Cancer is a leading cause of death worldwide, accounting for nearly 10 million deaths in 2020, or almost one in every six [1]. Despite advancements in science revealing the complex interaction of variables responsible for malignant phenotypes, successful treatments for most cancers are still a long way from reality.
Traditional cancer treatments rapidly kill growing cancer cells and normal cells. This leads to many adverse effects. Targeted drugs represent an improvement over conventional therapies but have several flaws; for example, antibody-drug conjugates often cause significant side effects; pathway inhibitor drugs rely on specific pathways, which sometimes can be bypassed by tumour cells [2].
As we learned of all these issues, we were determined to find a means to aid the millions of people suffering from cancer and its harsh treatment side effects. We began by learning how cancer medicines function and tried to get to the bottom of the issue. It was immediately apparent that present drug delivery systems were severely inadequate due to their unaffordable costs and off-site targeting. Therefore, we chose to build a more robust, affordable drug delivery system with improved specificity and efficacy.
A group of 15 enthusiastic research students formed a group to seek and tackle the challenge of off-site targeting by leveraging the ability of synthetic biology to synergistically combine different approaches in cancer therapy. We started working extensively towards this goal, and eventually, it led to the creation of Duonco, a novel dual nanovesicle drug delivery system that we believe has the potential to reduce off-site targeting. We hope to establish a new method for drug delivery that can be improved and adapted to better serve the needs of patients. To our knowledge, this is the first time such a dual nanovesicle system has been designed for targeted therapy.
Our next objective was to select a suitable model for our project. We went through the literature on current statistics of cancer rates in India and found out that breast cancer has become the most prevalent form of cancer, with the highest gross incidence and mortality. A few of our members also attended a medical camp held in our community, where they learned about the dramatic increase in breast cancer cases and their lack of knowledge of the diagnosis, causes, and treatments. This motivated us to select breast cancer as our model disease and move toward developing our drug delivery system.
As our project progressed, we engaged with oncologists, doctors, and drug delivery specialists to finalise the components of our prototype. Our project modelling proved to be incredibly useful for comprehending the dynamics of OMV internalisation and augmenting our wet lab. While speaking with non-governmental organisations (Hair for Hope, Knitted Knockers), we observed the lack of knowledge and stigmas around breast cancer. This motivated us to organise talks and a drama to raise awareness, a breast cancer screening camp, a hair donation campaign, awareness pamphlets, and other outreach events. We also conducted a survey via online and offline platforms to analyse the awareness of the general public on breast cancer and related topics. This gave us insights into people's misconceptions (males cannot get breast cancer, and breast cancer can cause pregnancy issues), and we made it a point to ensure that all of these issues were covered while preparing our outreach initiatives. Our meeting with companies and the government startup commission helped to get perspective on how to further our concept and enabled us to build a suitable plan to introduce Duonco to the public in the best manner.
When we started exploring our project more, we felt it would be best to develop a framework to execute our IHP effectively. Each step was planned in a manner that would help in working out the one that followed it. Our team used this method to ensure that discussions and lessons learned from one project stage were carried over and incorporated into the next. This made it easier for us to ensure that Duonco, as a drug delivery system, mirrored the values determined to be significant to our stakeholders.
The opinions of stakeholders, drug delivery experts, academia, cancer scientists, pharmaceutical companies, the general public, caretakers and doctors have all contributed significantly to determining the course of our project at each level of our human-centred, four-step approach to IHP.
As we began our project, we realised it was of utmost importance that we try to understand the magnitude of the problem we are addressing.
In the first stage of our approach, to gain some preliminary answers to these questions, we took several measures: Preparing a survey to analyse the public's awareness of the subject, scouring through literature to gain a broader understanding of the severity of the issue and its significance in our society and talking to social scientists. These helped us identify our major stakeholders:
In the second stage of understanding our problem, we interacted with our major stakeholders. We held interviews with breast cancer survivors (interviewed seven survivors) and doctors (four doctors) about the cost, efficacy, efficiency and affordability of current targeted drugs. We spoke with pharma companies such as Sanofi Aventis to understand the process, right from the start of funding research for medicines to each stage of trials, marketing strategies and legal procedures. We learned about the outlook of the general public towards genetic engineering and the use of genetically modified organisms (GMOs) in medicine. From our interactions with professionals in the field of science communication, we developed an effective system for synthetic biology education.
Having identified and understood the various aspects of contemporary cancer therapies, our team began brainstorming solutions.
After discussing and analysing our stakeholders' needs, we decided on a synthetic biology solution. These are all aimed at an effective mechanism to administer the medicine to tumour cells while keeping other cells safe. It was also where current methods fell short due to their high cost, lack of specificity, and failure to provide a sustained immunogenic action.
Key Points We Received From Our Stakeholders Are:
We began to go through published literature and found E.coli outer membrane vesicles (OMVs) as an attractive candidate for our nanocarrier due to studies on its efficiency as a drug carrier in in-vivo experiments. We met with several experts in the field— Dr. Denice Bay, Dr. Gujarati, and Prof. MRN Murty and decided upon OMVs and the usage of a dual OMV system to enhance the specificity of the therapeutic.
We finally chose our model to be breast cancer, realising its significance in our immediate locality from statistics and our firsthand interactions during UBA (UNNAT BHARAT ABHIYAN) medical camp. In Thiruvananthapuram, breast cancer had the highest incidence among all cancers and accounted for one-fourth of all new cancer cases in 2018-2019 [3]. All of this information cemented our desire to develop Duonco for use in breast cancer therapy.
To form a solution, we continuously met with our stakeholders and integrated their feedback throughout the design process. We strived to ensure Duonco holistically embodies the needs of everyone involved in our stakeholder chart.
We corresponded with wet lab experts and our PIs to design the experiments required to build and test our system. We prepared a list of materials and budgeted. We came up with a project plan comprising three phases:
Given the relatively short duration of the project, we chose to restrict our scope to generating engineered OMVs and testing their uptake. We attempted to supplement our experiments with results from math modelling and vice-versa. Primarily, we used mathematical modelling to obtain predictions for components or processes we were unable to experimentally achieve. This included electroporation to load the cargo into the OMVs and prodrug-enzyme interactions. We found a general correspondence between the results from math modelling and wet lab experiments, especially relating to protein expression. We parallelly planned awareness programmes and events to take our projects to people who really needed them.
Our final identified value was that the solution be financially viable and scalable, allowing realistic implementation of the solution. Synthetic biology lends itself well to being scalable and has excellent potential to be a financially viable solution. We made it a point to keep our four points from stakeholders during ideating to be the driving factor of our design for Duonco.
Having defined our problem, our project and our plans, we organised our team to effectively work towards our goals. We distributed tasks among team members. We set a timeline for our work and frequently discussed our progress with our PIs, who always provided us with feedback.
