Journal Articles

Ex vivo expansion of melanoma tumor infiltrating lymphocytes leads to a dominant exhausted T cell population with lack of memory markers

Tumor infiltrating lymphocytes (TILs) can be isolated from patient tumors, greatly expanded ex vivo, and returned to the patient for therapeutic effect. Recent clinical trials have highlighted the efficacy of TILs for a subset of patients and supported FDA approval for melanoma. How TILs evolve during the manufacturing process is still unknown and likely critical to improving the therapy for more patients. To characterize cell modification during TIL expansion, we performed single-cell RNA- and TCR-sequencing of TILs isolated from patient tumors and their paired ex vivo expanded cell products. We found large transcriptional differences between pre- and post-expansion TILs. Post-expansion TILs were predominantly exhausted and lacked na•ve or memory cell phenotypes, including a decreased percentage of CD39/CD69 double negative (DN) "stem-like" T cells. Co-activating receptors CD137 and CD27 decreased while CD30 increased, whereas among co-inhibitory receptors PD1 decreased while TIM3 and LAG3 showed the largest increases with expansion. Other gene families that showed large increases with ex vivo growth included cytotoxicity- and APC-associated genes. Individual clonotypes were distributed among multiple cell differentiation states, which exhibited high degrees of plasticity during expansion. Although ex vivo expanded TILs are predominantly terminally differentiated, exhausted and transcriptionally highly distinct from the initial TILs, there is also a large progenitor exhausted CD8 T cell (Tpex) population and DN numbers increase. Future work to amplify subpopulations of TILs with memory cell phenotypes, such as the DN cells, will likely further improve this therapy.

Author Info: (1) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (2) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (3) Yale University

Author Info: (1) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (2) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (3) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (4) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (5) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (6) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (7) Yale University School of Medicine New Haven, CT United States. (8) Rutgers Cancer Institute New Brunswick, NJ United States. (9) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (10) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (11) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (12) Yale University New Haven, CT United States. ROR: https://ror.org/03v76x132 (13) Yale University New Haven, Connecticut United States. ROR: https://ror.org/03v76x132

Sensitive detection of cancer antigens enabled by user-defined peptide libraries

Human leukocyte antigen (HLA)-bound tumor peptides can be routinely isolated from cancer samples and identified using mass spectrometry (MS). However, MS approaches can be stochastic or rely on spectral libraries, which are not customarily available for individual-specific peptides, thus limiting the ability to discover novel peptides. Here, we introduce Pepyrus, which generates user-defined, individual-specific or disease-specific peptide libraries in Escherichia coli to improve the sensitivity and confidence of MS peptide identification, including lowly abundant neoantigens. Using Pepyrus-generated peptide libraries paired with an HLA-specific data-independent acquisition strategy, we recover >75% of the expected sequences per single injection for libraries of >10,000 peptides and identify 0.1_fmol of spiked-in peptides in a complex background. We apply Pepyrus to create personalized libraries, facilitating identification of clinically relevant HLA peptides, including several novel peptides from cell lines derived from persons with melanoma and renal cell carcinoma. Pepyrus enables identification of rare HLA-bound peptides and provides the ability to generate large training datasets to improve spectra, retention time and ion mobility prediction tools.

Author Info: (1) Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. Dana Farber Cancer Institute, Boston, MA, USA. (2) Broad Institute of MIT and H

Author Info: (1) Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. Dana Farber Cancer Institute, Boston, MA, USA. (2) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (3) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (4) Broad Institute of MIT and Harvard, Cambridge, MA, USA. Dana Farber Cancer Institute, Boston, MA, USA. (5) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (6) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (7) Dana Farber Cancer Institute, Boston, MA, USA. (8) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (9) Dana Farber Cancer Institute, Boston, MA, USA. (10) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (11) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (12) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (13) Harvard Medical School, Boston, MA, USA. Dana Farber Cancer Institute, Boston, MA, USA. (14) Dana Farber Cancer Institute, Boston, MA, USA. (15) Department of Pathology, University of Michigan, Ann Arbor, MI, USA. (16) Dana Farber Cancer Institute, Boston, MA, USA. (17) Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. Dana Farber Cancer Institute, Boston, MA, USA. Department of Computer Science, Metropolitan College, Boston University, Boston, MA, USA. Technical University of Denmark, Lyngby, Denmark. (18) Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. Dana Farber Cancer Institute, Boston, MA, USA. (19) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (20) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (21) Broad Institute of MIT and Harvard, Cambridge, MA, USA. (22) Broad Institute of MIT and Harvard, Cambridge, MA, USA. nhacohen@broadinstitute.org. Harvard Medical School, Boston, MA, USA. nhacohen@broadinstitute.org. Massachusetts General Hospital, Krantz Family Center for Cancer Research, Boston, MA, USA. nhacohen@broadinstitute.org. (23) Broad Institute of MIT and Harvard, Cambridge, MA, USA. scarr@broad.mit.edu. (24) Broad Institute of MIT and Harvard, Cambridge, MA, USA. jabelin@broadinstitute.org. Dana Farber Cancer Institute, Boston, MA, USA. jabelin@broadinstitute.org. (25) Broad Institute of MIT and Harvard, Cambridge, MA, USA. catherine_wu@dfci.harvard.edu. Harvard Medical School, Boston, MA, USA. catherine_wu@dfci.harvard.edu. Dana Farber Cancer Institute, Boston, MA, USA. catherine_wu@dfci.harvard.edu.

