Bridging Bench to Bedside: Developing Sup2-IL-33 armored CAR T cells for translational immunotherapy
Author ORCID Identifier
https://orcid.org/0000-0002-1580-9289
Date of Award
10-13-2025
Document Type
Thesis (Ph.D.)
Department or Program
Microbiology and Immunology
First Advisor
Yina Huang
Abstract
Despite some promising results, CAR T cell therapy elicits limited responses in most patients with solid tumors. Armoring CAR T cells with immunomodulatory cytokines improves efficacy in solid tumors. We previously reported that administering IL-2 superkine (Sup2) and IL-33 cytokine armored CAR T cells without host pre-conditioning is sufficient to broadly activate host immunity to delay growth of B16F10 melanoma in immunocompetent B6 mice. However, lymphodepleting pre-conditioning prior to CAR T cell infusion is routinely utilized to enable engraftment of adoptively transferred cells. Here we assess the efficacy TRP1-specific CAR T cells with or without Sup2 and IL-33 armoring following local or systemic pre-conditioning regimens. CAR T cell expansion kinetics and phenotype were evaluated with luminescence imaging, flow cytometry, and single-cell RNA sequencing. We found that systemically pre-conditioned mice treated with Sup2-IL33 CAR T cells exerted substantially greater tumor control and extended overall survival compared to non- or locally pre-conditioned mice. Pre-conditioning with total body irradiation (TBI) increased expression of IFN stimulated genes promoting the emergence of novel Tox- effector phenotypes following Sup2-IL33 CAR T cell treatment. Durable responsiveness and extended survival following combined therapy required continued CAR T cell persistence despite the ability of Sup2-IL33 armoring to induce endogenous tumor immunity.
Limitations in mouse tumor models can hinder clinical translation of novel immunotherapeutic treatments. Bone, liver, and thymus (BLT) humanized mice recapitulate many components of the human immune system, allowing pre-clinical therapies to be assessed in the context of human immune cells during development. We evaluated the efficacy of Sup2-IL33 armored TIM-1 specific CAR T cells against Caki-1 renal cell carcinoma tumors in BLT humanized mice. Human immune reconstitution, tumor antigen expression, and CAR T cell phenotype were evaluated with flow cytometry and luminescence imaging. Despite effective in vitro killing of Caki-1 tumor cells, Sup2-IL33 CAR T cells did not improve in vivo tumor control. However, cytokine armoring increased the expansion of adoptively transferred CD8 T cells, suggesting that Sup2-IL33 armoring may be beneficial for proliferation of human T cells. Together, these studies aim to advance Sup2-IL33 CAR T cell therapy through evaluation in more clinically relevant models.
Recommended Citation
Ferry, Shannon Lynn, "Bridging Bench to Bedside: Developing Sup2-IL-33 armored CAR T cells for translational immunotherapy" (2025). Dartmouth College Ph.D Dissertations. 433.
https://digitalcommons.dartmouth.edu/dissertations/433
