Journal Articles

FLT3L-secreting cDC1 in situ vaccination enhances antitumor immunity and synergizes with PD-1 blockade in murine non-small cell lung cancer Spotlight 

Abascal et al. engineered FLT3L-secreting mouse cDC1s that retained APC and phagocytic ability in vitro. In situ vaccination (ISV) with the cDC1s inhibited s.c. tumor growth in multiple syngeneic murine models, including those with driver mutations common in human NSCLC, and increased trafficking of the autologous cDC1s to TdLN and tumor infiltration of T cells. TCGA analysis showed that FLT3L expression in human NSCLC correlated with profiles of B and T cells, activated DCs and HEV-enriched TLS. ISV increased immature TLS formation in the murine TIME and synergized with anti-PD-1 in a NSCLC model to enhance efficacy and induce immune memory.

Contributed by Paula Hochman

Abascal et al. engineered FLT3L-secreting mouse cDC1s that retained APC and phagocytic ability in vitro. In situ vaccination (ISV) with the cDC1s inhibited s.c. tumor growth in multiple syngeneic murine models, including those with driver mutations common in human NSCLC, and increased trafficking of the autologous cDC1s to TdLN and tumor infiltration of T cells. TCGA analysis showed that FLT3L expression in human NSCLC correlated with profiles of B and T cells, activated DCs and HEV-enriched TLS. ISV increased immature TLS formation in the murine TIME and synergized with anti-PD-1 in a NSCLC model to enhance efficacy and induce immune memory.

Contributed by Paula Hochman

BACKGROUND: Non-small cell lung cancer (NSCLC) frequently evades immune surveillance through defective antigen presentation and a suppressive tumor microenvironment (TME), limiting the efficacy of immune checkpoint blockade (ICB). Conventional type 1 dendritic cells (cDC1s) are essential for initiating antitumor CD8(+) T-cell responses; however, their abundance and function are often diminished in NSCLC, contributing to poor outcomes and resistance to immunotherapy. We hypothesized that in situ vaccination (ISV) using gene-modified cDC1s engineered to secrete FMS-like tyrosine kinase 3 ligand (FLT3L) would enhance cDC1 function within the TME, promote antitumor immunity, and improve responses to ICB. METHODS: Syngeneic murine models of NSCLC (Kras(G12D)/P53(-/-)/Lkb1(-/-); Kras(G12D)/P53(-/-) ; and Kras(G12D) ) with varying tumor mutational burden, along with the MC38 model, were used to assess the therapeutic efficacy of FLT3L-cDC1 ISV. Flow cytometry and multiplex immunofluorescence were used to evaluate immune mechanisms of response. To assess translational relevance, immune and tertiary lymphoid structure (TLS) signatures were analyzed in The Cancer Genome Atlas (TCGA) NSCLC datasets, with TLS signatures refined using a retrained xCell2 framework incorporating curated TLS and high endothelial venule (HEV) microdissection datasets. RESULTS: FLT3L-cDC1 ISV remodeled the TME across multiple NSCLC models, inducing T lymphocyte infiltration and expanding cytolytic CD8(+) T cells. FLT3L-cDC1 ISV was associated with increased formation of immature TLS with primary follicle-like features within the TME. TCGA analyses revealed that FLT3L expression correlates with activated DC, T cell, and B cell signatures, as well as HEV-enriched TLS-associated programs. Combination with PD-1 blockade further enhanced the antitumor immunity of FLT3L-cDC1 ISV, resulting in robust local and systemic T-cell activation and the expansion of activated CCR7(+)PD-L1(+) cDC1s and stem-like TCF1(+)PD-1(+) CD8(+) progenitors within the TME. In an LKB1-deficient NSCLC model, FLT3L-cDC1 ISV plus PD-1 blockade induced complete and durable regression in 85% of tumors, leading to long-lasting systemic tumor-specific immune memory, consistent with effective tumor vaccination. CONCLUSIONS: FLT3L-cDC1 ISV represents a rational cytokine-enhanced cellular immunotherapy designed to overcome immunosuppression and restore DC function within the TME, thereby promoting tumor-specific adaptive immune responses and enhancing responsiveness to ICB.

Author Info: (1) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (2) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA

Author Info: (1) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (2) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (3) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. VA Greater Los Angeles Healthcare System, Los Angeles, California, USA. (4) Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, Los Angeles, California, USA. (5) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (6) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. VA Greater Los Angeles Healthcare System, Los Angeles, California, USA. (7) Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, Los Angeles, California, USA. (8) Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, Los Angeles, California, USA. (9) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (10) UCLA, Los Angeles, California, USA. (11) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (12) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (13) Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, Los Angeles, California, USA. (14) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. (15) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA. VA Greater Los Angeles Healthcare System, Los Angeles, California, USA. Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, Los Angeles, California, USA. (16) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA rsalehirad@mednet.ucla.edu bliu@mednet.ucla.edu. (17) Department of Medicine, David Geffen School of Medicine, Los Angeles, California, USA rsalehirad@mednet.ucla.edu bliu@mednet.ucla.edu. VA Greater Los Angeles Healthcare System, Los Angeles, California, USA.