Transcription factor Etv3 controls the tolerogenic function of dendritic cells

Dendritic cells (DCs) facilitate the maintenance of immunological tolerance in the steady state. We report that transcription factor Etv3 is preferentially expressed in mature DCs, including tissue-derived migratory DCs (migDCs), and facilitates their homeostatic maturation and CCR7-dependent migration. Mice with global or DC-specific deletion of Etv3 manifested the expansion of CD25(low) regulatory T (T(reg)) cells, spontaneous activation of conventional T cells, and multiorgan T cell infiltration. Etv3 deficiency exacerbated TLR7-driven systemic lupus erythematosus (SLE)-like disease, supporting the reported genetic association of human ETV3 with SLE. Etv3-deficient migDCs up-regulated multiple costimulatory molecules, including OX40 ligand (OX40L/TNFSF4), whose blockade partially rescued the T(reg) cell abnormalities. These results identify Etv3 as an essential regulator of the tolerogenic function of DCs and implicate it in the regulation of human autoimmunity.

Author Info: (1) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (2) Department of Pathology, New York University Grossman School of Medicine, New Y

Author Info: (1) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (2) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (3) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. Applied Bioinformatics Laboratories, New York University Grossman School of Medicine, New York, NY, USA. (4) Applied Bioinformatics Laboratories, New York University Grossman School of Medicine, New York, NY, USA. (5) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (6) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (7) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (8) Department of Surgery, New York University Grossman School of Medicine, New York, NY, USA. (9) Ronald O. Perelman Department of Dermatology, New York University Grossman School of Medicine, New York, NY, USA. (10) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (11) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (12) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (13) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (14) Department of Physics, Ben Gurion University of the Negev, Beer-Sheva, Israel. Ilse Katz Institute for Nanoscale Science and Technology, Ben Gurion University of the Negev, Beer-Sheva, Israel. (15) Cellular Analytics Laboratory, New York University Grossman School of Medicine, New York, NY, USA. Department of Medicine, New York University Grossman School of Medicine, New York, NY, USA. (16) Department of Medicine, New York University Grossman School of Medicine, New York, NY, USA. (17) Department of Medicine, New York University Grossman School of Medicine, New York, NY, USA. (18) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. (19) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. Ronald O. Perelman Department of Dermatology, New York University Grossman School of Medicine, New York, NY, USA. (20) Department of Pathology, New York University Grossman School of Medicine, New York, NY, USA. Department of Medicine, New York University Grossman School of Medicine, New York, NY, USA.

SLAMF6 as a drug-targetable suppressor of T cell immunity against cancer

Inhibitory receptors like PD-1 and CTLA-4 contribute to T cell dysfunction in cancer(1-3). Monoclonal antibodies (mAbs) blocking the interactions in trans of these receptors with their ligands on cancer cells or in the tumour microenvironment lead to clinical responses in some but not all types of cancer. Signalling lymphocytic activation molecule 6 (SLAMF6, also known as Ly108) is a homotypic receptor preferentially expressed on progenitor or stem-like exhausted T (T(pex)) cells, but not on terminally exhausted T (T(ex)) cells, as demonstrated in mouse models(4-9). In contrast to T(ex) cells, T(pex) cells retain the capacity for functional restoration after immune checkpoint blockade(10-12). The role of SLAMF6 in T cells remains ambiguous, as it has both activating and inhibitory effects, complicating its evaluation as a therapeutic target. Here we find that SLAMF6 was triggered in cis by homotypic interactions at the T cell surface. These interactions elicited inhibitory effects that suppressed activation of T cells and limited anti-tumour immunity, independently of SLAMF6 expression on tumour cells. mAbs against human SLAMF6 with a robust ability to disrupt the cis interactions strongly augmented T cell activation, reduced the proportions of exhausted T cells and inhibited tumour growth in vivo. Collectively, these findings show that SLAMF6 functions exclusively as a T cell inhibitory receptor, which is triggered by cis homotypic interactions. They also position SLAMF6 as a promising target for therapies aimed at enhancing anti-tumour immunity, regardless of SLAMF6 expression on tumour cells.

