined when we started how side effects of chemotherapy are a real challenge in the life of numerous patients. In addition to having to accept their diagnosis, they have to go through treatments that weaken them. The specific response of our solution to cancer cells is thus its most important feature: to significantly improve the quality of life of patients all over the world.
Moreover, our project is also an occasion to address the need for information by organizing workshops with patients to share fundamental knowledge about biology and their disease. By providing them with the choice to learn about their disease and treatment, we aim to empower patients to become active participants in their treatment plans.
Now that we have concluded that minimizing side effects is our primary objective and that these side effects arise from the lack of treatment specificity, we were keen to explore how we could enhance the treatment's specificity.
How did we find answers?
Once again, we reviewed the lecture materials provided by Dr. Blazevic, paying particular attention to the section on targeted therapies. In our regular meetings with our advisors, we were introduced to new therapies involving liposomes, which prompted us to explore scientific reviews that covered these therapies. Additionally, we had the opportunity to meet with Pr. Enrico Mastrobattista (Fig. 6), a researcher at Utrecht University specialized in the preparation and characterization of nanoparticles for the delivery of nucleic acids and proteins into cells. He shared with us his experience about working with therapeutic liposomes.
Figure 6: Meeting with Pr. Enrico Mastrobattista.
What did we learn?
We learned that monoclonal antibodies are utilized in targeted therapies to inhibit tumor growth. On the other hand, liposomes are employed as a drug delivery system, encapsulating anticancer drugs and targeting cancer cells. For instance, a ligand-functionalized HER2-targeted liposome is used for the delivery of Doxorubicin (see Description page). This approach proved to be more efficient regarding tumor reduction and more specific compared to conventional chemotherapy.
Ethically, it was not acceptable to administer GMOs to patients. Consequently, the choice of liposomes as our chassis seemed an obvious choice. Pr Enrico Mastrobattista informed us that the primary challenge with therapeutic liposomes is that only a small percentage of them reach the tumor due to the destruction of therapeutic molecules by the liver. He recommended that we enhance the composition of our liposomes with Polyethylene glycol (PEG)-conjugated lipids, as they increase the liposome's lifespan in the body by protecting them from the immune system.
How did it impact our project?
These answers guided us in creating the initial draft of our therapeutic project: utilizing liposomes for a more precise delivery of anticancer drugs to improve chemotherapy. We opted to incorporate anti-HER2 nanobodies and folate onto our liposomes to enhance their recognition of cancer cells (see Design page).
However, current liposome-based therapies have their limitations, as their phospholipid membrane can become permeable, allowing the encapsulated drug to leak out before reaching the targeted cells. This can result in side effects. And worse of all, liposomes are nothing of a classic workhorse in synthetic biology meaning we were engaging ourselves in a very difficult path.
While designing our system, it was imperative for us to gain a closer perspective on the actual experiences of the stakeholders involved to fully understand the reality for patients in this journey.
How did we find answers?
Our project's design was influenced by the thesis of Alicia Soler Cantón, titled “Synthetic Biology Meets Liposome-Based Drug Delivery”. We consulted with several experts to select the appropriate molecules to challenge our system, including Leïla Dumas (PhD graduate in oncology and a former Toulouse iGEMer), Dr. Jean-Pascal Capp (a teacher-researcher at INSA), Dr. Blazevic as well as Ben Allal and Régis Soulès (respectively researcher/hospital practitioner and engineer at CRCT). We looked on the CRCT website to find these contacts.
What did we learn?
We first learnt about some cancer cell specific hallmarks. Jean-Pascal Capp provided us with very useful information about specific oncometabolites (Fig. 7). Dr. Blazevic suggested some receptors to anchor our liposomes and advised against using diffusible antigens. Ben Allal and Régis Soulès pointed out that some receptors more present on the surface of cancer cells are also present on healthy cells (Fig. 7). For the anticancer molecules, they warned us about the possible diffusion of some drugs out of the liposome and acknowledged our idea to produce the prodrug after anchoring to avoid premature release.
From the thesis, we learned that it was possible to express a gene encapsulated inside a liposome to produce therapeutic drugs. Further research led us to discover PURE system, a solution used for cell-free expression compatible with liposome encapsulation.
Figure 7: Meeting with Jean-Pascal Capp (on the left) and with Ben Allal and Régis Soulès (on the right).
How did it impact our project?
Initially, we aimed to identify an anticancer drug-producing protein allosterically activated by cancer cells. However, this approach proved to be very challenging since engineering an enzyme so that it produces the toxic compound in the vicinity of the tumor is a very risky solution. From our discussion with the experts, we decided to focus on something more generic with the production within the liposome of the enzyme capable of converting the inactive prodrug into an active anticancer drug. Our selection of the enzyme thymidine phosphorylase was based on the literature.