We continued to meet with experts to better understand different wet lab techniques, such as electron microscopy (to image OMVs) and dynamic light scattering (to characterise OMVs). We reached out to researchers who kindly contributed hyper-vesiculating E.coli strains, plasmid backbones and breast cancer cell lines SKBR3 and MCF-10A. We procured other materials, such as antibodies and restriction enzymes, in the form of sponsorship from companies. This set us up for fruitful experiments in the lab.
We conducted several project awareness programmes like project reveal, awareness talk, distributing pamphlets etc, which all were inspired by the survey analysis and public interviews.
Our entire project, including our IHP journey, has been focused on people, and our proposed implementation is no different. Returning to our stakeholders and specialists throughout the implementation planning phase allowed us to close the loop and make decisions based on their values and needs in terms of ethics, technology, safety, and communication. Talking to the Kerala startup mission and drug delivery companies, we were able to devise a proper future plan to scale DUONCO to its next level.
Understanding the impact of our project on the community, receiving input from the stakeholders and establishing a feedback loop is a fundamental component of integrated human practices. To facilitate this loop, we interacted with several individuals and organisations throughout the course of the project.
In 2020, the number of cancer cases rose to 19.3 million, while the number of deaths surpassed 10 million. Cancer reach is accordingly ever evolving to battle against this exponential rise in cases. Many of the existing therapies for cancer have significant side effects associated with them. In order to fully comprehend the limitations of the existing therapies, we interacted with patients and clinicians who could give us a first-hand account of their impact.
Unnat Bharat Abhiyan (UBA) is a central government initiative that enables higher education institutions to visit rural areas of India and bring in development in those areas.
On International Women's Day 2021, some of our team members volunteered for a breast cancer screening camp in association with the UBA committee of our institute at a nearby village of Chathancode, Kerala. They realised how much of an issue breast cancer has grown to become in a few years. In 2020, breast cancer became cancer with the highest gross incidence and mortality, both in India and worldwide. Even in our little community of Vithura (with a population of 20,000), the incidence of breast cancer patients was alarmingly high. This demonstrated the severity of this disease and the urgent need for a more effective treatment method. These statistics and the helplessness of the community we observed led us to choose breast cancer as a model to perform our study.
UBA Breast Cancer Awareness Medical Camp at a tribal area in Vithura
To better understand the side effects of current treatment methods, we spoke with a teacher who is a cancer survivor. Treatment was a traumatic experience, where she encountered the side effects of chemo and radiotherapy, such as hair loss, body weakness, and mastectomy-caused lymphoedema, not only affecting her overall state of well-being but also impacting her professional life, showing us the need for a better solution. She was open to the idea of using GMOs as an alternative, provided it helped alleviate some of the symptoms, showcasing to us that the general public might be receptive to synthetic biology solutions to real-world problems.
This interaction also made us aware that lymphoedema is a common condition for people who have undergone mastectomy. We decided to interact with respective stakeholders to get a better understanding of mastectomy. In addition, we decided to meet more cancer survivors and have a better understanding of the frequent issues patients confront throughout treatment to address some of them through our project.
The end users of our novel treatment method are the patients. Getting to know their difficulties and expectations is essential. Though cancer is widely spoken about, some aspects of cancer therapy, including mental and physical frailty, societal pressure, and financial concerns, are never under the spotlight. Therefore, we interviewed five breast cancer survivors. We obtained their permission prior to conducting our semi-structured interview. The consent form is attached here. Their perspectives on various breast cancer elements are described below.
| Topic | Perspective |
|---|---|
| Awareness level | People lack knowledge of early detection. Typically, cancer signs are not recognised until a later stage, making treatment challenging. Most of the interviewees shared similar experiences. When a lump developed, no medical opinion was taken. Only when it began to spread did they conduct tests. Even minor ignorance can cost a great deal.
Men, particularly, took more time to get medical assistance due to the stigma that men can't get breast cancer. Thus a delay in recognising the condition is present. |
| Side effects | Most of the participants have undergone chemotherapy, and some have undergone radiation therapy and/or immunotherapy along with chemotherapy. Many of them have undergone surgery (mastectomy) before getting any other treatment, as they detected the disease at later stages. Receiving multiple treatments has severely affected the patients. Almost all of them were physically weak and felt nauseous during the treatment period. Patients who had undergone mastectomy and required physiotherapy developed lymphedema. Hair loss was experienced by the patients after undergoing chemotherapy. Several other side effects were shared by the patients, like difficulty swallowing, low blood count, constipation, and vomiting. |
| Social life and Mental health | All the participants' lives were altered by contracting this condition. Their career, family, and social lives were all affected. During the treatment period, they were unable to work due to the chemotherapy sessions and the fatigue that followed each session. Patients who were homemakers were also affected.
The interviewees described their feelings of mental fragility throughout those eras when society regarded them differently. People treated them with sympathy, which made them uneasy. Many of them struggled financially during those times. |
| Future directions | Participants discussed treatment strategies and societal attitudes concerning cancer patients. They are willing to take drugs containing GMOs upon showcasing their safety. They want the procedure to have fewer side effects. Additionally, they desired that the new treatment avoid hair loss. In addition, they hoped that people with cancer be treated equally and without prejudice. |
Adv. Baburaj is the Vithura Grama Panchayat President. Panchayat is a local village council. The Panchayat president is an elected representative who chairs the panchayat meetings. We spoke with him to learn about the current breast cancer statistics of the Vithura Panchayat. He gave us a general idea of the situation in our locality. We also wanted to know of existing programs by the local and central governments to increase detection tests for cancer. He connected us to Asha workers. ASHA workers are Accredited Social Health Activists who are actively involved in health awareness and implementation in rural areas.
At Vithura Panchayat Office
To learn about current treatment methods other than chemotherapy, we visited KIMS Health Centre in Thiruvananthapuram, Kerala. We met Dr. Bharath V.M., a surgical oncologist at KIMS. He introduced us to Breast-conserving surgery (BCS) or lumpectomy, where only the lump with roots will be removed surgically. He pointed out that all patients cannot undergo BCS. Even the patients who have undergone BCS may need to go through other treatment methods later, like radiation therapy, hormone therapy or chemotherapy. BCS can have side effects like pain, scar, dimples, or lymphedema in the arm. We understood that a better treatment method is essential. Then he discussed the gravity of HER2-positive breast cancer. It is a particularly aggressive form of breast cancer. The stages of HER2+ breast cancer is determined using a scoring method. The degree of HER2+ protein expression on the surface of breast cancer cells is indicated by immunohistochemistry scores ranging from 0 to 3. A score of 0 to 1+ suggests that it is HER2-positive. A score of 2+ indicates that the cells are borderline, while a score of 3+ indicates that they are HER2+. This information was relevant to us as we wanted to be sure HER2 receptors could be targeted by our drug delivery system.
Dr. Bharath V.M.
