Claire Wang

BioE PhD Proposal Presentation

9:00 am, Wednesday May 20, 2025

Location: MoSE G021

Advisor: Dr. John Blazeck (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology)

Committee Members:

Dr. Mark Stycztnski (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology)

Dr. Gabe Kwong (Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University)

Dr. Alexander Vlahos (Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University)

Dr. Sarwish Rafiq (Department of Hematology and Medical Oncology at Emory University School of Medicine)

Adenosine-Inducible Self-limiting Armored CAR-T cells for Breast Cancer Treatment

Chimeric antigen receptor (CAR) T cell therapy faces challenges in solid tumors, where the immunosuppressive tumor microenvironment (TME) is a major factor that limits efficacy of CAR-T cells. Specifically, breast cancer is a type of solid tumor that has been shown dysregulated metabolism leading to buildup of immunosuppressive byproducts such as adenosine (ADO). This high ADO environment suppresses the cancer killing function of CAR-T therapy. To address the immunosuppressive environment in TME, recent clinical trials have demonstrated that cytokine-armored CAR-T cells can overcome immunosuppression through tumor-specific cytotoxicity. However, constitutive cytokine expression introduces systemic toxicities, thereby limiting therapeutic doses. Additionally, adenosine deaminase can degrade extracellular ADO into the benign inosine metabolite, thereby relieving TME immunosuppression and restoring T cell function. These findings highlight the need for a platform that combines ADO degradation with tumor specific cytokine delivery to simultaneously resolve toxicity and efficacy barriers. Therefore, this thesis proposes an ADO-inducible, self-limiting CAR-T platform that enables tumor- localized cytokine expression in response to tumor-derived ADO. Aim 1 will define the activation threshold, dynamic range, and shutdown kinetics of the ADO-inducible cytokine

platform in ADA2Ph2 secreted HEK293T cells. Aim 2 will translate the optimized circuit into primary CAR-T cells and validate tumor-localized activation in vivo. Aim 3 will evaluate

antitumor efficacy and systemic safety in mouse breast cancer models, including in combination with immune checkpoint blockade. Together, we will establish a mechanistically defined, metabolism-guided CAR-T platform that reprograms the immunosuppressive breast cancer TME while tumor localized cytokine activity, providing a broadly applicable framework for safer and more effective breast cancer immunotherapy.