T cells genetically engineered to overcome death signaling enhance adoptive cancer immunotherapy
Tori N. Yamamoto, … , Nicholas P. Restifo, Christopher A. Klebanoff
Tori N. Yamamoto, … , Nicholas P. Restifo, Christopher A. Klebanoff
Published January 29, 2019
Citation Information: J Clin Invest. 2019;129(4):1551-1565. https://doi.org/10.1172/JCI121491.
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Research Article Immunology Oncology

T cells genetically engineered to overcome death signaling enhance adoptive cancer immunotherapy

Abstract

Across clinical trials, T cell expansion and persistence following adoptive cell transfer (ACT) have correlated with superior patient outcomes. Herein, we undertook a pan-cancer analysis to identify actionable ligand-receptor pairs capable of compromising T cell durability following ACT. We discovered that FASLG, the gene encoding the apoptosis-inducing ligand FasL, is overexpressed within the majority of human tumor microenvironments (TMEs). Further, we uncovered that Fas, the receptor for FasL, is highly expressed on patient-derived T cells used for clinical ACT. We hypothesized that a cognate Fas-FasL interaction within the TME might limit both T cell persistence and antitumor efficacy. We discovered that genetic engineering of Fas variants impaired in the ability to bind FADD functioned as dominant negative receptors (DNRs), preventing FasL-induced apoptosis in Fas-competent T cells. T cells coengineered with a Fas DNR and either a T cell receptor or chimeric antigen receptor exhibited enhanced persistence following ACT, resulting in superior antitumor efficacy against established solid and hematologic cancers. Despite increased longevity, Fas DNR–engineered T cells did not undergo aberrant expansion or mediate autoimmunity. Thus, T cell–intrinsic disruption of Fas signaling through genetic engineering represents a potentially universal strategy to enhance ACT efficacy across a broad range of human malignancies.

Authors

Tori N. Yamamoto, Ping-Hsien Lee, Suman K. Vodnala, Devikala Gurusamy, Rigel J. Kishton, Zhiya Yu, Arash Eidizadeh, Robert Eil, Jessica Fioravanti, Luca Gattinoni, James N. Kochenderfer, Terry J. Fry, Bulent Arman Aksoy, Jeffrey E. Hammerbacher, Anthony C. Cruz, Richard M. Siegel, Nicholas P. Restifo, Christopher A. Klebanoff

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Figure 6

Expression of Fas DNR enhances antiapoptotic functions and in vivo persistence in anti-CD19 CAR model.

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Expression of Fas DNR enhances antiapoptotic functions and in vivo persi...
(A) Representative flow plots and (B) summary data of double transduction of B6 CD8α+ T cells with retroviral constructs encoding anti-CD19 CAR and empty or Fas DNR. Analysis performed on day 11 after Thy1.1 bead enrichment on day 6. (C) Summary bar graph of relative T cell viability (to FasΔDD) following overnight culture in cytokine-free media alone, with lz-FasL (100 ng ml–1), 2 μg ml–1 each of anti-CD3 and anti-CD28, or E2a-PBX. Data shown after gating on Thy1.1+ lymphocytes are representative of 3 independently performed experiments, and displayed as mean ± SEM with n = 3 per condition. *P < 0.05, ****P < 0.0001, 2-way ANOVA. (D) Experimental schema for the generation and infusion of WT CD8α+ T cells engineered to express anti-CD19 CAR along with FasΔDD DNR or an empty vector control. Transduced T cells were Thy1.1 bead enriched prior to injection, and T cells were infused i.v. into sublethally irradiated (5 Gy) mice bearing 4-day-established E2a-PBX leukemia. Spleens and BM were harvested for analysis on day 14. co-Td, cotransduced. (E) Summary data of numbers of live CD8α+Thy1.1+ lymphocytes in spleens and BM of recipient mice. (F) Summary data of the frequency of E2a-PBX leukemia in the BM of recipient mice. Results in E and F are representative of 2 independent experiments, each with n = 3–5 mice. *P < 0.05, **P < 0.01, ****P < 0.0001, 1-way ANOVA, corrected with Tukey’s multiple comparisons. (G) Survival of mice bearing 4-day-established E2a-PBX leukemia that were untreated or received 3 × 105 (left) or 2 × 105 (right) anti-CD19 CAR+ Thy1.1+ modified cells. Representative results from 4 independent experiments are shown as mean ± SEM using n = 5 mice/cohort. Statistical comparisons were performed using the log-rank Mantel-Cox test; *P < 0.05 **P < 0.01.