We interacted with Dr. Kavitha Ravi, MD Pathology, who is an additional professor of Pathology at the Government medical college in Thiruvananthapuram, Kerala. She provided us with cancer statistics in the city we live, Thiruvananthapuram. She confirmed the need for a drug delivery system targeting HER2. It is crucial due to the aggressive nature of the disease and the chances of relapse in other organs despite current treatment methods. She highlighted the positive effects of targeted therapy in which only malignant cells are affected. She also visited our institute and delivered an awareness speech regarding breast cancer to our staff.
Dr. Kavitha Ravi.
Dr. Sreekala Sreehari, MD FRCPath, is the Clinical Director, Pathology at University Hospitals Dorset NHS Foundation Trust. Aside from her years of experience as a histopathologist, she and a few other doctors created the Brexa app. It assists people in detecting breast cancer early and directing further treatment. It is used by thousands of people all over the world. It instructs users on how to perform self-breast examinations.
In our interaction with her, she reviewed our survey questions to ensure they were relevant and accurate. She emphasised that there are other types of breast cancer besides HER2-positive ones, like triple-negative breast cancer. Despite these cancers being of high grade, they lack the receptors that our drug delivery system targets. This interaction prompted us to learn more about other types of breast cancer and potential targetable receptors. This will aid in the expansion of our project Duonco to a broader range of cancer types. She discussed the issue of breast cancer detection in India, where people do not voluntarily undergo screening tests, to be a primary problem for the high rise in cases in past years. We recognised the importance of raising awareness in our society. We planned a number of public awareness campaigns to help with early detection, specifically Self Breast Examination.
Left- Dr. Sreekala Sreehari
Right- Screenshot of the meeting with Dr. Sreekala Sreehari
Once the team decided to use a novel drug delivery system to target breast cancer, we felt it was critical to ascertain how well-informed the general public is about the disease and whether there are any myths that we can dispel. So, with the help of Dr. Harilal Madhavan, we created a questionnaire and made sure we adhered to all the safety requirements for using human subjects in research. We made our survey available online in June 2022 in 5 different languages. We also collected print surveys from people that we believed we couldn't reach through online platforms.
We received over 300 responses for our survey, and we particularly aimed to find correlations between different parameters we asked in the questionnaire.
The suvey is linked here.
The links for the survey in different regional languages- Malayalam, Bengali, Tamil and Hindi
As soon as we began defining our stakeholders, it became clear that the general public was the most significant stakeholder in our project. We, therefore, believed it was imperative to reach out to them and learn more about their perceptions of cancer and drug delivery mechanisms. Team members travelled to their home states in different parts of India—Kerala, Tamil Nadu, Maharashtra, and New Delhi— and went out and spoke with people from various walks of life to get a general sense of how issues related to breast cancer are viewed. We obtained their permission before filming the interview. Read more.
These exchanges gave us a lot of new information, including the fact that:
This served as our motivation for printing out informational pamphlets with essential breast cancer awareness information.
“Would I have to be known as Barbara who had breast cancer or would I be Barbara as I was before”
Barbara Demorest is the founder of the Knitted Knockers Support Foundation. She is also a breast cancer survivor. We interacted with her to learn more about her journey and the need for knitted knockers. She faced several difficulties during her treatment days and felt that her mental health often took a backseat to her physical well-being. Her inability to have reconstruction surgery or use a synthetic prosthetic kept her from returning to life. Read more.
Knitted Knockers was introduced to her through her doctor. Finally, she gained a sense of freedom to go out, hug people and wear a regular bra without having to use any medical prosthetic device or specialised bra. This gave her a sense of normalcy, and this was her inspiration to form KNITTED KNOCKERS. Several people who joined the initiative could relate to the shared trauma that they all faced.
Her strategy was to raise awareness, get the local community involved and provide for their region. Knitters make the knockers and send them to their headquarters, where they stuff them and post them to clinics. They have spread across all states in the USA and six other countries in a few years. Talking to her really gave us the prompt to start small and achieve big.
Left- Barbara Demorest
Right- Screenshot of the meeting with Barbara Demorest
By interacting with mastectomy survivors and Ms Barbara Demorest, we learned that those who have undergone a mastectomy confront numerous challenges. They experience discomfort and may have postural issues. During our conversation with Dr. Kavitha Ravi of Thiruvananthapuram Medical College, we learned that the Regional Cancer Centre (RCC) in Thiruvananthapuram provides post-mastectomy breast prostheses. We went there to learn more about this. Read more.
The centre is managed by nurses who are responsible for the patient's well-being. We visited Ms Geethakumari D., the head of the nursing staff. She explained that utilising a prosthesis or undergoing breast reconstruction surgery is entirely up to the patient's discretion. However, after a mastectomy, most women prefer to use one of these alternatives because their confidence has been impacted. She stated that prostheses are more popular than breast reconstruction surgery because they are less invasive. Patients who require silicone breast prostheses are provided with them. It cannot be used immediately after surgery because the area near the chest is extremely painful and may become contaminated. These prostheses are generally suggested two months post-surgery.
We learned a lot from our visit, especially how the disease changes the lives of affected people even after their treatment and how it leaves behind issues of body image. We also made a note to provide information about the options available to patients post-surgery available in our future interactions with people.
At Regional Cancer Centre, Thiruvananthapuram
Summary
Our interactions with these doctors and cancer survivors reminded us that a more effective treatment strategy is necessary. It has to utilise the existing therapy medication, which is highly beneficial and reduces significant side effects. We chose to design a delivery system that uses not one but two cancer cell-specific cell surface receptors to improve the specificity of drug delivery and mitigate the side effects caused due to offsite targeting.
The central theme for the design of our project was to develop a drug delivery system that would enhance the specificity of the chemotherapeutic agent. Thereby, it would circumvent most problems associated with off-site targeting, which is significantly high in the case of traditional chemotherapy. We had quite a few queries regarding the design of the project. They ranged from receptor targets of our tumour-homing peptide to defining models that would supplement our wet lab work. Thus, we interacted with experts in the field to receive their feedback and build upon our initial idea to develop a more robust project design.
Prof. Mathew is a professor at the National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore. In addition, he serves as a visiting professor at IISER Thiruvananthapuram.
We initially met him during the infant stages of the project, when we intended to develop the drug delivery system using only one type of vesicle. He pointed out to us that it would not be very different from the existing targeted therapies and encouraged us to think about adding a double-check mechanism. During our second interaction with him, he assisted us with our mathematical modelling. He asked us to determine the drug concentration necessary to kill cancerous cells. This will assist us in quantitatively estimating the amount of prodrug/enzyme that needs to be loaded into the OMVs and, thereby, the pore size required to maintain the integrity of the membrane. We had planned to use a hydrophilic dye for showcasing internalisation. He suggested employing lipophilic dye. We realised that the use of lipophilic dye was a superior alternative to the use of hydrophilic dye. But due to limitations in resources, we could not employ it in our lab work, but we hope to incorporate it in our future work.