Author Info: (1) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. Department of Medicine, University of MontrŽal, Montreal, Quebe

Author Info: (1) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. Department of Medicine, University of MontrŽal, Montreal, Quebec, Canada. Department of Medicine, McGill University, Montreal, Quebec, Canada. (2) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. (3) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. Department of Medicine, McGill University, Montreal, Quebec, Canada. (4) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. Department of Medicine, McGill University, Montreal, Quebec, Canada. (5) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. (6) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. Cancer Center, Faculty of Health Sciences, University of Macau, Macau SAR, China. MoE Frontiers Science Center for Precision Oncology, University of Macau, Macau SAR, China. (7) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. (8) Laboratory of Molecular Oncology, Institut de recherches cliniques de MontrŽal (IRCM), Montreal, Quebec, Canada. andre.veillette@ircm.qc.ca. Department of Medicine, University of MontrŽal, Montreal, Quebec, Canada. andre.veillette@ircm.qc.ca. Department of Medicine, McGill University, Montreal, Quebec, Canada. andre.veillette@ircm.qc.ca.

Mutant KRAS vaccine with dual checkpoint blockade in resected pancreatic cancer: a phase I trial

In this phase I study, we test a pooled synthetic long peptide vaccine targeting the six KRAS mutations (G12V, G12A, G12R, G12C, G12D, G13D) with ipilimumab and nivolumab in resected pancreatic adenocarcinoma. Co-primary endpoints include safety and maximal percent change of IFN_-producing mutant KRAS T cell responses in the blood within 17 weeks. Secondary endpoints include disease-free survival, overall survival, and maximal percent change of IFN_-producing mutant KRAS T cell responses at any time after vaccination. Vaccine-related adverse events are grade 1-2. 11/12 and 10/12 patients generate a significant increase in average T cell response to 6 mutant KRAS antigens and tumor-specific response, respectively. Immunophenotyping demonstrate Th1 CD4 central memory and effector memory T cells, and CD8 effector memory T cells at a lower frequency. The vaccine also generates cross-reactive T cells that recognize more than one mutant KRAS antigen. These findings support the safety and diverse anti-tumor immunity of mutant KRAS vaccines (NCT04117087).

Author Info: (1) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins

Author Info: (1) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (2) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (3) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (4) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (5) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (6) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (7) Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (8) Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (9) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (10) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (11) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (12) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (13) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (14) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. Department of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA. (15) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (16) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (17) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (18) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. (19) Greenebaum Comprehensive Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA. (20) Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. Ludwig Center, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Lustgarten Pancreatic Cancer Research Laboratory, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Howard Hughes Medical Institute, Chevy Chase, MD, USA. Division of Rheumatology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA. (21) Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. Ludwig Center, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Lustgarten Pancreatic Cancer Research Laboratory, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Howard Hughes Medical Institute, Chevy Chase, MD, USA. Division of Rheumatology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA. (22) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (23) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (24) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (25) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (26) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (27) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (28) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (29) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (30) Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (31) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (32) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (33) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (34) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (35) Greenebaum Comprehensive Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA. (36) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (37) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. (38) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. ejaffee@jhmi.edu. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. ejaffee@jhmi.edu. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. ejaffee@jhmi.edu. (39) Johns Hopkins Convergence Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA. nzaidi1@jhmi.edu. Johns Hopkins Bloomberg Kimmel Institute for Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, USA. nzaidi1@jhmi.edu. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA. nzaidi1@jhmi.edu.

Flt3L-mediated tumor cDC1 expansion enhances immunotherapy by priming stem-like CD8+ T cells in lymph nodes

ABSTRACT: Immune checkpoint blockade (ICB) evokes antitumor immunity through the reinvigoration of T cell responses. T cell differentiation status controls response, with less differentiated cells having an enhanced capacity to proliferate after ICB. Given that conventional type 1 dendritic cells (cDC1) maintain precursor exhausted T cells (TPEX), we hypothesized that expansion of cDC1s with Flt3L could enhance responses to ICB. Here we show that treatment with Fms-related tyrosine kinase 3 ligand (Flt3L) expands CD62L+SLAMF6+CD8+ T cells in the tumor through a mechanism that requires XCR1+ dendritic cells to traffic to the tumor-draining lymph node. The combination of Flt3L and anti-CTLA-4 enhanced therapeutic responses. Combination therapy is associated with the emergence of a CD8+ T cell subset characterized by the expression of Il21r and oligoclonal expansion of CD8+ T cells within tumors through a mechanism that is dependent on lymph node egress.

