Inclusivity

IRIS is inclusive. Nowadays, the cost of cancer treatment often outreaches the average monthly salary. For example, it is estimated that the average total cost of a 6-month course of chemotherapy is almost 27,000$³.
Unfortunately, these costs are impossible to be paid by some people. Cancer death rates are approximately 20% higher among residents of the poorest U.S. countries compared to the most affluent countries.²

This question led us to further research about cancer. In 2023, 1,958,310 new cancer cases and 609,820 cancer deaths are projected to occur only in the United States.¹ This, combined with the global economic crisis, made it clear that the finding of a new, effective and affordable cancer treatment was crucial. We knew that, in 2023, it’s finally the time to put an end to this problem that causes so much pain. And, just like this, we had ended up with a project focused on cancer treatment. Every scientist in the world wishes to find a way to cure this global, fatal disease. Our team believes that synthetic biology is the key to make this wish come true, and this was the main thing we had in our minds when designing our project.

Furthermore, the cost varies depending on cancer type. The estimation of prostate cancer treatment is about 8,181$ - 8,586$, while leukemia treatment can reach up to 16,551$. ³ IRIS is a therapeutic tool for all types of solid tumors, and the way that is administered is somewhat the same for every case of cancer. Our bacteria and a luciferin analogue, methoxy e-coelenterazine, are administered as close to the tumor as possible, and they find their way to the cancer cells. Just because IRIS consists only of our bacteria and the luciferin analogue, it is an easily administered and affordable therapeutic approach. That makes it highly accessible, even to areas that lack high-end medical infrastructure and developed health care systems.

Safety

Our priority as a team was to design IRIS as a both effective and safe project. The final step of IRIS’ mechanism of action is ROS production within cancer cells. ROSs cause are cytotoxic, damaging cell proteins, DNA, membranes and organelles, destroying tumors completely. The good news is that healthy cells have their own mechanisms to beat ROSs off, and they aren’t as much affected as in other kinds of treatments.

But what other factors propel our project closer to becoming a safe therapeutic approach?

The utilization of E.coli, bacteria which are already part of the human microbiome. Although bacteria-mediated tumor therapy is being tested and seems to be giving promising results, directly injecting bacteria in the human body can indeed trigger side-effects. It can lead, for example, to inflammation or other forms of immune responses. That’s why our team proposes the employment of a Drug Delivery System (DDM), a liposome for example, to deliver and release IRIS to its target. Other safety parameters we have thought that would help IRIS reach the goal of being a proper therapeutic tool are: the integration of Adhesive Proteins to improve IRIS’ targeting ability, the employment of a Quorum Sensing System that controls the starting point of IRIS’ genes expression within the tumor microenvironment and the design of a Kill Switch Mechanism that kills our bacteria after the treatment is done.

Our goal is to make IRIS as safe as possible, so that people trust it as a future therapeutic tool.

Our Vision

iGEM team IOANNINA strives upon building and introducing a safe, accessible, cost-effective and less equipment-dependent treatment option for solid tumors. We strongly believe that our project has the potential to positively impact the lives of countless individuals facing cancer, regardless of their geographic location or available resources.

Step 1: Where it all began

iGEM Ioannina 2021

iGEM Ioannina 2021 x iGEM Ioannina 2023

We held several meetings with members from the team iGEM IOANNINA 2021. Most of these members have now graduated and embarked on master's degrees and phDs.
They supported our team in its initial stages and provided guidance during our brainstorming sessions. Their questions were categorized into specific axes:

-What are the current problems in our society?

-Which one does your team find intriguing to solve?

-How can Synthetic Biology help solve this problem?

When we presented our initial idea to them, they had a lot of questions to ask regarding its in vivo applications. These questions motivated us to search a little further on finding solutions to possible problems:

-How can IRIS be administered as a therapeutic product to a patient?

-Is IRIS safe enough for healthy cells? How can we increase its safety?

-How can our approach be more specific to tumors?

They also helped us select an assembly method, by analyzing the advantages and disadvantages of HiFi DNA assembly and Golden Gate assembly. Let's not forget their contribution in fundraising, as they gave us some precious advice.

Photodynamic Therapy

While we were still brainstorming about possible project ideas, a meeting with professors from the University of Ioannina took place, as they wanted to learn more about our thoughts and help us with making decisions. Specifically, the professors that attended the meeting were H. Stamatis, Biotechnology professor and one of our Primary P.I.s Mr K. Papaloukas, Bioinformatics professor at the Department of Biological Applications and Technology Mr P. Doulias, Biochemistry professor at the Department of Chemistry Mr A. Tzakos, Organic Chemistry professor at the Department of Chemistry.

Until that meeting, our thoughts were focused on developing a therapeutic approach for acne. This approach was somehow based on porphyrins, something that reminded mr A. Tzakos about Protoporphyrin IX as a PhotoSensitizer molecule. This idea led us to study in depth about PpIX and its applications in PDT, marking the first step in redirecting our focus from acne to the treatment of cancer by using PDT.

