Team:Freiburg/Achievements

Overview Motivation CAR T Cells Tumor Microenvironment AND Gate Outlook Achievements References

Members Attributions Partners Contact

Main Project Modeling Applied Design Proof of Concept BioBricks Basic Part BioBrick Improvement Interlab Study

Safety Cloning HIF1A Knockdown Cell Culture Lab Book

Human Practice Integrated Human Practice Collaborations

Achievements

Most important achievements

Knock Down

We designed several shRNAs for a HIF1A knockdown and successfully generated three stable HIF1A knockdown HEK293T cell lines and one stable HIF1A knockdown Jurkat cell line. Additionally we could show, the shRNA does not affect the transiently reintroduced HIF1A in HEK293T cells.

Kill Switch

As an integral part of our Integrated Human Practice, we achieved an improvement in safety of our CARTEL T cells. We generated stable cell lines expressing herpes simplex virus 1 - thymidine kinase, which allows the specific elimination of modified cells by administration of ganciclovir.

Stable Cell lines

Stable cell lines for promoter characterization, knock-down of HIF1A and kill switch were generated via lentiviral transduction and sorted by FACS.

Requirements for a Gold Medal

Integrated human practice

The goal of CARTEL^TM is to create a safer immune-based therapy. Hearing many concerns about the safety of our project and conducting a survey, we integrated further safety precautions. We successfully tested a construct containing a thymidine kinase which makes T cells sensitive to ganciclovir induced apoptosis. We went over a realistic embedment of our project in the context of acceptance and side effects. Additionally, we evaluated possibilities to optimize the manufacturing process of CAR T cells with the aim to contribute to a therapy, which is affordable and versatile in its use for many cancer patients. Finally, a survey was able to confirm the importance and the need for immunotherapies, which have potential to improve the efficacy and safety of cancer therapies.

Improve a previous part or project

Model your project

Modeling was an important part of our project with which we could determine whenever our understanding of the system has a good substantiation. Combining the hypoxia response element, cAMP response element and CTLA4 promoter in our CARTEL^TM AND gate to boost the specificity of the CAR expression, was challenging. After modeling different possibilities and comparing the results we designed an AND gate were HIF1A is knocked out and reintroduced under the control of either the cAMP response element promoter or the CTLA4 promoter. By this usage of modeling, we aim to improve the safety and specificity of emerging synthetic CAR therapies.

Demonstrate your work

Requirements for a Silver Medal

Validated Part/Validated Contribution

Collaboration

Collaborations were an important part of our project, which helped us to improve our own project but also to help other teams to reach their goals. In two meaningful bidirectional collaborations with the teams from the ETH Zurich and Heidelberg, we were able to fulfill this criteria. To help Heidelberg with their mutagenesis project, we performed mutagenesis experiments with E. coli in order to test the mutagenicity of different plasmids, while Heidelberg helped us with the cloning of a plasmid. In the collaboration with Zurich, we performed a killing assay with Azurin on HEK293T cells in order to show the toxicity of the protein used by Zurich for their cancer treatment. In return, Zurich helped us to create a model which shows the time the CAR is still expressed on T cells in the case they leave the microenvironment.

Human Practice

As our project deals with a major research topic, we wanted to involve the society into our project. In order to get an overall opinion about synthetic biology, immunology and cancer we conducted two surveys and got into dialogue with people on the Science Fair in Freiburg and at the Science Days in Europa Park. Hereby, we were able to hear about the concerns about our project, which we used to improve our project. In addition to that, we talked with experts in the fields of oncology and CAR T cells, to discuss whether our project is useful and how we can improve it to be more safe for potential clinical trials.

Requirements for a Bronze Medal

Register and attend

We registered our team, had an amazing summer and we are coming to the Giant Jamboree!

Deliverables

We meet all the deliverables.

Attribution

We created a page on our wiki with attribution of each aspect of our project which as well contains our gratitude toward all the people and institutions that supported us.

Characterization/Contribution

We participated in the Interlab Study.