Author Info: (1) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Vi

Author Info: (1) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (2) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (3) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. School of Biomedical Sciences, The University of Western Australia, Perth, Western Australia, Australia. The Kids Research Institute Australia, The University of Western Australia, Perth, Western Australia, Australia. (4) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (5) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (6) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (7) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (8) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (9) Harry Perkins Institute of Medical Research, QEII Medical Centre and Centre for Medical Research, The University of Western Australia, Perth, Western Australia, Australia. (10) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (11) Institute for Respiratory Health, National Centre for Asbestos Related Diseases, The University of Western Australia, Perth, Western Australia, Australia. (12) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (13) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (14) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (15) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (16) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (17) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (18) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (19) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (20) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (21) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (22) Department of Microbiology and Immunology at the Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Parkville, Victoria, Australia. (23) Institute for Respiratory Health, National Centre for Asbestos Related Diseases, The University of Western Australia, Perth, Western Australia, Australia. (24) Department of Microbiology and Immunology at the Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Parkville, Victoria, Australia. (25) Harry Perkins Institute of Medical Research, QEII Medical Centre and Centre for Medical Research, The University of Western Australia, Perth, Western Australia, Australia. (26) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. (27) Asthma and Airway Disease Research Center, University of Arizona, Tucson, AZ, USA. Department of Immunobiology, The University of Arizona College of Medicine, Tucson, AZ, USA. (28) School of Biomedical Sciences, The University of Western Australia, Perth, Western Australia, Australia. jason.waithman@uwa.edu.au. (29) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. paul.beavis@petermac.org. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. paul.beavis@petermac.org. (30) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia. phil.darcy@petermac.org. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia. phil.darcy@petermac.org. Department of Immunology, Monash University, Clayton, Victoria, Australia. phil.darcy@petermac.org.

Persistent T cell activation and cytotoxicity against glioblastoma following single oncolytic virus treatment in a clinical trial Featured  

Following clinical evidence that a single oncolytic virus treatment was associated with immune activation signatures, Meylen, Tiian, Wu, Ling, et al. analyzed tumor samples and found that pre-existing TILs expanded upon treatment, resulting in deep and persistent T cell activation against tumor cells. While viral remnants were restricted to necrotic regions, granzyme B+ CD8+ T cells embedded deeply into tumors, showed persistent activation, and were located in close proximity to apoptotic tumor cells. These T cell observations further and that this correlated with longer-progression-free and overall survival.

Following clinical evidence that a single oncolytic virus treatment was associated with immune activation signatures, Meylen, Tiian, Wu, Ling, et al. analyzed tumor samples and found that pre-existing TILs expanded upon treatment, resulting in deep and persistent T cell activation against tumor cells. While viral remnants were restricted to necrotic regions, granzyme B+ CD8+ T cells embedded deeply into tumors, showed persistent activation, and were located in close proximity to apoptotic tumor cells. These T cell observations further and that this correlated with longer-progression-free and overall survival.

ABSTRACT: A recent first-in-human clinical trial demonstrated that survival in glioblastoma (GBM) patients following rQNestin34.5v.2 oncolytic virus treatment was associated with immune activation signatures. This study was registered at ClinicalTrials.gov (NCT03152318). Here, we provide in situ evidence of ongoing T cell-mediated cytotoxicity against tumor cells at late time points following single treatment, with deep and persistent T cell infiltration into tumor regions. Shorter distances between cleaved caspase-3(+) tumor cells and granzyme B(+) T cells were associated with longer progression-free survival following treatment. Pre-existing tumor-infiltrating T cells expanded locally upon treatment, correlating with longer overall patient survival. T cells with an early activation program closely interacted with tumor cells and were strongly enriched upon treatment. Viral remnants were restricted to necrotic regions, while T cells infiltrated deeply into live tumor regions. These data demonstrate that single oncolytic virus treatment can expand pre-existing T cell clones and trigger persistent T cell-mediated immunity against GBM.