Prof. M.K. Mathew
Dr. Gujrati is Research Scientist and Consultant at the Institute for Biological and Medical Imaging, TUM Germany. He has excellent experience in various fields like drug discovery, non-clinical pharmacology, bioimaging, scientific writing, and immune and regenerative therapy.
Through this meeting we got several insights regarding subcloning and construct design. While designing our constructs the length of the linker is not significant, the order of linker and tag can be interchanged. Dr. Gujrati also kindly agreed to provide us with pGEX-4T1-ClyA_Myc_Affi, this served as a base for our entire cloning process. We had a few concerns regarding the stability of proteins post electroporation, he assured us that it would not be an issue and mentioned that while selecting our prodrug enzyme system we would need to take into account the size of the pore created via electroporation. As we were unsure if PBS would act as a cryopreservant he clarified that the use of PBS and other saline solutions would be the ideal buffer to resuspend our OMVs. He also cautioned us to maintain a sterile environment and use vacuum filter pumps to avoid contamination. Overall, his work on bioengineered bacterial outer membrane vesicles as cell-specific drug-delivery vehicles for cancer therapy served as a template for the design of our entire project.
Left- Dr. Vipul Gujrati
Right- Screenshot of the meeting with Dr. Vipul Gujrati
Dr. Ferreira is a junior researcher at the University of Minho. She is one of the authors of "Selection of a new peptide homing SK-BR-3 breast cancer cells". In this paper, they discovered a peptide, Pep-1, which selectively binds to the HER2+ breast cancer cell line SK-BR-3. Our meeting provided us with insights about the functioning of the peptide and its potential for us in a targeted delivery system like Duonco.
For successful delivery of the THP-OMV to the cancer cell, binding of the THP is not sufficient. Subsequent internalisation is also necessary. No experiments were done at physiological temperatures with Pep-1; hence its behaviour with respect to internalisation was unknown. She suggested that since other THPs generally internalise the following binding, it is likely that Pep-1 will as well. We discussed the difficulties associated with culturing the cell lines mentioned in the paper (MCF-10-2A and MCF-10A) and explored alternative cell lines that could be used to test Pep-1. From our discussion, HEK 293 emerged as an appealing alternative, given that it does not usually overexpress the target markers of Pep-1. Pep-1 contains cysteine at the C and N terminus. Dr. Ferreria helped us understand the role of these residues in the binding ability of the peptide by conferring a loop structure to it. She also mentioned that she has been utilising homing peptides for vesicle-based delivery and that the requirement of a disulphide bond should not be an issue during protein expression in a bacterial system [5].
Dr. Debora Ferreira
Ms Monica P. is a PhD student at IISER Thiruvananthapuram, working under the supervision of Dr. Natesh Ramanathan, Assistant Professor, IISER Thiruvanathapuram. We intend to engineer two types of vesicles, Affi-OMV and THP-OMVs. Affi-OMV focuses on HER2, a commonly targeted receptor, whereas THP-OMV focuses on CX3CR1. Given the time constraints, she suggested that the team prioritise the production of the THP OMVs and characterise them as they were a novel component of our project with minimal background in literature. She also provided us with numerous practical suggestions regarding the planning and execution of our wet lab work. We followed many of her suggestions in order to maximise our efficiency in the lab.
Ms. Monica P
Dr. Chamakuri is an Assistant Professor in the School of Mathematics at IISER Thiruvananthapuram. He suggested trying more differential equations in the mathematical modelling part of the project. He pointed out the practical difficulties of our work plan and said that forming new equations can be challenging for the short period of time the project. He suggested reading more literature. We made changes according to his advice, and we dug deeper into the literature to have a clear idea which later proved to be useful.
Left-Dr. Nagaiah Chamakuri
Right- Screenshot of the meeting with Dr. Nagaiah Chamakuri
We had several doubts regarding various aspects of mathematical modelling, including the suitable model for each system.
The Frontier Symposia in Biology are annual, 2-day conferences organised by the School of Biology, IISER Thiruvananthapuram. This year the symposium was held from 29th April to 1st May, 2022 at our campus. The meetings were a platform for discussing cutting-edge research topics in all fields of biology. Students and researchers from across the country came together to meet and discuss science.
Our team was fortunate enough to attend the conference and meet with Dr. Manidipa Banerjee, Prof. Dipshikha Chakravorty, Prof. Gaiti Hasan, Dr. Bushra Ateeq, Dr. Shantala Hari Dass, and Dr. Devendra Singh to discuss our project proposal. We were able to evaluate our options and study alternative drug delivery carriers thanks to our interactions with them before deciding to finalise OMVs as our drug carrier. After this encounter, our team had a project presentation with Prof. Dipshikha, and she accepted to be our external mentor.
Slide 1. Group photo at the symposium Slide 2. Interactions with the speakers
Summary
All of our interactions with experts aided us in selecting each element of our project. We also got a direction regarding the kind of cell lines we can work on. Through these interactions, our project became more precise and became ready to be taken to its next stage of building it.
With the need for our solutions validated and project design in progress, we started looking at the detailed work plan of the project. We required assistance in selecting the production procedure for the isolation of OMVs, choosing the epitope tags and characterising the OMVs, choosing the appropriate media and controls for our cell line work as well as guidelines for the sponsorship team. Thus, we decided to reach out to experts in these niches and seek their opinion.
Dr. Denice Bay is an Assistant Professor in the Department of Medical Microbiology & Infectious Diseases at the University of Manitoba. She worked with OMVs to understand their formation associated with cationic antimicrobials and their application to reduce antimicrobial resistance (AMR) infections. Interaction with Dr. Denice helped us to optimise our OMV isolation protocol, as her previous work included isolating OMVs from bacteria. She advised us to follow centrifugation rather than ultrafiltration as filters are expensive. From her experience isolating OMV, she was confident that culturing with antibiotics will not result in impacting the overall yield of the OMVs, and proposed that we store the OMV pellets at -20 degree celsius. She also recommended the use of OmpA Antibody for OMV detection which we then implemented to be our preliminary confirmatory step after isolating the OMVs. Throughout our OMV isolation, her counsel was crucial.
Left- Dr. Denice Bay
Right- Screenshot of meeting with D. Denice Bay
We approached Dr. Kurapati to clarify our queries on drug delivery. Dr. Rajendra Kurapati is an Assistant Professor in the School of Chemistry at IISER Thiruvananthapuram, Kerala, with previous experience in nanoparticles and drug delivery. To confirm the isolation of OMVs, he suggested performing DLS followed by TEM. He introduced the possibility of other receptors, such as Folate receptor alpha, being present in epithelial cells and overexpressed in cancer cells. This helped us identify the other potential receptors that can be targeted with OMVs in the future, given that folate receptors are overexpressed in triple-negative breast cancer cells. Read More
Dr. Rajendra Kurapati
Prof. M.R.N. Murthy is a renowned crystallographer who retired as a professor of molecular biophysics from the Indian Institute of Science, Bangalore. He is currently the Distinguished Professor and AstraZeneca Chair at the Institute of Bioinformatics and Applied Biotechnology. Prof. Murty raised valid concerns about the formation of OMV, its solubility, its contents, the immune system’s response mechanism to it, and the structural similarities of its HER2 target. He also suggested performing Cryo TEM to visualise OMVs better because heat from TEM can lead to membrane structure deformation. Since we met him during the inception of our project, we were able to reflect on his questions and design the implementation of our project accordingly.
