TBK1 Targeting Is Identified as a Therapeutic Strategy to Enhance CAR T-Cell Efficacy Using Patient-Derived Organotypic Tumor Spheroids

Novel therapeutic strategies are needed to improve the efficacy of chimeric antigen receptor (CAR) T cells as a treatment of solid tumors. Multiple tumor microenvironmental factors are thought to contribute to resistance to CAR T-cell therapy in solid tumors, and appropriate model systems to identify and examine these factors using clinically relevant biospecimens are limited. In this study, we examined the activity of B7-H3-directed CAR T cells (B7-H3.CAR-T) using 3D microfluidic cultures of patient-derived organotypic tumor spheroids (PDOTS) and then confirmed the activity of B7-H3.CAR T cells in PDOTS. Although B7-H3 expression in PDOTS was associated with B7-H3.CAR-T sensitivity, mechanistic studies revealed dynamic upregulation of co-inhibitory receptors on CAR T-cells following target cell encounter that led to CAR T-cell dysfunction and limited efficacy against B7-H3-expressing tumors. PD-1 blockade restored CAR T-cell activity in monotypic and organotypic tumor spheroids with improved tumor control and upregulation of effector cytokines. Given the emerging role of TANK-binding kinase 1 (TBK1) as an immune evasion gene, we examined the effect of TBK1 inhibition on CAR T-cell efficacy. Similar to PD-1 blockade, TBK1 inhibition restored CAR T-cell activity in monotypic and organotypic tumor spheroids, prevented CAR T-cell dysfunction, and enhanced CAR T-cell proliferation. Inhibition or deletion of TBK1 also enhanced the sensitivity of cancer cells to immune-mediated killing. Taken together, our results demonstrate the feasibility and utility of ex vivo profiling of CAR T cells using PDOTS and suggest that targeting TBK1 could be used to enhance CAR T-cell efficacy by overcoming tumor-intrinsic and -extrinsic resistance mechanisms.

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Publication Details

Journal
Cancer Immunology Research
Published
2025-01-09
DOI
https://doi.org/10.1158/2326-6066.cir-23-1011
Citations
13
Primary Topic
CAR-T cell therapy research
Type
article
Field-Weighted Citation Impact
13.59

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article

TBK1 Targeting Is Identified as a Therapeutic Strategy to Enhance CAR T-Cell Efficacy Using Patient-Derived Organotypic Tumor Spheroids

Arnav Mehta, Marco Ventin, Genevieve Marie Boland, Apekshya Panda et al.
13 citations
Cancer Immunology Research
CAR-T cell therapy research
13.59
article

TBK1 Targeting Is Identified as a Therapeutic Strategy to Enhance CAR T-Cell Efficacy Using Patient-Derived Organotypic Tumor Spheroids

Arnav Mehta, Marco Ventin, Genevieve Marie Boland, Apekshya Panda, Samuel J. Wright, Keith T. Flaherty, Tatyana Sharova, Amina Fu, Russell W. Jenkins, Jacy Fang, Giulia Cattaneo, Peter J. Bowling, Alex Miranda, Or‐Yam Revach, Moshe Sade-Feldman, Cristina R. Ferrone, Luke Maggs, Peter Karl Sorger, Angelina M. Cicerchia, Caitlin E. Mills, Anne Jenney, Yi Sun, Martin Q. Rasmussen, Aleigha Lawless, Xinhui Wang, Nabeel Bardeesy, Matthew D. Perricone, Feng Chen
article en
13 citations

Abstract

Novel therapeutic strategies are needed to improve the efficacy of chimeric antigen receptor (CAR) T cells as a treatment of solid tumors. Multiple tumor microenvironmental factors are thought to contribute to resistance to CAR T-cell therapy in solid tumors, and appropriate model systems to identify and examine these factors using clinically relevant biospecimens are limited. In this study, we examined the activity of B7-H3-directed CAR T cells (B7-H3.CAR-T) using 3D microfluidic cultures of patient-derived organotypic tumor spheroids (PDOTS) and then confirmed the activity of B7-H3.CAR T cells in PDOTS. Although B7-H3 expression in PDOTS was associated with B7-H3.CAR-T sensitivity, mechanistic studies revealed dynamic upregulation of co-inhibitory receptors on CAR T-cells following target cell encounter that led to CAR T-cell dysfunction and limited efficacy against B7-H3-expressing tumors. PD-1 blockade restored CAR T-cell activity in monotypic and organotypic tumor spheroids with improved tumor control and upregulation of effector cytokines. Given the emerging role of TANK-binding kinase 1 (TBK1) as an immune evasion gene, we examined the effect of TBK1 inhibition on CAR T-cell efficacy. Similar to PD-1 blockade, TBK1 inhibition restored CAR T-cell activity in monotypic and organotypic tumor spheroids, prevented CAR T-cell dysfunction, and enhanced CAR T-cell proliferation. Inhibition or deletion of TBK1 also enhanced the sensitivity of cancer cells to immune-mediated killing. Taken together, our results demonstrate the feasibility and utility of ex vivo profiling of CAR T cells using PDOTS and suggest that targeting TBK1 could be used to enhance CAR T-cell efficacy by overcoming tumor-intrinsic and -extrinsic resistance mechanisms.

Cancer Immunology ResearchVol. 13(2)
Broad Institute (US), Cedars-Sinai Medical Center (US), Harvard University (US), Massachusetts General Hospital (US), Center for Cancer Research (US), Center for Systems Biology (US)
U.S. Department of Defense, Bristol-Myers Squibb, Dr. Miriam and Sheldon G. Adelson Medical Research Foundation, Cholangiocarcinoma Foundation, Massachusetts Life Sciences Center, National Institutes of Health, National Cancer Institute, DOD Prostate Cancer Research Program
Good health and well-being
Openalex Percentile: Top 1%
CAR-T cell therapy research
13.59
DOIOpen Access PDFOpenAlex
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