Author Info: (1) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Immunology, Harvard Medical School, Boston, MA, USA. (2) Department o

Author Info: (1) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Immunology, Harvard Medical School, Boston, MA, USA. (2) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Immunology, Harvard Medical School, Boston, MA, USA. (3) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Immunology, Harvard Medical School, Boston, MA, USA. (4) Harvey Cushing Neuro-Oncology Laboratories, Department of Neurosurgery and Center for Tumors of the Nervous System, Neuroscience Institute and Cancer Institute, Mass General Brigham, Boston, MA, USA. (5) Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel. (6) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Immunology, Harvard Medical School, Boston, MA, USA. (7) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA. (8) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA. (9) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Immunology, Harvard Medical School, Boston, MA, USA. (10) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA. (11) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM), Heidelberg, Germany; German Cancer Research Center (DKFZ), JRG Hematology and Immune Engineering, Heidelberg, Germany. (12) Department of Pathology and Krantz Family Center for Cancer Research, Massachusetts General Hospital, Boston, MA, USA; Broad Institute of MIT and Harvard, Cambridge, MA, USA. (13) Department of Pathology and Krantz Family Center for Cancer Research, Massachusetts General Hospital, Boston, MA, USA; Broad Institute of MIT and Harvard, Cambridge, MA, USA. (14) Harvey Cushing Neuro-Oncology Laboratories, Department of Neurosurgery and Center for Tumors of the Nervous System, Neuroscience Institute and Cancer Institute, Mass General Brigham, Boston, MA, USA. (15) Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel. (16) Department of Pathology and Krantz Family Center for Cancer Research, Massachusetts General Hospital, Boston, MA, USA; Broad Institute of MIT and Harvard, Cambridge, MA, USA. Electronic address: suva.mario@mgh.harvard.edu. (17) Harvey Cushing Neuro-Oncology Laboratories, Department of Neurosurgery and Center for Tumors of the Nervous System, Neuroscience Institute and Cancer Institute, Mass General Brigham, Boston, MA, USA. Electronic address: eachiocca@mgb.org. (18) Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, USA; Department of Immunology, Harvard Medical School, Boston, MA, USA; Department of Neurology, Brigham and Women's Hospital, Boston, MA, USA. Electronic address: kai_wucherpfennig@dfci.harvard.edu.

TNF-⍺-mediated myeloid-instructed CD14+CD4+ T cells are associated with poor survival in lung adenocarcinoma

Spotlight 

Marceaux et al. performed spatial multi-omics analysis across clinically annotated LUAD and LUSC cohorts of NSCLC, and stratified it into lymphoid-, myeloid-, and mixed-enriched subtypes, independent of driver mutations. CD14⁺CD4⁺ T cells were enriched in myeloid niches and absent in healthy lungs. CD4+ T cells acquired CD14 from HLA-DR⁺ myeloid cells via trogocytosis, forming an atypical myeloid-instructed phenotype. TNFα signaling was enriched in CD14+CD4+ T cell-high tumors. The addition of TNF⍺ enhanced trogocytosis and CD14+CD4+ T cells formation ex vivo. CD14+CD4+ T cells represent a distinct T cell population associated with poor prognosis in NSCLC.

Contributed by Shishir Pant

Marceaux et al. performed spatial multi-omics analysis across clinically annotated LUAD and LUSC cohorts of NSCLC, and stratified it into lymphoid-, myeloid-, and mixed-enriched subtypes, independent of driver mutations. CD14⁺CD4⁺ T cells were enriched in myeloid niches and absent in healthy lungs. CD4+ T cells acquired CD14 from HLA-DR⁺ myeloid cells via trogocytosis, forming an atypical myeloid-instructed phenotype. TNFα signaling was enriched in CD14+CD4+ T cell-high tumors. The addition of TNF⍺ enhanced trogocytosis and CD14+CD4+ T cells formation ex vivo. CD14+CD4+ T cells represent a distinct T cell population associated with poor prognosis in NSCLC.

Contributed by Shishir Pant

ABSTRACT: The tumor microenvironment is composed of diverse immune populations that can either support anti-tumor immunity or promote tumor progression. Myeloid cells are major drivers of immunosuppression, yet therapies targeting them have shown limited success. To uncover mechanisms underlying myeloid-driven immune suppression, we performed spatial multi-omics analyses of non-small cell lung cancer (NSCLC). Independent of oncogenic driver status, tumors stratify into lymphoid-enriched, myeloid-enriched, and mixed immune-infiltrated subtypes. In tumor and adjacent non-malignant lungs, we identify myeloid-instructed CD14+CD4+ T cells. These cells arise through trogocytosis adopting an atypical phenotype. In lymphoid-enriched tumors, high infiltration of CD14+CD4+ T cells correlates with poor patient survival. Spatial transcriptomics reveal enrichment of tumor necrosis factor alpha (TNF-α) signaling in CD14+CD4+-T-cell-rich tumors. Functional assays demonstrate that TNF-⍺ enhanced trogocytosis, promoting the formation of CD14+CD4+ T cells. These findings uncover a TNF-⍺-mediated mechanism of immunosuppression in the TME and highlight aberrant myeloid-T cell interactions as contributors to NSCLC progression.