Step 2: Lab advice

Tarvi Teder- Low copy-number plasmids and Drug Delivery Systems

Tarvi Teder, a PostDoctoral researcher in Biophysics at Karolinska Institutet, provided us with precious advice on how to isolate low-copy plasmids, as we encountered issues while isolating pBBR1-MCS2. After explaining our problem, he was confident that our failed attempts of isolating the plasmid had something to do with the plasmid’s antibiotic resistance and advised us on checking if we indeed had the right plasmid to start with. His advice was precious and led us to start a troubleshooting procedure for the isolation of pBBR1-MCS2.

You can find the troubleshooting procedure HERE

He also found the direct injection of active bacteria in the human body risky as directly injecting bacteria in a person’s body can trigger endogenous immune responses. The bacteria are likely to be recognized as threats, and that could lead to immune activation (leukocytes, such as neutrophils and macrophages are recruited to the site of infection to destroy the bacteria) or an inflammatory response (swelling, redness, heat and pain) at the injection site.

A possible solution to that issue would be the utilization of a drug delivery system. Liposomes, for example, can be used to deliver IRIS directly to the target tumor site, where a triggered release of the bacteria could take place. In that way, the bacteria are not going to directly interact with the patient and possible immune responses can be avoided.

Step 3: Iris as a cancer treatment

Barry Campbell- targeted therapy

Barry Cambell, professor in Infection Biology and Microbiomes at University of Liverpool, had a conversation with one of our members about the role of adhesive proteins and how we can incorporate them to our bacteria for a more targeted and safe therapy. Adhesive proteins can bind to glycosylated receptors that exist specifically in cancer cell membranes. This binding would lead to the closer secretion of our bacteria’s factors directly into the cancer cell, and would therefore minimize the possibilities of untargeted secretions.

He also suggested possible methods that can be employed for inserting bacteria into a cancer cell, in order to minimize the loss of light emission and important molecules in the intercellular space and therefore increase the project's effectiveness. His suggestions are valuable for the future development of IRIS.

Department of Medicine × iGEM Ioannina

Mr Kolletas, Mrs Eugenia Roupakia and Mrs Simone with members of iGEM Ioannina wet lab

In September, we had an insightful conversation with Mr Kolletas, Biology professor at the Department of Medicine of University of Ioannina, Mrs Eugenia Roupakia, Doctoral Student in Molecular Cellular Biology at the Department of Medicine of University of Ioannina, and Mrs Simone Besta, PhD candidate in Molecular Cellular Biology at School of Medicine of University of Ioannina.

We presented our project to them and we had a big conversation about the safety of our project. They agreed that an effective way to a more targeted therapy is including adhesive proteins and even suggested several other molecules that we can employ to increase the target specificity of our mechanism.

We also talked about possible kill-switch mechanisms to further increase our project’s safety. Mr. Kolletas, specifically, had many suggestions about this topic, including a kill switch mechanism inducible by our bacteria population, or by an σ³⁸ rpoS factor, which regulates gene expression in E. coli.

Another outcome from that meeting was the highlighting of the importance of knowing methoxy e-coelenterazine’s half-life, as it would better regulate our therapy and its future implementation. The idea of co-culturing our engineered bacteria with breast cancer cells to test the proof of our concept seemed to puzzle Mr. Kolletas, who noted that some subtypes of breast cancer have mechanisms for countering oxidative stress. He proposed that we possibly need to make our therapy more specialized to a certain type of solid tumor cell line to achieve better results.

Mrs Kallergi G. - stages of drug development

Mrs Kallergi, and members of iGEM Ioannina wet lab

In October, we had a meeting with Mrs Kallergi, Assistant Biochemistry Professor at the University of Patras, a researcher at the European Liquid Biopsy Society (ELBS) and a former supervisor of the team iGEM Patras Med 2022. Due to her experience with iGEM and her expertise in the field of liquid biopsy for cancer diagnostics, we were happy to engage in a conversation with her.

Commenting on our idea to co-culture the engineered bacteria with cancer cells, she suggested the possible co-culture of our transformed bacteria with healthy human cell lines in order to test possible side-effects of our project.

She explained to us the whole process of drug approval and she mentioned the significance of a precise and clear step-by-step plan for the possible in vivo application of our project in the real world. A logical workflow of in vivo experiments would start by testing on cancer cell cultures, then move on to animal models and after many stages of approvals our experiments could finally continue on clinical trials on humans.

All these steps should be taken very carefully in order to minimize possible risks and ensure that possible positive effects are indeed our project’s effect, rather than an artifact. Finally, she agreed that our therapy must be very well regulated in order to avoid immune responses. Especially with cancer patients who have already received treatment and are under immunosuppression, the possibility of risks occurring is greater, and for that reason, she noted that we should consider even more parameters that would make our therapeutic approach safe.

Step 4: Mentorship Program

A fourth, special category dedicated to our mentor, as she contributed to all the categories above. During our journey, we were lucky to have an amazing mentor and friend by our side, Sofia Oikonomou.

Sofia Oikonomou

Sofia, currently a master’s student at University of Copenhagen, a graduate from the Department of Biology, University of Patras, a former member of the Enzyme and Synthetic Biotechnology Laboratory at the National Hellenic Research Foundation and a team member of iGEM Athens 2022, has all the necessary knowledge and experience to help us improve our project.