B cell-derived type I interferon sustains T cell functionality upon strong TCR stimulation during chronic infection Spotlight 

Graça et al. show that B cells are essential for CD8+ T cell effector response during chronic, but not acute, LCMV infection. In chronic infection, splenic B cells sense viruses via TLR7/8-STING pathways and produce IFN-I, which promotes exhausted T cell differentiation and sustain effector function in the spleen, whereas CD8+ T cells in the LN or responding to low antigen load were B cell independent. Loss of B cells or IFN-I signaling impaired exhausted T cell cytokine production (‘tolerized-like’). IRF1 integrates IFN-I signals to promote T cell expansion. Chronic human HBV showed a similar antigen-load-dependent effect observed in the murine LCMV model.

Contributed by Shishir Pant

Graça et al. show that B cells are essential for CD8+ T cell effector response during chronic, but not acute, LCMV infection. In chronic infection, splenic B cells sense viruses via TLR7/8-STING pathways and produce IFN-I, which promotes exhausted T cell differentiation and sustain effector function in the spleen, whereas CD8+ T cells in the LN or responding to low antigen load were B cell independent. Loss of B cells or IFN-I signaling impaired exhausted T cell cytokine production (‘tolerized-like’). IRF1 integrates IFN-I signals to promote T cell expansion. Chronic human HBV showed a similar antigen-load-dependent effect observed in the murine LCMV model.

Contributed by Shishir Pant

ABSTRACT: B cells are highly abundant lymphocytes and central players in humoral immunity. Although T cells are well known to support humoral responses, how B cells influence T cell responses is less understood. Here, we show that B cells are critical for CD8(+) T cell responses to chronic, but not acute, viral infections. In the absence of B cells, T cells responding to chronic infection exhibited severely impaired effector differentiation. This dependency on B cell help was dictated by high antigen loads and strong T cell receptor (TCR) stimulation. Loss of either B cells or interferon-I (IFN-I) signaling led to severe functional deficits in exhausted T cells, implicating B cells as key producers of IFN-I. The IFN-I-dependent T cell response to strong TCR stimulation is mediated, in part, by the transcription factor IRF1. Therefore, during chronic infection, we uncover an important role for B cell-derived IFN-I in modulating T cell responses to strong TCR stimulation.

Author Info: (1) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (2) Department of Microbi

Author Info: (1) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (2) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (3) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (4) Institute of Molecular Medicine and Experimental Immunology, University Hospital Bonn, University of Bonn, Venusberg-Campus 1, 53127 Bonn, Germany. (5) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia; Institute of Molecular Medicine and Experimental Immunology, University Hospital Bonn, University of Bonn, Venusberg-Campus 1, 53127 Bonn, Germany. (6) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (7) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (8) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (9) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (10) Institute for Immunology, University Hospital Heidelberg, Heidelberg, Germany. (11) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (12) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (13) Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, VIC, Australia; Department of Molecular and Translational Science, Monash University, Clayton, VIC, Australia. (14) Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, VIC, Australia; Department of Molecular and Translational Science, Monash University, Clayton, VIC, Australia. (15) Institute for Immunology, University Hospital Heidelberg, Heidelberg, Germany. (16) Department of Medicine II (Gastroenterology, Hepatology, Endocrinology, and Infectious Diseases), Medical Center, University of Freiburg, Baden-WŸrttemberg, Germany. (17) Department of Medicine II (Gastroenterology, Hepatology, Endocrinology, and Infectious Diseases), Medical Center, University of Freiburg, Baden-WŸrttemberg, Germany. (18) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (19) Institute of Molecular Medicine and Experimental Immunology, University Hospital Bonn, University of Bonn, Venusberg-Campus 1, 53127 Bonn, Germany. (20) Computational Sciences Initiative (CSI), the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (21) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. (22) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. Electronic address: yannick.alexandre@unimelb.edu.au. (23) Department of Microbiology and Immunology, the Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia. Electronic address: daniel.utzschneider@unimelb.edu.au.

IL-2 mutein promotes antigen-specific transplant acceptance in mice through expansion of ST2(+) regulatory T cells Spotlight 

Focused on improving transplantation tolerance, Ganchiku et al. evaluated a long-lived, high-affinity receptor binding IL-2 mutein (mIL-2) in multiple solid organ transplantation models. mIL-2 therapy (with transient co-stimulation blockade) significantly improved long-term allograft survival in an Ag-specific manner, which persisted after cessation of treatment. Allograft survival was accompanied by Treg activation, decreased effector T cell activation, and reduced donor specific antibody levels. mIL-2-mediated allograft survival depended on expansion of highly active and suppressive tissue ST2+ Tregs and was abrogated in Treg-specific ST2 KO graft recipients.

Contributed by Katherine Turner

Focused on improving transplantation tolerance, Ganchiku et al. evaluated a long-lived, high-affinity receptor binding IL-2 mutein (mIL-2) in multiple solid organ transplantation models. mIL-2 therapy (with transient co-stimulation blockade) significantly improved long-term allograft survival in an Ag-specific manner, which persisted after cessation of treatment. Allograft survival was accompanied by Treg activation, decreased effector T cell activation, and reduced donor specific antibody levels. mIL-2-mediated allograft survival depended on expansion of highly active and suppressive tissue ST2+ Tregs and was abrogated in Treg-specific ST2 KO graft recipients.