Prof. M.R.N. Murty
We met Prof. Chakravortty to perform Cryo TEM as the facility was unavailable in our institute. She guided us to Dr Somnath in IISc. Prof. Chakravortty, a renowned microbiologist and pathologist, works as a professor at the Indian Institute of Science (IISc), Bangalore. Through the course of the project, she has offered us several valuable suggestions and acted as our external mentor.
She suggested not using the electroporation method to insert prodrug and enzyme, as it will be difficult for large molecules to get internalised. She also suggested loading our THP-OMV with the mRNA coding for the enzyme instead of the enzyme itself. This aided us in investigating other approaches for effective and rapid internalisation.We decided to read more about this and choose an efficient system. She suggested an easier alternative to verify the internalisation of the OMVs that could serve as a model for the prodrug-enzyme system using small dye molecules as cargo to monitor internalisation. Due to time constraints we were unable to perform this experiment, however we intend to employ this in the future in order to demonstrate the internalisation of OMVs.
Left-Prof. Dipshikha Chakravortty
Right- Meeting with Prof. Dipshikha Chakravortty at IISc Bangalore
Dr Somnath Dutta is the Assistant Professor at MBU at the Indian Institute of Science (IISc), Bangalore. According to Prof. Chakravotty’s guidance, we met with Dr Dutta. He helped us perform Cryo TEM on our isolated OMV samples This was a crucial confirmatory step for our project, that allowed us to proceed forward with further characterisation steps.
Dr. Somnath Dutta
Dr Santhosh working at Rajiv Gandhi Centre for Biotechnology, is a molecular biologist whose primary area of research is to understand the molecular mechanism of chemoresistance and tumour recurrence in triple-negative breast cancer cells. He highlighted the possibility of using different epitope tags like V5 and 3XFLAG for confirmation for the expression of our fusion protein on our OMVs. He brought to our attention that the myc tag, part of our fusion protein that is expressed on our OMVs and the endogenous c myc protein that is known to be expressed in SKBR-3 ( test cell line) have several regions of sequence homology. He encouraged us to verify if the components of SKBR-3 cross react with the anti-myc antibody, and if so, to change the tag used in our construct from myc to V5.
Left- Dr. Santhosh Kumar
Right- Screenshot of meeting with Dr. Santhosh Kumar
We intended to demonstrate the internalisation of OMVs and payload release using mammalian cell lines. We had several questions associated with choosing the appropriate test and control cell lines, culturing conditions and treatment conditions. Hence, we interacted with a number of professors working with breast cancer cell lines. We met with Dr Ramray Bhat, Associate Professor at the Indian Institute of Science (IISc), Bangalore. He guided us on the proper usage, viability and availability of different cell lines. In his opinion, HMLE cell lines are easier to maintain in culture as compared to MCF-10 A. Read more.
To understand more about MCF-10A cell lines, we contacted Dr Medhavi Vishwakarma. She directed us to doctoral candidate Ms Sindhu M. She highlighted the difficulties while working with MCF-10 A. The medium for culturing required several special components, reaching confluent levels takes quite long. Due to these difficulties associated with MCF-10A we decided to work with an alternative control cell line (HEK 293). The professors also reaffirmed our decision to work with SK-BR-3 as our test cell line. We then interacted with Dr Rakesh S. Laishram. He gave inputs on the quantification of OMVs and showed the possibility of using fluorophores for the detection of internalisation of OMVs.
Prof. Suresh Das is an emeritus professor at IISER Thiruvananthapuram. He was formerly the Executive Vice President of the Kerala State Council for Science, Technology and Environment.
We had planned outreach and communication initiatives. However, it was challenging to acquire the necessary funding for these endeavours. Prof. Suresh Das advised us on how to secure the financing of our proposed study. His suggestion to prepare a detailed budget breakdown assisted us in prioritising substances. We created a sample budget, which aided in the launch of our project.
Summary
By consistently meeting with experts, troubleshooting with them and going through literature to resolve their concerns and questions, we were able to define a detailed plan for the execution of our project. Integrating their insights helped us greatly improve the scope of our project.
Not only did we want to connect with professionals in this field, but we also wanted to reach out to other people and explain our initiative. We desired their opinion to understand the social landscape on the premise of breast cancer and drug delivery. Knowing their expectations and level of awareness felt necessary to streamline our outreach activities and project design. From our conversations with stakeholders and analysing our survey results and interviews, we realised that people weren't aware of the diagnostic facilities available to them, when to get tested, or the available treatment options. We also discovered that financial constraints prevent people from getting tested in economically backward households. A general lack of proper awareness was found among people, along with the several taboos and stigma surrounding breast cancer, e.g., Men can't get breast cancer.
Our team thought it would be a great idea to have an unveiling of our project, to introduce it to the community of IISER TVM. So on World Cancer Day (February 4, 2022), we used a fun interactive mystery questionnaire to disclose the project's theme, and students could guess it by filling out a Google Form. To promote our initiative, we also placed posters throughout the campus and had a selfie competition with the poster.
After much anticipation, on May 16, 2022, we formally introduced our project concept and team to our institute's faculty, staff, and students. Read More.
Each team member introduced the different aspects of the project, the structuring of the subteams, and our detailed work plan for the next few months. We also had an interactive session to address their questions and discuss the points in more detail. We wanted this event to promote the iGEM culture in our institute, and encourage students to form future iGEM teams. During the event, we also unveiled OMIVON, our team mascot.
With the help of the Mentimeter website, which incorporates user ideas, we got to learn which social media platforms the audience prefers and how they view synthetic biology. Later, we planned and designed our human practices and outreach initiatives using these inputs. The previous iGEM team from our institute also had a discussion about how iGEM had aided their development a year earlier. Additionally, we got a lot of feedback from the audience, which was helpful for the project.
Slide 1.Presentation during Project reveal
Slide 2.Faculty and students attending the presentation
Slide 3.Team
Slide 4.Team explaining synthetic biology during presentation
Slide 5.Audience view on Synthetic biology
We spoke with the housekeeping staff of our institute. We interacted with them to know their level of awareness of breast cancer. To further see if they have undergone any detection tests earlier. Most were aware of cancer detection tests like mammograms, but none had undergone them. They were also aware that breast cancer might be associated with a lump. But they were not sure how to detect them correctly. We arranged an awareness talk for them, and they were eager to attend. We also facilitated a preliminary screening for breast cancer using iBreast for some of the staff who attended the talk.