To control this production, we decided to couple this system with a genetic biosensor inside the liposome. However, the experts warned us about the difficulty to have a truly specific response so we chose to develop not one but two biosensing systems to increase our chance for specificity: one that utilizes the split T7 RNA polymerase, and the other based on the 2-HydroxyGlutarate control of the repressor DhdR (See Design). Finally, we chose the anticancer 5-FU compound for our proof of concept as it is one of the most used drugs, and Tegafur as its direct non-toxic precursor.
In conclusion, we have conceived an innovative approach to mitigate side effects through synthetic biology: encapsulating a transcription-translation machinery coupled with a genetic biosensor inside the liposome to initiate the production of the anticancer drug directly at the tumor site.
Before starting experimentation, it was essential for us to know if our newly-designed solution was realistic. We also needed to foresee how it could be used with patients.
How did we find answers?
We consulted with several experts including Ben Allal (researcher/hospital practitioner at CRCT), Régis Soulès (engineer at CRCT), Dr. Thomas Esquerré (anesthetist at Toulouse CHU) whose contact was given by one of our advisors and Pr. Enrico Mastrobattista.
What did we learn?
Dr. Thomas Esquerré explained to us that our solution could be of interest in both surgical procedures to eliminate the remaining tumor cells or in injection (Fig. 8). He was enthusiastic about the versatility of the treatment. Ben Allal and Régis Soulès pointed out that the size of our liposomes could be too large for intravenous injection. This remains a challenge of our project but we wanted to find avenues of exploration on how to reduce their size. Jois et al. in his article [3] affirmed that “nanoparticles with a diameter between 20 and 200 nm are suitable for drug delivery to cancer cells”. We discussed the feasibility of size reduction with Pr. Enrico Mastrobattista. He was also familiar with previous research conducted by Pier Luigi Luisi, which was related to protein synthesis within small liposomes. In his work presented in his article [4], Pier Luigi Luisi successfully achieved the expression of GFP in 200 nm liposomes.
Figure 8: Meeting with Thomas Esquerré.
How did it impact our project?
We greatly appreciated the positive feedback we had. This convinced us of the suitability of the method and its usefulness in many situations for many patients. Even if some technical aspects will have to be addressed such as the production of calibrated small liposomes, our design was very promising so we went on the next step.
As the design of our project was quite challenging, we wanted to ensure its feasibility in silico. A first question was to know if the split T7 can be functionalized for HER2 recognition (see In silico protein design page). Another question was about the yield of 5-FU production and its adequacy to treat cancer cells.
How did we find answers?
To answer these questions, we designed a global kinetic model and a structural model. Our global kinetic model was constructed using COPASI [5] and by incorporating some work from iGEM team Duke 2021 [6] and iGEM team Delft 2021 [7].
To develop our structural model, we consulted with many experts: Philippe Urban (scientist at TBI leading the Metabolic and Molecular Engineering team), Sophie Barbe (research-director at TBI), Juan Cortés (research-director at LAAS-CNRS and team leader), Younes Bouchiba (PhD student at TBI), and Ilinka Clerc (PhD student from Juan’s team) (Fig. 9).
Figure 9: Meeting with Juan Cortés.
What did we learn?
First, Philippe Urban, a regular user of the structure predictor AlphaFold2, explained to us how the algorithm works, how to use it and how to interpret the results. We met several times so he could follow our progress and help us when we had some issues. Then, we exchanged with Sophie Barbe, Juan Cortés and Younes Bouchiba who work on structural bioinformatics and protein engineering. We held regular meetings with the three of them to figure out a strategy for optimizing the transmembrane linker of the split T7 RNA polymerase biosensor.
Once the strategy was found, Juan Cortés and Ilinka Clerc provided us with mathematical tools and algorithms from LAAS-CNRS to better study the disordered regions of the transmembrane linker. They taught us how a statistical study could help us and analyzed every result with us and Younes Bouchiba. These experts helped us to obtain a successful model of a split T7 RNA polymerase using analysis methods easily accessible to other teams. This will increase the success rate of producing our activating enzyme.
Second, our global kinetic model showed that the synthetic liposomes may produce 5-FU at a concentration dependent on the surrounding environment, hence demonstrating the feasibility of our project for targeted chemotherapy.
How did it impact our project?
This information was essential to us. Without it, we would not have engaged in the split T7 RNA polymerase approach as it was the riskiest challenge. Likewise, we made a big effort on the entrepreneurship part because we knew that the system was capable of producing enough anticancer molecules to be actually used.