Contributed by Katherine Turner

ABSTRACT: Although transplantation is the preferred treatment for end-stage organ disease, long-term outcomes are limited by immunosuppressive drug toxicity and immune-mediated injury. Selective in vivo expansion of regulatory T cells (Tregs) using interleukin-2 (IL-2) analogs has emerged as a strategy to induce antigen-specific transplant tolerance with fewer side effects. Herein, we investigate the therapeutic efficacy of an IL-2 mutein molecule (mIL-2) with enhanced receptor specificity and extended half-life in murine models of solid organ transplantation. mIL-2 therapy significantly improves allograft survival in an antigen-specific manner, accompanied by increased Treg activation, decreased effector T cell activation and reduced donor-specific antibody production. Transcriptional profiling reveals expansion of Tregs expressing the suppression of tumorigenicity 2 (ST2) Tregs with heightened activation status and suppressive function. Accordingly, the mIL-2-induced long-term allograft survival is abrogated in Treg-specific ST2 knockout graft recipients, underscoring the critical role of ST2(+) Tregs. These findings identify mIL-2 as an approach to promote long-term transplant tolerance while reducing reliance on conventional immunosuppression.

Author Info: (1) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. Department of Gastroenterological Surg

Author Info: (1) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. Department of Gastroenterological Surgery I, Graduate School of Medicine, Hokkaido University, Hokkaido, Japan. (2) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (3) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (4) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (5) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (6) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (7) Center for Immunology & Inflammatory Diseases, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. Division of Pulmonary and Critical Care Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. (8) Center for Immunology & Inflammatory Diseases, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. Division of Rheumatology, Allergy & Immunology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. (9) Visterra, Inc., Waltham, MA, USA. (10) Visterra, Inc., Waltham, MA, USA. (11) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (12) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (13) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. (14) Center for Transplantation Sciences, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, USA. LRIELLA@mgh.harvard.edu. Nephrology Division, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. LRIELLA@mgh.harvard.edu.

Treg cells promote immunotherapy-induced immune evasion by restraining CD4 T cell control of MHC-I-deficient metastatic pancreatic cancer

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Schmiechen et al. found that variants of PDA that escaped following PD-L1 blockade showed epigenetic silencing of Tap1 expression, which induced loss of IFNγ-inducible MHC-I and reduced tumor cell killing mediated by CD8+ T cell. Escape variants also had an increased capacity for metastasis, which was promoted by Tregs and their suppressive effect on conventional CD4+ T cells. Combination strategies involving restoring IFNγ-inducible MHC-I, targeting Tregs, and enhancing Tconv enabled more effective cancer immunotherapy, suggesting potentially targetable mechanisms to enhance immunotherapy in PDA.

Schmiechen et al. found that variants of PDA that escaped following PD-L1 blockade showed epigenetic silencing of Tap1 expression, which induced loss of IFNγ-inducible MHC-I and reduced tumor cell killing mediated by CD8+ T cell. Escape variants also had an increased capacity for metastasis, which was promoted by Tregs and their suppressive effect on conventional CD4+ T cells. Combination strategies involving restoring IFNγ-inducible MHC-I, targeting Tregs, and enhancing Tconv enabled more effective cancer immunotherapy, suggesting potentially targetable mechanisms to enhance immunotherapy in PDA.

ABSTRACT: Mechanisms driving immunotherapy resistance in pancreatic cancer are poorly defined. We demonstrate that programmed death-ligand 1 immune checkpoint blockade promoted immune evasion by epigenetic Tap1 (transporter associated with antigen processing 1) silencing, increasing selection of metastatic tumor variants with defective interferon-_ (IFN-_)-inducible class I major histocompatibility complex (MHC-I) expression. Unleashing CD4 conventional T cells by regulatory T cell (T(reg) cell) depletion, transfer of tumor-reactive CD4 T cells, or anti-CTLA-4 prevented metastasis. Tumor-specific CD4 T cells adopted a TCF-1(+)SLAMF6(+) progenitor state in lymph nodes and differentiated in tumors. Anti-CTLA-4 increased intratumoral accumulation of CD4 T cells with stemness and tissue residency features, reduced metastasis, and induced gene signatures correlated with improved patient outcomes. MHC-I restoration with anti-CTLA-4 prolonged survival in murine models. In patient tumors, T(reg) cells and CD4 T cells colocalized, and abundance correlated with survival. These findings identify targetable mechanisms of immune evasion and metastasis in immunotherapy-resistant cancer.