Team interacting with housekeeping staff
Dr. Hema Francis is the medical officer at IISER Thiruvananthapuram and the secretary of IMA (Indian Medical Association), Nedumangad. We conversed with her about the lack of knowledge and the rising incidence of breast cancer in our region. She has organised numerous screening camps in rural areas and proposed that we conduct a similar campaign on our campus. She helped in organising a breast cancer screening camp in our institute. She also assisted us in contacting individuals for other events, such as teaching students at nearby schools and getting medical professionals.
Based on our interaction with the housekeeping staff, we organised a successful breast cancer screening campaign for our institution's housekeeping staff. IMA (Indian Medical Association) and KIMS Health Centre, Thiruvananthapuram, collaborated with us on the event. Dr Hemalatha assisted in the organisation of this programme. Dr Kavitha Ravi, the National Secretary of the Women Doctors Wing in IMA, was the guest speaker during the event. She is an adjunct professor at Government Medical College, Thiruvananthapuram. The examination was administered utilising the iBreast screening device. iBreast is a portable gadget that detects breast abnormalities. It is not a confirmatory test, although it can detect abnormalities. Read More.
Slide 1. Inauguration ceremony for breast cancer screening camp Slide 2. Team along with technicians and other medical professionals Slide 3. Participants filling consent form before undergoing screening test
Through activities such as surveys and interviews with the public, we determined that the public is unaware of breast cancer and how early detection and good knowledge can save thousands of lives. Lack of awareness prevented individuals from getting screened for breast cancer, resulting in late-stage diagnosis. Compared to early detection, the percentage of cancer patients who are cured is substantially lower when the disease is identified at a later stage. Consequently, we saw the need for awareness. We organised a campus-wide breast cancer awareness lecture for our institution's housekeeping staff. Dr. Hema Francis, medical officer of IISER Thiruvananthapuram, delivered the lecture. So that the audience could comprehend, it was conducted in the native tongue (Malayalam). The discussion included breast cancer examination procedures. Read more.
Slide 1. Dr. Hema Francis giving an awareness talk on Self Breast Examination Slide 2. Housekeeping staff attending the awareness talk
iBreast is a preliminary screening test for breast Cancer. A few members of our team spoke with an iBreast technician to learn how people are scanned at camps, whether people are reluctant to be screened, what challenges they encounter during the procedure, and what limitations the technique has. Read more
For confirmation, a mammogram or FNAC is required. The main aim of organising screenings is to reach middle-class household women who don't prioritise their health over their family and chores. The system involves a portable sensor and a Bluetooth-connected screen to view the image of 1/16th of the area at a stretch. Gradually we scan the entire area in parts and complete the scan. If there are any lesions or lumps, it will show a red colour as an indication. There are a few situations where false positive cases are shown. However, this method has proven quite helpful for screening a number of patients and assisting them in receiving an early diagnosis of their ailment.
Overall, she claimed that despite people's initial reluctance, after learning about cancer and the need for screening, they are prepared to undergo screening and be informed of developments by the doctors. This was an inspiration for us to conduct more activities in the future [6].
Slide 1.Ms Reena
Slide 2.iBreast screening equipment
Our institute is nestled in the foothills of the western ghats, 40 kilometres from the main city of Thiruvananthapuram. As demonstrated by our survey, interaction with the staff of our institute and other stakeholders there is a significant lack of awareness among our local community. People in the vicinity of our institute reside in rural areas and lack access to screening tools.
Therefore, we decided to educate them on self-breast inspection techniques that can be undertaken in their homes. We distributed pamphlets on campus, in the neighbourhood near campus, and in our hometown. Brochures were available in English and three regional languages (Malayalam, Tamil, and Hindi). Illustrations were utilised to emphasise the SBE (Self Breast Examination) approach. It was created so that people may easily comprehend and follow the instructions. It was verified by Dr. Hema Francis before distribution. Read more
Feedback: Distributing pamphlets was quite useful as everyone could not attend the awareness talk that we conducted in our institute. The staff of our institute even collected pamphlets for their relatives. Distribution of these in public places was also effective.
Pamphlets in Tamil, Hindi and Malayalam are available here.
We desired to expand the scope of our awareness campaigns. In addition to expanding the programme, our objective was to spread our message to people of all ages and genders. We decided to stage a play in public. We, in collaboration with the Theatrics society of IISER Thiruvananthapuram, performed a play, "Let's fight with Courage", to the crowd in one of the biggest malls of Thiruvananthapuram city - Mall of Travancore.
Team members and drama club members collaborated to create a play about the significance of early breast cancer screening. Read more
In the drama, a housewife discovers that the lump on her breast may be breast cancer after watching television awareness programmes. Her examination results indicate that she has breast cancer. A few months of treatment were enough to eradicate cancer. It disseminates the idea that cancer screening tests are available and can save lives when used appropriately. We also described how to diagnose breast cancer through our play.
Feedback: The audience considered our performance to be beneficial and appreciated our efforts. In addition, we presented them with pamphlets that explained breast cancer detection in depth.
Slide 1. Team performing drama at Mall of Travancore Slide 2. Drama performance Slide 3. Explaining steps of Self-breast examination during drama Slide 4. Team along with Theatrics club of IISER Thiruvananthapuram
Premi Mathew is the founder and CEO of Hair for Hope India and Protect your mom. She has survived breast cancer. She initiated the Protect Your Mom (PYM) campaign to emphasise the significance of breast cancer screening at an early stage and aims to convey this message to children through whom their mothers will be informed. The mission of Hair for Hope India is to donate hair to cancer patients who have lost their hair as a result of chemotherapy.
Through our interactions with breast cancer survivors, we learned that although hair loss does not cause discomfort, it can significantly impact a patient's self-esteem. Additionally, it influences how others treat them. It is widely understood that one of the side effects of chemotherapy is hair loss. Thus, we visited Premi Mathew and learned more about her business and her fight against cancer. She has encountered several obstacles during her treatment and was aware of the significance of early detection. To support her wonderful cause, we collaborated with Hair for Hope India and the Protect your Mom campaign. We distributed posters conveying the PYM message.
Our meeting with Premi Mathew prompted us to organise a campus-wide hair donation drive. Cancer therapy takes a toll not only on the physical health of the patient, but also on the mental health. Patients often lose self confidence in their physical appearance due to mastectomy and hair loss due to chemotherapy. Using wigs to restore their confidence might be of tremendous assistance. Approximately twenty ponytails of hair would be needed to create a wig. On August 15, we held a hair donation event with the non-profit Hair for Hope, India. Read more.