Entrepreneurship is crucial to ensure the accessibility of our treatment to patients and to bring our project out of the realm of theory.
How did we find answers?
Figure 10: Meeting with Marc Dubourdeau.
Figure 11: Meeting with Julien Durand.
Figure 12: Meeting with Yves Dudal.
We met with numerous individuals associated with entrepreneurship, ranging from startup incubators (Le Catalyseur, Pierre Potier Center) to entrepreneurs (Julien Durand, co-founder of Sweetech, Yves Dudal, CEO of Perseo Pharma, and Marc Dubourdeau, CEO of Ambiotis) (Fig. 10, 11 and 12). We also engaged with entrepreneurship stakeholders (Michel Lomi and Pierre-Alain Hoffman, both professors at INSA, and Elvire Prochilo, a consultant in management and entrepreneurship). Additionally, we focused on intellectual property by meeting with Philippe Lucas from INPI (Institut national de la propriété industrielle) and a business manager at TTT (Toulouse Tech Transfer). Finally, we consulted with Thomas Dubrunfaut, a clinical trials coordinator. We initiated contact with these people by emails, which we obtained from various websites or through connections from our school.
What did we learn?
Each meeting provided us with valuable insights into creating a therapeutic startup and developing the best strategy. Elvire Prochilo played a significant role in helping us to create our business model and business plan.
A significant portion of our inquiries concerned intellectual property protection, specifically the patent application process. In this context, we explored the relationship between a startup and the laboratory holding the patent for the startup's technology.
We also acquired knowledge about fundraising and learned how to create crowdfunding campaigns. We improved our pitch skills through practice during meetings and by participating in entrepreneurial events (Fig. 13). This skill is essential when persuading potential investors. Additionally, many entrepreneurs emphasized the importance of being supported by startup incubators. As a result, we reached out to the Pierre Potier Center to understand the various steps involved in establishing our startup within their premises. They also provided insights into the partnerships that startups need to cultivate to develop their activity.
Given that clinical trials are a key milestone of our future strategy, we engaged in discussions on ethics with Thomas Dubrunfaut, CRO Clinical Trials Coordinator.
Lastly, we acquired the ability to anticipate and manage risks within a startup.
Figure 13: Participation in the 2023 edition of “Le Catalyseur” Committee, with Colette Schenker.
How did it impact our project?
These many discussions led us to decide the best strategy for our future startup and establish the sound business plan described here (Entrepreneurship page).
Ultimately, our project has the potential to positively impact the lives of patients. However, it is essential that they are willing to accept this innovative treatment, which uses liposomes by injection. This is why it was important to engage in a dialogue with them, to take the time to explain our system, and to address their questions and concerns.
How did we find answers?
Figure 14: Workshop with patients members of the Etincelle Occitanie association.
In order to receive the most accurate response, we encountered individuals who have endured or are currently facing cancer. They agreed to discuss with our team under the condition that we preserve their anonymity. We were able to establish a climate of trust during our workshop on biology within the Etincelle Occitanie association (See Education) (Fig. 14). Outside the workshop, we had the opportunity to converse with Séverine Burdin which also gave us an insight on the perception of treatments by patients.
What did we learn?
Our system was well received as we explained that liposomes were biocompatible, and would not trigger the human immune system. Discussing our project after our workshop on biology and cancer was very useful as they could better grasp the functioning of our system. The receptors on the surface of our liposomes for specific anchoring, along with the local production of the anticancer drug, ensure a better specificity that inspires hope in them. In fact, they were enthusiastic upon learning about our strategy, which holds the potential to offer future patients a higher quality of life and improved effectiveness. Some have already undergone innovative treatments, and the efficiency of these treatments was greatly appreciated despite side effects.
Even without knowledge of fundamental biology, former patients like Séverine Burdin also reacted favorably to our system as "[they] have to trust, there is no time for thinking” when you learn about your diagnosis. As we already mentioned, information can be overwhelming at the beginning of a treatment, and patients often choose to wait before asking more questions, especially about the functioning of their treatments. Similarly, they are accepting the intravenous mode of administration of our project as it is the only option to deliver effectively our curative liposomes.
How did it impact our project?
Our discussions gave us a good perception of our system by patients. Their primary objective is recovery, and thus the utilization of liposomes or cell-free systems is accepted as they can bring more efficacy and reduce side effects. This demonstrates the potential of our system as a compelling alternative for future patients.
As Edgar Morin, a French sociologist and philosopher, said "The more powerful general intelligence is, the greater its ability to address specific problems.”