Author Info: (1) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapo

Author Info: (1) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (2) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (3) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (4) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (5) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (6) Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA. (7) Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA. (8) Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON, Canada. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada. (9) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (10) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (11) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (12) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (13) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (14) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (15) Institute for Health Informatics, University of Minnesota Medical School, Minneapolis, MN, USA. Clinical Translational Science Institute, University of Minnesota Medical School, Minneapolis, MN, USA. (16) Institute for Health Informatics, University of Minnesota Medical School, Minneapolis, MN, USA. Clinical Translational Science Institute, University of Minnesota Medical School, Minneapolis, MN, USA. (17) Institute for Health Informatics, University of Minnesota Medical School, Minneapolis, MN, USA. Clinical Translational Science Institute, University of Minnesota Medical School, Minneapolis, MN, USA. (18) Caris Life Sciences, Phoenix, AZ, USA. (19) Caris Life Sciences, Phoenix, AZ, USA. (20) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. (21) Institute for Health Informatics, University of Minnesota Medical School, Minneapolis, MN, USA. Clinical Translational Science Institute, University of Minnesota Medical School, Minneapolis, MN, USA. (22) Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON, Canada. Division of Oncology, Department of Statistical Sciences, University of Toronto, Toronto, ON, Canada. Ontario Institute for Cancer Research, Toronto, ON, Canada. Vector Institute, Toronto, ON, Canada. (23) Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA. (24) Department of Microbiology and Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA. Masonic Cancer Center, University of Minnesota Medical School, Minneapolis, MN, USA.

Metabolic determinants of cancer immunotherapy outcomes identified by plasma profiling Spotlight 

Suissa and Fidelle et al. performed targeted metabolomics to 4,336 plasma samples from 1,714 ICI-treated patients across 16 cohorts and trained an ML model that predicted 12‑month PFS, with histidine as a favorable marker and long-chain fatty acids and succinate associated with poor outcome. Histidine supplementation promoted mitochondrial FAO and regulated T cell exhaustion, enhancing ICI-induced antitumor immunity in fibrosarcoma and melanoma models. Histidine-rich diet was associated with favorable PFS in patients without dysbiosis-associated histidine catabolism, and fecal histidine levels inversely correlated with severe irAEs.

Contributed by Shishir Pant

Suissa and Fidelle et al. performed targeted metabolomics to 4,336 plasma samples from 1,714 ICI-treated patients across 16 cohorts and trained an ML model that predicted 12‑month PFS, with histidine as a favorable marker and long-chain fatty acids and succinate associated with poor outcome. Histidine supplementation promoted mitochondrial FAO and regulated T cell exhaustion, enhancing ICI-induced antitumor immunity in fibrosarcoma and melanoma models. Histidine-rich diet was associated with favorable PFS in patients without dysbiosis-associated histidine catabolism, and fecal histidine levels inversely correlated with severe irAEs.

Contributed by Shishir Pant

ABSTRACT: Immune-checkpoint inhibitors benefit a subset of patients with advanced cancer, and the metabolic determinants of response remain unclear. Here, using targeted metabolomics and metagenomics, we profiled 4,336 plasma samples from 1,714 patients across five tumor types and 16 cohorts spanning Europe and North America, longitudinally sampled during five immune-checkpoint inhibitor-based treatment modalities, including fecal microbiota transplantation. A multimodal machine-learning framework integrating 154 metabolites with clinical variables identified five metabolites, age, body mass index and renal function as predictors of 12-month progression-free survival. The model achieved areas under the curve of 0.88 in training and 0.73 in validation cohorts of 105 and 30 patients, respectively and generalized across seven external cohorts. Histidine was a favorable prognostic feature of survival, whereas long-chain fatty acids and succinate were negatively associated with outcome. Histidine supplementation enhanced antitumor immunity in mice. Histidine-rich diets improved progression-free survival in patients lacking dysbiotic microbiome signatures associated with histidine catabolism.

Author Info: (1) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (2) UniversitŽ Paris-Saclay,