We sent information regarding the hair donation drive to the students and faculty at our institution. During the campaign, several students donated their hair. We obtained a written MoU (Memorandum of Understanding) from the CEO of the non-profit organisation to prevent any misuse of the donated hair. We ensured to receive the consent from the participating individuals. We presented the donors with bookmarks as a token of our appreciation. We also informed them about the PYM (Protect Your Mom) campaign and how early breast cancer screening can save lives. We packaged the hair and shipped the package to the Amala medical college in Thrissur, where the donated hair will be transformed into wigs for people with cancer.
Hair donation campaign in collaboration with Hair for Hope
Dr. Shantala is a neuroscientist and science administrator who is an Associate Director at IndiaBioscience. She is a passionate researcher who is also interested in scientific communication. We had questions regarding how to advance our project's research and communication components. She provided strategies for communicating synbio to non-STEM audiences. She encouraged students to take classes in synthetic biology, as they are the next generation and synthetic biology is a booming subject. We had a list of events planned for the outreach portion of our mission. She instructed us to prioritise the essentials and implement them after appropriate planning. After speaking with her, we devised a systematic plan for the outreach initiatives. We decided to offer synbio lessons to school children. Her advice to focus on school students proved helpful, as the questions posed by students throughout our interactions with them expanded our creativity even more.
Left- Dr. Shantala Hari Dass
Right- Screenshot of the meeting with Dr. Shantala Hari Dass
Some of our stakeholders were kind enough to participate in the women's talk series 'We-Talk', which we, along with other iGEM Teams from India, organised. Talks highlighted various issues faced by women. Speakers of our talk series include:
The topics covered were Menstrual Hygiene, Adolescent Challenges, Cervical Cancer, and Breast Cancer. Their presentations on these topics enlightened the audience and proved helpful. After the three-day lecture series, a panel discussion took place. Each team's collaborators participated and discussed the issues women face. Students from different colleges attended the talk. The interactive sessions at the end of each session were very beneficial.
As the lab work for our project progressed, we felt it essential to acknowledge the biological waste we generated and ensure the proper and secure disposal of the biological wastes produced in our laboratories. The fact that our campus is located in a biosphere reserve in the Western Ghats makes it incredibly important that we pay attention to how we handle the disposal of biological waste. We conducted in-depth research and a number of interviews with individuals involved in the biological waste management system. Read more.
It all started with a tip to the red coloured bin, leading to the question of why there are different coloured bins and a different disposal method for each one.
We talked with: Dr Poonam Thakur - Lab Safety officer; Mrs Nithya Rani NM - Teaching Assistant; Miss Anjitha and Miss Akhila - Housekeeping Staff; Jishnu - IMAGE Vehicle Driver; Mr Peter Sammuel, Manager in operation; Naveen M - MEP Engineer(Mechanical, Engineering and Plumbing Engineer, Dr Joseph Benaven, the former secretary of IMAGE, Dr V.Vijayachandran sir, secretary during the early stages of IMAGE plant.
The details of our interaction and our findings throughout the course of our investigation into the waste produced by our institute is attached here.
The Indian Institute of Science in Bangalore hosted AIIM, the national gathering of all Indian iGEM teams, during July 23–25, 2022. AIIM gave us a forum where we could talk, work together, and improve our presentation abilities while also picking up new information from subject-matter specialists. All the teams got a feel for the final project presentations thanks to the mock jamboree-style presentations held for each group.
The AIIM turned out to be incredibly constructive for us. We also had the opportunity to respond to, incorporate, and develop the judges' queries and ideas into our product. Our team had a wonderful experience participating in the AIIM.
Slide 1. Team at IISc Bangalore Slide 2. Poster presentation Slide 3. Team after presentaion Slide 4. Presentation at AIIM Slide 5. Group photo of all teams participating in AIIM
Summary
Thus, we were able to establish an empathetic dialogue with our stakeholders. We contributed to the cancer survivors with a hair donation campaign; we helped the housekeeping staff at our institute get access to a preliminary screening; we were able to educate the general public about breast cancer through our drama, pamphlet, project reveal and other events. In short, we took positive strides in bridging the gap between science and society.
Safety has been an integral part of our project from the very beginning. To ensure that we did our part in being responsible researchers, we interacted with a few people and also did a thorough literature review to learn about the safety rules and laws governing the usage of genetically modified organisms. These interactions helped us prepare consent forms and acquire safety approvals wherever applicable.
Dr. Poonam is an Assistant Professor at IISER Thiruvananthapuram. She discussed with us the protocols of waste disposal and also guided us throughout all our safety aspects during the project. She was instrumental in our ‘Track Your Waste’ initiative and also helped us review our project’s safety form and protocols. As the biosafety officer for our institute, Dr. Poonam also conducted a biosafety workshop on safety and security where she trained us adequately to be wary of the safety aspect of our project. Read more.
The topics covered were:
The seminar was truly helpful and helped us prepare and plan our wetlab in the safest possible way.
She also prompted us to meticulously research on the present legislations and regulations set by the Ministry of Science and Technology's Department of Biotechnology (DBT).
The DBT supports and oversees biotechnology development, as well as establishes guidelines for safe practices and laws for managing genetic engineering in India. Their Biosafety Research Programme emphasises the implementation of biosafety procedures, rules, and guidelines under the Environment (Protection) Act 1986 and Rules 1989 to ensure user and environmental safety from the use of Genetically Modified Organisms (GMOs) and products thereof in research and application. The programme mandated a two-tier process consisting of Institutional Biosafety Committees (IBSC) at the Institute/company and the Review Committee on Genetic Manipulation (RCGM) in the Department of Biotechnology. As a result, we were able to obtain the necessary instructions from their online resources and apply for IBSC and RCGM approval.
Dr. Poonam Thakur
Dr. Harilal Madhavan is an Ad-Hoc faculty at IISER Thiruvananthapuram, Kerala. He is a development health economist specialising in economic and interdisciplinary research on Asian indigenous medicines, health and pharmaceutical policies and intellectual property rights.
During our meeting, we discussed our plans about releasing a survey to analyze public awareness, conduct camps and other activities we had in mind. He also introduced the topic human subject research to us and told us the importance of complying by it during all our events and since then we ensured to follow necessary guidelines to respect the interest of our participants. Read more.
Our team held a number of online and offline meetings, webinars, polls, interviews, films, camps, and competitions throughout the course of our project to raise awareness and gather information for improving it. We made sure to take all necessary steps and adhere to all applicable laws and regulations, as well as any institutional guidelines or instructions, that concerns Human Subject Research.
Dr. Harilal Madhavan
He is an Associate Professor in the Chemical Engineering department at the Indian Institute of Science (IISc) and also an Associate Faculty at the Centre for BioSystems Science and Engineering.
We met with Dr. Rahul to gain insights into how to safely engineer cytolysin A as a display system for our OMVs. In the past few years, he has worked intensively with ClyA to determine how natural ClyA can operate as a toxin against mammalian cells. He provided the following recommendations:
This mutation helped us secure the required level of safety for our Cytolysin A study.
