Lymphotoxin-driven cancer cell eradication by tumoricidal CD8+ TIL

Hongyan Xie; Aiping Jiang; Aonkon Dey; Joseph W Dean; Jonathan J Perera; Neal P Smith; Alex C Y Chen; Seth Anderson; Angelina M Cicerchia; Yi Sun; William A Michaud; Maria Florentin; Jacy Fang; Or-Yam Revach; Tatyana Sharova; Aleigha Lawless; Katherine H Xu; Yuhui Song; Bidish Patel; Jonathan D Stevens; William J Lane; Derin B Keskin; Sonia Cohen; Donald P Lawrence; Ryan J Sullivan; Keith T Flaherty; Genevieve M Boland; Linda T Nieman; Moshe Sade-Feldman; Nir Hacohen; Debattama R Sen; Catherine Wu; Brian Gastman; Rongsu Qi; Hequn Yin; Alexandra-Chloé Villani; Robert T Manguso; Russell W Jenkins

doi:10.1101/2025.11.19.689204

Lymphotoxin-driven cancer cell eradication by tumoricidal CD8⁺ TIL

bioRxiv [Preprint]. 2025 Nov 20:2025.11.19.689204. doi: 10.1101/2025.11.19.689204.

Authors

Hongyan Xie^{1

2

3}, Aiping Jiang^{1

2

3}, Aonkon Dey^{1

2

3

4}, Joseph W Dean⁵, Jonathan J Perera³, Neal P Smith^{1

2

3

4}, Alex C Y Chen^{1

2

3

4}, Seth Anderson^{1

3}, Angelina M Cicerchia¹, Yi Sun¹, William A Michaud⁶, Maria Florentin¹, Jacy Fang^{1

3}, Or-Yam Revach^{1

2

3}, Tatyana Sharova⁶, Aleigha Lawless⁶, Katherine H Xu¹, Yuhui Song¹, Bidish Patel¹, Jonathan D Stevens⁷, William J Lane⁷, Derin B Keskin^{2

3

8

9

10

11}, Sonia Cohen^{1

2

6}, Donald P Lawrence^{2

12}, Ryan J Sullivan^{2

12}, Keith T Flaherty^{2

12}, Genevieve M Boland^{1

2

6}, Linda T Nieman^{1

2}, Moshe Sade-Feldman^{1

2

3}, Nir Hacohen^{1

2

3}, Debattama R Sen^{1

2

3}, Catherine Wu^{2

9}, Brian Gastman⁵, Rongsu Qi⁵, Hequn Yin⁵, Alexandra-Chloé Villani^{1

2

3

4}, Robert T Manguso^{1

2

3}, Russell W Jenkins^{1

2

3

12

13}

Affiliations

¹ Mass General Cancer Center, Krantz Family Center for Cancer Research, Department of Medicine, Massachusetts General Hospital, Boston, MA, USA.
² Harvard Medical School, Boston, MA, USA.
³ Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁴ Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Boston, MA, USA.
⁵ Iovance Biotherapeutics, Inc., San Carlos, CA, USA.
⁶ Division of Gastrointestinal and Oncologic Surgery, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA.
⁷ Department of Pathology, Brigham and Women's Hospital, Boston, MA 02215, USA.
⁸ Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
⁹ Translational Immunogenomics Laboratory, Dana-Farber Cancer Institute, Boston, Massachusetts, USA.
¹⁰ Department of Computer Science, Metropolitan College, Boston University, Boston, Massachusetts, USA.
¹¹ Section for Bioinformatics, Department of Health Technology, Technical University of Denmark, Lyngby, Denmark.
¹² Mass General Cancer Center, Department of Medicine, Massachusetts General Hospital, Boston, MA, USA.
¹³ Present address: Hollings Cancer Center, Medical University of South Carolina, Charleston, South Carolina, USA.

Abstract

Tumor-infiltrating lymphocyte (TIL) therapy is FDA-approved for patients with treatment-resistant advanced melanoma, but the TIL subpopulations critical for tumor eradication remains incompletely understood. Using patient-derived TIL-melanoma co-cultures, we identified and characterized a novel subset of CD8⁺ TIL, capable of class I HLA-independent cancer cell lysis. The lymphotoxin β receptor (LTβR) and interferon (IFN) sensing pathways were nominated as key determinants of TIL-mediated cancer cell killing from a whole-genome, loss-of-function CRISPR screen. Validation studies confirmed that dual LTβR and IFN sensing is necessary and sufficient for cancer cell lysis, and that expanded CD8⁺ TIL express high lymphotoxin β (LTB) and upregulate lymphotoxin α (LTA) upon coculture with cancer cells. Leveraging paired scRNA-seq and scTCR-seq data, we confirmed that enrichment of LTB ⁺ CD8 ⁺ T cells is associated with clinical response to TIL, and that LTB ⁺ CD8 ⁺ TIL are expanded from putative neoantigen-reactive, LTB ^lo CD8⁺ T cells in resected tumors.

Keywords: cell death; interferon; lymphotoxin; melanoma; tumor-infiltrating lymphocytes.

Publication types

Preprint

Abstract

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