Author Info: (1) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (2) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (3) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (4) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (5) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (6) Department of Gastroenterology and Hepatology, University of Groningen and University Medical Center Groningen, Groningen, The Netherlands. Department of Medical Oncology, University of Groningen and University Medical Center Groningen, Groningen, The Netherlands. (7) INSERM U1138 - Metabolism, Cancer & Immunity, ƒquipe LabellisŽe par la Ligue Contre le Cancer, Centre de Recherche des Cordeliers, UniversitŽ Paris CitŽ, Sorbonne UniversitŽ, Paris, France. UniversitŽ Paris-Saclay, INSERM US23 AMMICa, Metabolomic Platform, Gustave Roussy, Villejuif, France. (8) INSERM U1138 - Metabolism, Cancer & Immunity, ƒquipe LabellisŽe par la Ligue Contre le Cancer, Centre de Recherche des Cordeliers, UniversitŽ Paris CitŽ, Sorbonne UniversitŽ, Paris, France. UniversitŽ Paris-Saclay, INSERM US23 AMMICa, Metabolomic Platform, Gustave Roussy, Villejuif, France. (9) Department of Immunology and Genomic Medicine, Center for Cancer Immunotherapy and Immunobiology (CCII), Graduate School of Medicine, Kyoto University, Kyoto, Japan. (10) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (11) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (12) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (13) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. (14) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. Oncoclinicas&Co - Medica Scientia Innovation Research (MEDSIR), Sao Paulo, Brazil. Gonalo Moniz Institute, Fiocruz, Salvador, Brazil. Federal University of Bahia, Salvador, Brazil. (15) Department of Radiation Oncology, Gustave Roussy, UniversitŽ Paris-Saclay, INSERM U1355, RHU LySAIRI, Villejuif, France. (16) Department of Therapeutic Innovation and Early Trials (DITEP), INSERM U981, Gustave Roussy, Villejuif, France. (17) Department of Computational, Cellular and Integrative Biology, University of Trento, Trento, Italy. (18) Department of Computational, Cellular and Integrative Biology, University of Trento, Trento, Italy. (19) Department of Medical Oncology, University of Groningen and University Medical Center Groningen, Groningen, The Netherlands. (20) Verspeeten Family Cancer Centre, London Health Sciences Research Institute, London, Ontario, Canada. Department of Pathology and Laboratory Medicine, Western University, London, Ontario, Canada. Department of Oncology, Division of Experimental Oncology, Schulich School of Medicine & Dentistry, Western University, London, Ontario, Canada. (21) Lawson Health Research Institute, London, Ontario, Canada. Department of Microbiology & Immunology, Western University, London, Ontario, Canada. Department of Medicine, Division of Infectious Diseases, Western University, London, Ontario, Canada. Division of Infectious Diseases, St Joseph's Health Care, London, Ontario, Canada. (22) Verspeeten Family Cancer Centre, London Health Sciences Research Institute, London, Ontario, Canada. Department of Oncology, Western University, London, Ontario, Canada. (23) Department of Twin Research and Genetic Epidemiology, King's College London, London, UK. Department of Dermatology, Mount Vernon Cancer Centre, Northwood, UK. Department of Dermatology, Hemel Hempstead Hospital, West Hertfordshire NHS Trust, Hemel Hempstead, UK. (24) Department of Thoracic Surgery, H™pital-Nord-APHM, Aix-Marseille University, Marseille, France. H™pital Marie Lannelongue, GHPSJ, Le Plessis-Robinson, France. (25) Research Unit Hypertension and Cardiovascular Epidemiology, KU Leuven Department of Cardiovascular Sciences, University of Leuven, Leuven, Belgium. (26) Department of Gastroenterology and Hepatology, University of Groningen and University Medical Center Groningen, Groningen, The Netherlands. (27) Centre de Recherche du Centre Hospitalier de l'UniversitŽ de MontrŽal (CRCHUM), Axe Cancer, Montreal, Quebec, Canada. (28) INSERM U1138 - Metabolism, Cancer & Immunity, ƒquipe LabellisŽe par la Ligue Contre le Cancer, Centre de Recherche des Cordeliers, UniversitŽ Paris CitŽ, Sorbonne UniversitŽ, Paris, France. UniversitŽ Paris-Saclay, INSERM US23 AMMICa, Metabolomic Platform, Gustave Roussy, Villejuif, France. (29) Department of Public Health, Erasmus Medical Centre - University Medical Centre Rotterdam, Rotterdam, The Netherlands. (30) Department of Public Health, Erasmus Medical Centre - University Medical Centre Rotterdam, Rotterdam, The Netherlands. (31) Department of Dermatology, Friedrich-Alexander-UniversitŠt Erlangen-NŸrnberg (FAU), UniversitŠtsklinikum Erlangen, Erlangen, Germany. Bavarian Cancer Research Center (BZKF), Erlangen, Germany. (32) Medical BioSciences, Radboud University Medical Center, Nijmegen, The Netherlands. (33) Centre de Recherche du Centre Hospitalier de l'UniversitŽ de MontrŽal (CRCHUM), Axe Cancer, Montreal, Quebec, Canada. Hemato-Oncology Division, Centre Hospitalier de l'UniversitŽ de MontrŽal (CHUM), Montreal, Quebec, Canada. (34) Dana-Farber Cancer Institute, Boston, MA, USA. Yale School of Medicine, New Haven, CT, USA. (35) Dana-Farber Cancer Institute, Boston, MA, USA. (36) Bavarian Cancer Research Center (BZKF), Erlangen, Germany. Department of Dermatology, University Hospital Regensburg, Regensburg, Germany. (37) Department of Dermatology, Goethe University Frankfurt, University Hospital, Frankfurt am Main, Germany. (38) Department of Translational Medicine and Surgery, Universitˆ Cattolica del Sacro Cuore, Facoltˆ di Medicina e Chirurgia, Rome, Italy. Department of Medical and Surgical Sciences, UOC CEMAD Centro Malattie dell'Apparato Digerente, Medicina Interna e Gastroenterologia, Fondazione Policlinico Universitario Gemelli IRCCS, Rome, Italy. (39) Department of Translational Medicine and Surgery, Universitˆ Cattolica del Sacro Cuore, Facoltˆ di Medicina e Chirurgia, Rome, Italy. Department of Medical and Surgical Sciences, UOC Oncologia Medica, Comprehensive Cancer Center, Fondazione Policlinico Universitario Agostino Gemelli IRCCS, Rome, Italy. (40) Department of Translational Medicine and Surgery, Universitˆ Cattolica del Sacro Cuore, Facoltˆ di Medicina e Chirurgia, Rome, Italy. Department of Medical and Surgical Sciences, UOC Oncologia Medica, Comprehensive Cancer Center, Fondazione Policlinico Universitario Agostino Gemelli IRCCS, Rome, Italy. (41) Unit of Medical Oncology 2, University Hospital of Pisa, Pisa, Italy. Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Pisa, Italy. (42) Dana-Farber Cancer Institute, Boston, MA, USA. Harvard Medical School, Boston, MA, USA. (43) Centre de Recherche du Centre Hospitalier de l'UniversitŽ de MontrŽal (CRCHUM), Axe Cancer, Montreal, Quebec, Canada. Hemato-Oncology Division, Centre Hospitalier de l'UniversitŽ de MontrŽal (CHUM), Montreal, Quebec, Canada. (44) INSERM U1138 - Metabolism, Cancer & Immunity, ƒquipe LabellisŽe par la Ligue Contre le Cancer, Centre de Recherche des Cordeliers, UniversitŽ Paris CitŽ, Sorbonne UniversitŽ, Paris, France. UniversitŽ Paris-Saclay, INSERM US23 AMMICa, Metabolomic Platform, Gustave Roussy, Villejuif, France. Institut du Cancer Paris CARPEM, Department of Biology, H™pital EuropŽen Georges Pompidou, AP-HP, Paris, France. (45) Department of Dermatology, Friedrich-Alexander-UniversitŠt Erlangen-NŸrnberg (FAU), UniversitŠtsklinikum Erlangen, Erlangen, Germany. Bavarian Cancer Research Center (BZKF), Erlangen, Germany. Department of Dermatology and Allergy, LMU University Hospital LMU Munich, Munich, Germany. (46) Department of Immunology and Genomic Medicine, Center for Cancer Immunotherapy and Immunobiology (CCII), Graduate School of Medicine, Kyoto University, Kyoto, Japan. Division of Cancer Immune Regulation, Center for Cancer Immunotherapy and Immunobiology (CCII), Graduate School of Medicine, Kyoto University, Kyoto, Japan. (47) Department of Translational Medicine and Surgery, Universitˆ Cattolica del Sacro Cuore, Facoltˆ di Medicina e Chirurgia, Rome, Italy. Department of Medical and Surgical Sciences, UOC CEMAD Centro Malattie dell'Apparato Digerente, Medicina Interna e Gastroenterologia, Fondazione Policlinico Universitario Gemelli IRCCS, Rome, Italy. (48) Centre de Recherche du Centre Hospitalier de l'UniversitŽ de MontrŽal (CRCHUM), Axe Cancer, Montreal, Quebec, Canada. Hemato-Oncology Division, Centre Hospitalier de l'UniversitŽ de MontrŽal (CHUM), Montreal, Quebec, Canada. (49) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. Department of Medical Oncology, Gustave Roussy, Villejuif, France. (50) MICS Laboratory, CentraleSupŽlec, UniversitŽ Paris-Saclay, Gif-sur-Yvette, France. nikos.paragios@centralesupelec.fr. TheraPanacea, Paris, France. nikos.paragios@centralesupelec.fr. (51) UniversitŽ Paris-Saclay, Gustave Roussy, ClinicObiome, Inserm UMR1367, Microbiota and Mucosal Immunity for Cancer Immunotherapy, Villejuif, France. Laurence.zitvogel@gustaveroussy.fr.