That is why we concentrated our efforts on one problem: curing cancer while reducing side effects of chemotherapy affecting millions of patients. However a project like ours can raise ethical issues. Ethics can be defined as the questioning regarding human actions that don't necessarily lead to a single, definitive answer. To prevent any ethics violation, Béatrice Jalenques-Vigouroux, a lecturer in ethics and sciences of information and communication, advised us to clearly define our project’s values.
How did we find answers?
On the recommendation of Béatrice Jalenques-Vigouroux, we were able to meet with Pr. Bettina Courderc, who is currently working on bioethical issues in health innovations, and a member of the Personal Protection Committee (CPP) which ensures the ethical compliance of clinical trials. Our ethical approach was completed by Thomas Dubrunfaut, Clinical Trials Coordinator at a CRO. We requested the assistance of Julie Fortin and Sophie Desgranges, coordinators of the “Equal Opportunities O’talents” Program, and Sophie Yvon, researcher & science communicator. Aurélie Robello was also able to help us with her expertise in communication as a coordinator of therapeutics workshops at IUCT-Oncopole.
What did we learn?
Our objective was clear: to cure cancer patients while maintaining a good quality of life by reducing side effects of chemotherapy. From this, we defined our core values that helped us to integrate ethics all along the project.
INTEGRITY
- - As scientists, we have a responsibility to present our work accurately, enabling fellow researchers to contribute to the progress of our project and thus science.
- - Integrity is essential for Pr. Bettina Couderc, as we will be required to conduct clinical trials on our system.
- - As Thomas Dubrunfaut advised us, we need to evaluate if the benefits outweigh the risks before presenting our system to clinical trials Phase I and II.
- - Patients applying for clinical trials must have time to fully understand the conditions of the contract before agreeing.
MUTUAL EXCHANGE
- - Our meaningful exchanges helped us to better understand the daily life of patients but also of healthcare professionals, volunteers of associations and relatives.
- - By discussing with the professionals at IUCT-Oncopole, in particular Aurélie Robello, we started to discern a need for information and education as most of the patients didn’t have a scientific background and more precisely biology knowledge.
- - More exposition to these concepts from a young age could help people to understand the potential biological changes in their body during their life. That is why we thought of contacting Julie Fortin and Sophie Desgranges to learn how to adapt our speech to middle schoolers. Moreover, to ensure the best progress of the exchange, Sophie Yvon stated the importance of good preparation in particular in selecting words wisely.
SECURITY
- - As a health initiative, it is subject to rigorous regulation in accordance with Good Laboratory Practice guidelines, ensuring the highest standards of safety, quality, and ethics in our laboratory work.
How did it impact our project?
INTEGRITY
- - Throughout the numerous presentations we made, for example, during the scientific symposium BioSynSys on July 19th, we presented our results with transparency and truthfulness (See our Education page).
- - We will impose criteria in the contract drafted and approved by our expert CRO partner to exclude patients from the trials Phase I and II if they do not respond favorably during the course of the treatment (See our Implementation page).
- - The preclinical tests on cells and animals will be able to determine if our system could bring more benefits than risks. The well-being of the animals is also a priority for us (See our Entrepreneurship page).
- - Furthermore, we will ensure that every patient comprehends the trial conditions to provide informed and voluntary consent while retaining the freedom to withdraw for any reason.
MUTUAL EXCHANGE
- - The dedication of professionals and volunteers and the courage of the patients themselves were truly inspiring and motivating for us. We are committed to do everything we can to develop our solutions, as people are depending on the outcome.
- - We chose to address the lack of knowledge in biology as we were convinced it could help them in their fight. That is why we prepared a workshop to explore with them the fundamentals of biology and address the main questions they might have regarding their disease (See our Education page).
- - We also decided to engage with middle school students to introduce them to a more tangible aspect of biology. By helping them understand the relevance of biology in their lives, we hope to foster their interest and help them retain these concepts, as they could prove useful at various points (See our Education page).
SECURITY
- - We rigorously followed the Good Laboratory Practices during our first experience and will be essential in the future of our project especially in the preclinical trials (see our Safety page).
To conclude, our human-centered project has a focus: to suggest a cancer treatment that has fewer side effects, making life easier for patients. Throughout its development, we have maintained a commitment to create a positive impact in the real world, driven by the principles of responsibility and ethical conduct.
Our approach has been consultative and inclusive, involving diverse communities (scientific, medical, associations, patients) to ensure that our objectives align with genuine needs and that our core values are thoughtfully integrated into every decision we make.
Our team has done its best to highlight qualities of reflection, responsibility, and responsiveness, ensuring that we remain flexible in front of challenges, but also considerate and accountable of our actions as we work towards our objective.
In the end, our project is the reflection of our commitment to improve the lives of patients and to bring positive changes in global healthcare.
References
