2. Academic Validation
  3. Lymphotoxin-driven cancer cell eradication by tumoricidal CD8+ TIL

Lymphotoxin-driven cancer cell eradication by tumoricidal CD8+ TIL

  • bioRxiv. 2025 Nov 20:2025.11.19.689204. doi: 10.1101/2025.11.19.689204.
Hongyan Xie 1 2 3,  Aiping Jiang 1 2 3,  Aonkon Dey 1 2 3 4,  Joseph W Dean 5,  Jonathan J Perera 3,  Neal P Smith 1 2 3 4,  Alex C Y Chen 1 2 3 4,  Seth Anderson 1 3,  Angelina M Cicerchia 1,  Yi Sun 1,  William A Michaud 6,  Maria Florentin 1,  Jacy Fang 1 3,  Or-Yam Revach 1 2 3,  Tatyana Sharova 6,  Aleigha Lawless 6,  Katherine H Xu 1,  Yuhui Song 1,  Bidish Patel 1,  Jonathan D Stevens 7,  William J Lane 7,  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 Gastman 5,  Rongsu Qi 5,  Hequn Yin 5,  Alexandra-Chloé Villani 1 2 3 4,  Robert T Manguso 1 2 3,  Russell W Jenkins 1 2 3 12 13
Affiliations

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.
PMID: 41332750 DOI: 10.1101/2025.11.19.689204
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.

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