Case of complete response to immunotherapy in MMR-deficient prostate cancer associated with NK-like and CD4+CD8+ T cells

Spotlight 

In a patient with advanced prostate cancer, pembrolizumab resulted in CR and, followed by a radical prostatectomy, prevented recurrence out to 18mo. Tumor whole-exome NGS indicated dMMR/MSI-H and an extremely high TMB. Among PBMCs, CD56+ NK-like CD8+ T cells and CD4+CD8+ double-positive (DP) T cells were observed at high frequency relative to healthy donors, expressed cytotoxic/effector gene signatures and TEMRA phenotype, and clonally expanded after ICB. In other trials (prostate and dMMR/MSI-H cancers), CD56+CD8+ and DP T cells were found among TILs in ICB-treated patients, and DP T cell expansion was positively associated with patient response.

Contributed by Alex Najibi

In a patient with advanced prostate cancer, pembrolizumab resulted in CR and, followed by a radical prostatectomy, prevented recurrence out to 18mo. Tumor whole-exome NGS indicated dMMR/MSI-H and an extremely high TMB. Among PBMCs, CD56+ NK-like CD8+ T cells and CD4+CD8+ double-positive (DP) T cells were observed at high frequency relative to healthy donors, expressed cytotoxic/effector gene signatures and TEMRA phenotype, and clonally expanded after ICB. In other trials (prostate and dMMR/MSI-H cancers), CD56+CD8+ and DP T cells were found among TILs in ICB-treated patients, and DP T cell expansion was positively associated with patient response.