Author Info: (1) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; Department of Medical Biology, The University of Mebourne, P

Author Info: (1) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; Department of Medical Biology, The University of Mebourne, Parkville, VIC, Australia. (2) Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (3) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (4) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (5) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (6) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; Department of Medical Biology, The University of Mebourne, Parkville, VIC, Australia. (7) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; Department of Medical Biology, The University of Mebourne, Parkville, VIC, Australia. (8) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (9) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (10) Advanced Technology and Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (11) Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. (12) The Royal Melbourne Hospital, Parkville, VIC, Australia. (13) The Royal Melbourne Hospital, Parkville, VIC, Australia. (14) Department of Surgery, The University of Melbourne, Parkville, VIC, Australia; The Royal Melbourne Hospital, Parkville, VIC, Australia. (15) Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; School of Mathematics and Statistics, The University of Melbourne, Parkville, VIC, Australia. (16) Advanced Technology and Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; Department of Medical Biology, The University of Mebourne, Parkville, VIC, Australia. (17) Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; Department of Medical Biology, The University of Mebourne, Parkville, VIC, Australia. (18) Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; Department of Medical Biology, The University of Mebourne, Parkville, VIC, Australia. Electronic address: labat@wehi.edu.au.

Chimeric antigen receptor T cells against the IGHV4-34 B cell receptor specifically eliminate neoplastic and autoimmune B cells Spotlight 

Focused on key challenges of CD19 CAR T cell therapy, such as on-target/off-tumor healthy B cell depletion and antigen-negative escape and relapse, Cohen, Bochi-­Layec and Lemoine et al. developed CAR T cells targeting the BCR-carrying IGHV-34 (CART4-34), which is highly enriched in B cell cancers, SLE, and other autoimmune diseases. CART4-34 potently killed malignant B cells and pathogenic B cells in SLE ex vivo, while sparing normal IGHV-34-negative B cells. In preclinical models, CART4-­34 showed robust expansion and antitumor activity, comparable to CART19, but was associated with reduced antigen-negative escape following treatment.

Contributed by Katherine Turner

Focused on key challenges of CD19 CAR T cell therapy, such as on-target/off-tumor healthy B cell depletion and antigen-negative escape and relapse, Cohen, Bochi-­Layec and Lemoine et al. developed CAR T cells targeting the BCR-carrying IGHV-34 (CART4-34), which is highly enriched in B cell cancers, SLE, and other autoimmune diseases. CART4-34 potently killed malignant B cells and pathogenic B cells in SLE ex vivo, while sparing normal IGHV-34-negative B cells. In preclinical models, CART4-­34 showed robust expansion and antitumor activity, comparable to CART19, but was associated with reduced antigen-negative escape following treatment.

Contributed by Katherine Turner

ABSTRACT: Current US Food and Drug Administration-approved chimeric antigen receptor (CAR) T cell therapies for B cell leukemias and lymphomas target CD19, which is widely expressed across the B cell lineage, often leading to on-target, off-tumor B cell depletion, prolonged immune suppression, and antigen-negative escape in a subset of patients. In contrast, B cell receptor (BcR) signaling is essential for the survival of most mature B cell neoplasms, and BcRs carrying the immunoglobulin heavy variable gene IGHV4-34 are highly enriched in B cell malignancies compared with normal B cells. Further, self-reactive IGHV4-34(+) serum autoantibodies are enriched in aggressive systemic lupus erythematosus (SLE) and other autoimmune diseases. Here, we developed CAR T cells targeting the BcR carrying IGHV4-34 (CART4-34). We found that CART4-34 showed specific cytotoxicity and cytokine secretion toward IGHV4-34(+) malignant B cells. In addition, although CD19 was down-regulated upon relapse after treatment with CART19, IGHV4-34(+) BcR levels remained intact upon relapse after treatment with CART4-34, suggesting reduced risk of antigen-negative escape. In IGHV4-34(+) HBL1 cell line-derived xenograft mouse models, CART4-34 showed robust expansion and antitumor activity comparable to those of CART19. Optimized CAR:BcR binding using shorter CAR hinge domains improved immune synapse morphology and in vivo activity. In addition, we showed that CART4-34 could target human IGHV4-34(+) SLE B cells and deplete IGHV4-34(+) autoantibodies ex vivo, without targeting healthy B cells or affecting total IgG titers. In conclusion, we developed a CAR T cell product that specifically targets pathogenic B cells in lymphoid malignancies and SLE, offering potential for precision cell therapy for these indications.