Left: Dr. Rahul Roy, Right: Meeting with Dr Rahul Roy
Summary
Through our efforts, we were able to scrupulously follow the general safety requirements, safety guidelines prescribed by our institute, iGEM’s safety rules and policies throughout the execution of our project. We were able to develop a solution that is safe both for the environment and the people who depend on it.
The allotted eight months of the iGEM competition is limited time to explore the project's full potential and explore the wide ranging applications of Duonco. Further several stages of clinical trials are necessary before Duonco can be used as a fully functional drug delivery system. We met with experts from various fields to learn more about how the scope of our delivery system can be extended, and the experiments and trials that are required to do the same.
Dr. Rona is an Assistant Professor at the department of oncology at RCC (Regional Cancer Centre). We discussed the adverse effects of chemotherapeutic and immunotherapeutic treatments during our meeting. She urged us to continue developing our project and informed us of the other available targeted therapies that target antibodies. T-DM1 is an FDA-approved, effective drug for treating HER2-positive breast cancer. In addition, GMO-based pharmaceuticals are also available in the market; however, none of them is ideal. She stated that one of the side effects of antibody-directed therapy is the development of resistance to the drug. Furthermore, to determine whether our treatment has similar side effects, it must undergo clinical trials during which most drugs are typically rejected. Only after multiple stages of testing can the efficacy of a particular treatment be confirmed.
Dr. Rona Joseph
Dr. Priya is a scientist at the Rajiv Gandhi Centre for Biotechnology (RGCB), Thiruvananthapuram. She is working on understanding the mechanisms leading to the early onset of breast cancer. She was able to answer several questions we faced while taking our project further.
Dr. Priya Srinivas
Dr. Ram Mohan is an established scientist working in the field of nanomedicine to aid in diagnosing and treating diseases. He works as a scientist at Rajiv Gandhi Centre for Biotechnology (RGCB), Thiruvananthapuram. He suggested substituting exosomes for OMVs as a medication carrier. In addition, he proposed chemical conjugation for loading peptides onto the surface. Electroporation, sonication, and CaCl2 are the several methods he proposed for loading drugs into vesicles. The stability of the vesicles is compromised by electroporation, per his recommendation. He is familiar with employing CaCl2 for internalisation. When we inquired about his opinion regarding the mode of administration, he stated that it depends on the optimisation technique. He emphasised the significance of half-life and instructed us to determine the half-lives of the prodrugs and enzymes we would be working with. As we are working on the preliminary stages of the project, we considered his recommendations and adopted those we deemed feasible for our level.
Dr. Ram Mohan Ram Kumar
Ms Varsha Jaisimha is a former IISER Pune iGEM team member and also participated in After iGEM for a year. She is currently a member of the iGEM Foundation's engineering committee. She is accustomed to presenting iGEM work to a variety of audiences. We sought her since we were uncertain of how to present our project on a big stage. She provided recommendations for the poster and the presentation. Even while we included human practices in other aspects of our project, such as the wetlab and dry lab, this may not have been adequately communicated during the presentation. During presentations, she urged us to make a special effort to highlight our collaborative efforts. Presentations require effective time management. Within the presentation time provided, the project must be presented effectively. During our presentation, we will be sure to address these concerns. We later attended Ms Varsha's presentation at the All India iGEM Meet.
Ms Varsha Jaisimha
She is the Director of Market Access at Sanofi India Ltd and was previously the Director of Oncology and Transplantation at BU Head Oncology and Transplantation, Sanofi India Ltd.
Ms Sangita has 20 years of career experience in the oncology department in the Indian market. She was part of the pioneer team that introduced high-dose chemotherapy in India as a need in cancer therapy. Later, she joined Sanofi Aventis, a French multinational pharmaceutical and healthcare company. It engages in the research and development, manufacturing and marketing of pharmaceutical drugs, principally in the prescription market) and is currently working in the Market access department. Read more.
Interaction with ma'am was an eye-opener for us to the marketing world and its workings. She gradually took us through all the phases a product had to undergo before reaching people. She briefed us about the significant steps involved.
We had questions about how to proceed with our project in the future, and we learned that the cost we fix would vary depending on whether the project is in its early phases and whether it requires a licence or a proof of concept. We didn't see how a drug delivery system would be different. She enumerated the pros and cons. Bioequivalence tests are needed. Using an existing medicine increases safety, but we must first pass an inferiority trial. But this would reduce failures in phases one, two, and three trials. She also discussed how a delivery system could obtain an IP patent if the conceptualisation can be proven. She advised us to refer to the CDSCO, Drugs and cosmetics act under DCGI. The act requires clinical trial phase three in the intended country or a waiver promising phase four. We were also introduced to a commercially available nanoparticle-based targeted delivery method, Abraxane, to refer more and develop our future direction.
Ms Sangita Topiwala
Kerala Startup Mission is an agency started by the Government of Kerala to support rising entrepreneurs in Kerala. At the Kerala Technopark, we had the opportunity to discuss our concept with Mr Anoop Ambika, CEO of Kerala Startup Mission. He was interested in our proposal and advised that we discuss the process of forming a company, determining who would be the CEO and dividing the stakes among the team members. He also suggested that we approach the Startup Mission via their "Idea Grant" for student startup enterprises to receive the initial funding required to finish our proof of concept work and give us a push to begin licensing our project to interested companies or programs like that of C-CAMP's DIA mentioned earlier.
At Technopark, Thiruvananthapuram
During the All India iGEM Meet, we had a round table chat where we could pitch and discuss our proposals with various founders of Indian biotech firms. These individuals included CEOs, corporate executives, and senior entrepreneurs. The teams interacted for around 15 minutes at each round table, establishing interesting talks about turning ideas into commercial businesses. We had a comprehensive chat with Dr. Vaibhav Bhatia, founder of Lamark Biotech, who offered us insights into the process of patenting and licensing your ideas as researchers moving into the industry.
Slide 1.Round table conference with Dr. Vaibhav Bhatia Slide 2.Pitch in of our project
Summary
By building on these discussions we could design our future experimental plan. They also emphasised the importance of carrying out preclinical and clinical studies and how they are crucial for determining the safety and efficacy of our system. We also obtained insights on building a road map for entrepreneurship and market entry of Duonco.
Each step in the project was the product of extensive literature review and numerous conversations among ourselves, experts, and stakeholders. Human practices are inevitable in any iGEM project since they bridge the gap between developers and society. Our different interactions helped us discover the several layers and complexities associated with the problem we are trying to solve.
We realised that as student researchers we often lose sight of how our product could impact our stakeholders. Interacting with them provided us with the grounding necessary for our research work, and made us aware of the broader implications of our project. Our initiative is still in its early stages, and we hope to meet various experts and stakeholders we have not reached out to in the future phases.