Contributed by Alex Najibi

ABSTRACT: Mismatch repair deficiency (dMMR) and microsatellite instability (MSI-H) are rare in prostate cancer, occurring in 2%-4% of cases. These defects result in increased genomic instability and elevated tumor mutational burden (TMB), which can support responses to immune checkpoint inhibitors (ICIs). Here, we report a patient with locally advanced Gleason 5 + 5 = 10 prostatic adenocarcinoma harboring MSH2 and MSH6 genomic deletions with ultrahigh TMB (>250 mutations/megabase) in whom pembrolizumab resulted in a striking complete radiographic, pathologic, and molecular response. Using digital-spatial microscopy, single-cell RNA/T cell receptor (TCR) sequencing, and multiplex cytometry, we identify atypical tumor-infiltrating T cells with natural killer-like phenotypes and CD4(+)CD8(+) (double-positive) lymphocytes. These clonal T cell populations expand preferentially following ICI and adopt terminally differentiated and cytotoxic profiles that may drive clinical response. Similar T cells are also present in diverse cancers and expand exclusively in ICI-responsive patients. These findings inform on the cellular mechanisms by which immunotherapies may mediate profound responses in patients with dMMR solid tumors.

Author Info: (1) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesot

Author Info: (1) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA; Center for Immunology, University of Minnesota, Minneapolis, MN 55455, USA. (2) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (3) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (4) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (5) Institute for Health Informatics, University of Minnesota, Minneapolis, MN 55455, USA; Clinical Translational Science Institute, University of Minnesota, Minneapolis, MN 55415, USA. (6) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (7) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (8) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. (9) Department of Pharmacology, University of Minnesota, Minneapolis, MN 55455, USA. (10) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (11) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. (12) Division of Infectious Diseases, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. (13) Genitourinary Oncology Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (14) Genitourinary Oncology Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA. (15) Caris Life Sciences, Phoenix, AZ 85040, USA. (16) Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455, USA. (17) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (18) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (19) Allina Health Cancer Institute, Minneapolis, MN 55407, USA. (20) Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA; Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455, USA. (21) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (22) Institute for Health Informatics, University of Minnesota, Minneapolis, MN 55455, USA; Clinical Translational Science Institute, University of Minnesota, Minneapolis, MN 55415, USA. (23) Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA. (24) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. (25) Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. (26) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. (27) Division of Hematology, Oncology and Transplantation, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: anton401@umn.edu.

The transcription factor Eomes drives a stemness program in CD4(+) T cells that promotes anti-tumor immunity in response to immunotherapy Featured  

Agesta, Ferrand, et al. profiled Th antitumor responses and found that Eomes expression and 4-1BB stimulation resulted in Th subsets essential for antitumor responses. Eomes expression resulted in Th subsets: progenitor subsets that self-renew or differentiate into exhausted Th cells via an intermediate effector exhausted state. These cells expanded, infiltrated tumors, and controlled tumor growth. Further, ICB treatment resulted in expansion of Eomes-expressing Th subsets, which accumulated in the tumor, whereas Eomes deficiency limited infiltration and ICB responses.

Agesta, Ferrand, et al. profiled Th antitumor responses and found that Eomes expression and 4-1BB stimulation resulted in Th subsets essential for antitumor responses. Eomes expression resulted in Th subsets: progenitor subsets that self-renew or differentiate into exhausted Th cells via an intermediate effector exhausted state. These cells expanded, infiltrated tumors, and controlled tumor growth. Further, ICB treatment resulted in expansion of Eomes-expressing Th subsets, which accumulated in the tumor, whereas Eomes deficiency limited infiltration and ICB responses.

ABSTRACT: CD4(+) T helper (Th) cells contribute to tumor immunity, yet the subsets and differentiation programs involved remain unclear. Here, we show that the transcription factor Eomesodermin (Eomes) is essential for Th-mediated anti-tumor immunity. Eomes orchestrated the differentiation and maintenance of an exhausted-like Th cell lineage, transcriptionally and functionally distinct from conventional effector or memory Th subsets. This Eomes-dependent program was enhanced by 4-1BB stimulation and promoted effective Th-cell-mediated tumor control. The progenitor subset of this lineage (pTh) expressed stemness-associated transcription factors, displayed self-renewal capacity, and seeded effector subsets capable of controlling tumor growth. At the transcriptional level, Eomes supported the survival, metabolic fitness, and apoptotic resistance of this lineage. Eomes_ pTh cells exhibited conserved transcriptional features in humans across multiple tumor types. As the most expanded Th cell population upon immune checkpoint inhibitor therapy, targeting these cells has potential to improve current immunotherapies.

Author Info: (1) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (2) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (3) University Toulouse, INSERM, CRCT, Toulous

Author Info: (1) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (2) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (3) University Toulouse, INSERM, CRCT, Toulouse, France. (4) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France; ENS, ƒcole Normale SupŽrieure de Lyon, UniversitŽ Claude Bernard - Lyon I, University Lyon, Lyon, France. (5) University Toulouse, INSERM, CRCT, Toulouse, France. (6) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (7) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (8) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (9) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (10) Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA, USA. (11) University Toulouse, INSERM, CRCT, Toulouse, France. (12) University Toulouse, INSERM, CRCT, Toulouse, France. (13) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (14) University Toulouse, INSERM, CRCT, Toulouse, France. (15) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (16) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (17) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (18) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (19) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. (20) Department of Medicine (Medical Oncology) and Neurology, Yale School of Medicine, New Haven, CT, USA. (21) University Toulouse, INSERM, CRCT, Toulouse, France. (22) University Toulouse, INSERM, CRCT, Toulouse, France. (23) University Toulouse, INSERM, CNRS, Infinity, Toulouse, France. Electronic address: anne.dejean@inserm.fr.