Author Info: (1) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia

Author Info: (1) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (2) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (3) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (4) Department of Medicine, Division of Rheumatology, School of Medicine, Lowance Center for Human Immunology, Emory University, Atlanta, GA 30322, USA. (5) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (6) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (7) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Surgery, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (8) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (9) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (10) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (11) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (12) Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA. (13) Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki 57001, Greece. (14) Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki 57001, Greece. (15) Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki 57001, Greece. (16) Division of Rheumatology, Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. (17) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (18) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (19) Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki 57001, Greece. Division of Genetics and Biotechnology, Department of Biology, National and Kapodistrian University of Athens, Athens 15772, Greece. (20) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (21) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (22) Division of Hematopathology, Department of Pathology and Laboratory Medicine, Weill Cornell Medicine/New York-Presbyterian Hospital, New York, NY 10065, USA. (23) LatchBio, San Francisco, CA 94107, USA. (24) Department of Medicine, Division of Rheumatology, School of Medicine, Lowance Center for Human Immunology, Emory University, Atlanta, GA 30322, USA. (25) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (26) Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine and Therapeutic Pathology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (27) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine and Therapeutic Pathology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (28) Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (29) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (30) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (31) Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA. (32) Laboratory of Cancer Biology and Genetics, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (33) Cancer Data Science Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (34) Cancer Data Science Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (35) Institute of Hematology and Center for Hemato-Oncology Research, Department of Medicine and Surgery, University and Hospital of Perugia, Perugia 06129, Italy. (36) Institute for Immunology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA. (37) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine and Therapeutic Pathology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (38) Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. (39) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (40) Institute for Immunology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA. (41) Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (42) Institute of Hematology and Center for Hemato-Oncology Research, Department of Medicine and Surgery, University and Hospital of Perugia, Perugia 06129, Italy. (43) Division of Rheumatology, Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. (44) Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA. (45) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine and Therapeutic Pathology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (46) Department of Medicine, Division of Rheumatology, School of Medicine, Lowance Center for Human Immunology, Emory University, Atlanta, GA 30322, USA. (47) Division of Hematopathology, Department of Pathology and Laboratory Medicine, Weill Cornell Medicine/New York-Presbyterian Hospital, New York, NY 10065, USA. (48) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA. (49) Medical School, Universitˆ Vita-Salute San Raffaele, Milan 20132, Italy. B cell Neoplasia Unit, Comprehensive Cancer Center, IRCCS Ospedale San Raffaele, Milan 20132, Italy. (50) Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki 57001, Greece. (51) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, PA 19104, USA. Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Division of Hematology/Oncology, Hospital of University of Pennsylvania, Philadelphia, PA 19104, USA.

Combined targeted and epigenetic-based therapy enhances antitumor immunity by stabilizing GATA6-dependent MHCI expression in pancreatic ductal adenocarcinoma Spotlight 

Peng, Yang, Antonopoulou, et al. found that in human PDAC, high expression of GATA6 correlated with increased MHC-I expression, immune cell infiltration, and interactions with CD8+ T cells. In murine tumor lines, MEK inhibition (MEKi) further increased MHC-I expression in GATA6high tumor cells, leading to enhanced T cell cytotoxicity against them, while GATA6 knockout or degradation abrogated this effect. In vivo, high GATA6 expression was required for MEKi-induced tumor control, but long-term treatment reduced GATA6+ cells and increased immunosuppressive EMT, which could be overcome by combining MEKi with HDAC inhibitors.

Contributed by Lauren Hitchings

Peng, Yang, Antonopoulou, et al. found that in human PDAC, high expression of GATA6 correlated with increased MHC-I expression, immune cell infiltration, and interactions with CD8+ T cells. In murine tumor lines, MEK inhibition (MEKi) further increased MHC-I expression in GATA6high tumor cells, leading to enhanced T cell cytotoxicity against them, while GATA6 knockout or degradation abrogated this effect. In vivo, high GATA6 expression was required for MEKi-induced tumor control, but long-term treatment reduced GATA6+ cells and increased immunosuppressive EMT, which could be overcome by combining MEKi with HDAC inhibitors.

Contributed by Lauren Hitchings

ABSTRACT: GATA6 promotes epithelial phenotypes and limits epithelial-to-mesenchymal (EMT) transition in pancreatic ductal adenocarcinoma (PDAC). Here we show that GATA6 defines a tumor cell state that induces MHCI expression and anti-tumor cytotoxicity upon therapy. In human PDAC, GATA6 expression correlates with immune cell infiltration, and spatial analysis reveals interaction between GATA6(+) tumor cells and CD8(+) T cells. In murine PDAC, MEK inhibition (MEKi) enriches antigenicity-related gene sets in GATA6(high) cells, while GATA6 knockout or degradation impairs MEKi-induced MHCI upregulation. High-GATA6 tumors respond to MEKi with increased MHCI, enhancing T-cell cytotoxicity, whereas GATA6 loss abolishes this effect. Treatment-induced EMT reduces GATA6(+) populations and MHCI expression, which is restored by combining MEKi with HDAC inhibitors, enhancing GATA6(+) tumor cells, MHCI, CD8(+) T cell infiltration, tumor suppression, and survival. These findings suggest that therapeutic strategies promoting a GATA6-driven tumor cell state improve immune recognition of PDAC cells and potentiate anti-tumor cytotoxic effects.

Author Info: (1) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University

Author Info: (1) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. Department of Gastroenterology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. (2) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (3) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (4) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (5) Institute of Biochemistry, University of Kiel, Kiel, Germany. (6) Institute of Biochemistry, University of Kiel, Kiel, Germany. (7) Institute of Biochemistry, University of Kiel, Kiel, Germany. (8) Division Immune Regulation in Cancer, German Cancer Research Center (DKFZ) Heidelberg, Heidelberg, Germany. (9) Division Immune Regulation in Cancer, German Cancer Research Center (DKFZ) Heidelberg, Heidelberg, Germany. (10) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (11) Department of Internal Medicine II, Klinikum rechts der Isar der Technischen UniversitŠt MŸnchen, Munich, Germany. (12) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (13) Department of Internal Medicine II, Klinikum rechts der Isar der Technischen UniversitŠt MŸnchen, Munich, Germany. (14) German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. Department of Urology, West German Cancer Center, University Hospital Essen, Essen, Germany. Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada. (15) German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. Department of Urology, West German Cancer Center, University Hospital Essen, Essen, Germany. Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada. (16) Institute of Pathology, School of Medicine and Health, Technical University of Munich, Munich, Germany. German Cancer Consortium (DKTK), Partner Site Munich, Munich, Germany. (17) Institute of Pathology, School of Medicine and Health, Technical University of Munich, Munich, Germany. German Cancer Consortium (DKTK), Partner Site Munich, Munich, Germany. (18) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (19) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (20) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (21) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (22) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. (23) German Cancer Consortium (DKTK), Partner Site Munich, Munich, Germany. School of Medicine and Health, Klinikum rechts der Isar, Technical University of Munich (TUM), Munich, Germany, Department of Diagnostic and Interventional Radiology and Department of Nuclear Medicine, University Medical Center Hamburg Eppendorf, Hamburg, Germany. (24) Department of Engineering for Innovation Medicine, University of Verona, Verona, Italy. ARC-Net Research Centre, University and Hospital Trust of Verona, Verona, Italy. (25) ARC-Net Research Centre, University and Hospital Trust of Verona, Verona, Italy. Department of Diagnostics and Public Health, University of Verona, Verona, Italy. (26) EPO - Experimental Pharmacology and Oncology GmbH, Berlin, Germany. (27) Institute of Immunology, Medical Faculty, University of Duisburg-Essen, Essen, Germany. (28) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. f.cheung@dkfz-heidelberg.de. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. f.cheung@dkfz-heidelberg.de. Spatiotemporal tumor heterogeneity, DKTK, partner site Essen, a partnership between DKFZ and University Hospital Essen, Essen, Germany. f.cheung@dkfz-heidelberg.de. (29) Bridge Institute of Experimental Tumor Therapy (BIT) and Division of Solid Tumor Translational Oncology (DKTK), West German Cancer Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. j.siveke@dkfz.de. German Cancer Consortium (DKTK), partner site Essen, a partnership between German Cancer Research Center (DKFZ) and University Hospital Essen, Essen, Germany. j.siveke@dkfz.de. National Center for Tumor Diseases (NCT) West, Campus Essen, Essen, Germany. j.siveke@dkfz.de.

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