A radiopharmaceutical enhances CAR T cells against radio-sensitive and radio-resistant neuroblastoma by tumor sensitization and TME remodeling Spotlight 

Rodriguez and Edinger et al. showed that pre-treatment with VLA-4-targeted, low-dose radiopharmaceutical therapy (RPT) using [67Cu] Cu-LLP2A enhanced GD2 or B7-H3 CAR T cell efficacy, resulting in significant tumor regression in xenogeneic, preclinical neuroblastoma models. The mechanism of action varied with tumor radiosensitivity. In radiosensitive tumors, RPT was directly tumoricidal and enhanced CAR T cell efficacy via TNF-α, leading to paracrine T cell activation. In radioresistant tumors, RPT remodeled the TIME by decreasing the number of M2-like TAMs and stimulating the formation of enriched cytotoxic CD4+ and CD8+ T cell clusters.

Contributed by Katherine Turner

Rodriguez and Edinger et al. showed that pre-treatment with VLA-4-targeted, low-dose radiopharmaceutical therapy (RPT) using [67Cu] Cu-LLP2A enhanced GD2 or B7-H3 CAR T cell efficacy, resulting in significant tumor regression in xenogeneic, preclinical neuroblastoma models. The mechanism of action varied with tumor radiosensitivity. In radiosensitive tumors, RPT was directly tumoricidal and enhanced CAR T cell efficacy via TNF-α, leading to paracrine T cell activation. In radioresistant tumors, RPT remodeled the TIME by decreasing the number of M2-like TAMs and stimulating the formation of enriched cytotoxic CD4+ and CD8+ T cell clusters.

Contributed by Katherine Turner

ABSTRACT: Chimeric antigen receptor (CAR) T cell therapy has limited efficacy against solid tumors such as neuroblastoma (NB). Key obstacles include extensive tumor burden and the presence of an immunosuppressive tumor microenvironment (TME). We employ targeted radiopharmaceutical therapy (RPT) using [(67)Cu]Cu-LLP2A and show that it potentiated the anti-tumor activity of CAR T cells in radio-sensitive and radio-resistant NB models via distinct mechanisms. In radio-sensitive NB, RPT is directly tumoricidal while also enhancing CAR T cell efficacy through pro-immune pathways, most notably via the TNF-_ pathway, leading to paracrine activation of T cells. In radio-resistant NB, RPT improves CAR T cells by remodeling the myeloid compartment in the TME and increasing the formation of immunological niches of cytotoxic CD8(+) GZMB(+) and CD4(+) GZMB(+) CAR T cells. While neither treatment modality alone can effectively treat NB, the combination of VLA-4-targeted RPT and GD2 or B7-H3 CAR T cells augments anti-tumor efficacy, resulting in marked tumor regression in preclinical NB models.

Author Info: (1) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (2) Department of Radiation Oncology, Univer

Author Info: (1) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (2) Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, PA, USA. (3) Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, PA, USA; Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA; Department of Radiology, University of Pittsburgh, Pittsburgh, PA, USA. (4) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (5) Molecular Imaging Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (6) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (7) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (8) Molecular Imaging Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (9) Molecular Imaging Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (10) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (11) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (12) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (13) Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, PA, USA. (14) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (15) Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. (16) Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA; Department of Radiology, University of Pittsburgh, Pittsburgh, PA, USA. (17) Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA; Department of Radiology, University of Pittsburgh, Pittsburgh, PA, USA. (18) Radiation Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (19) Radiation Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (20) Rutgers Cancer Institute, New Brunswick, NJ, USA; Department of Pediatrics, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, New Brunswick, NJ, USA. (21) Departments of Chemistry and Radiology, University of Missouri, Columbia, MO, USA. (22) Molecular Imaging Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA; Radiation Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. Electronic address: freddy.escorcia@gmail.com. (23) Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, PA, USA; Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA; Department of Biomedical Engineering, University of Pittsburgh, Pittsburgh, PA, USA. Electronic address: patelr20@upmc.edu. (24) Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. Electronic address: hongharosa.nguyen@nih.gov.

Activation of tumor-specific CD8+ T cells prior to radiopharmaceutical therapy improves antitumor response Spotlight 

Shim et al. investigated how the timing of tumor antigen-specific CD8+ T cell activation shaped responses to radiopharmaceutical therapy with ⁹⁰Y-NM600 (RPT) in E.G7-OVA and TRAMP-C1 tumor models. Low-dose RPT led to an increase in CD8+ T cell infiltration, but failed to enrich antigen-specific CD8+ T cells. Ex vivo- or (with vaccination) in vivo-activated, but not naive, OT-I cells given prior to RPT slowed tumor growth and expanded antigen-specific effector memory CD8+ T cells in a type I IFN-dependent, cGAS–STING-independent manner. In the TRAMP-C1 prostate cancer model, an AR-encoding DNA vaccine given prior to RPT enhanced tumor control.

Contributed by Shishir Pant

Shim et al. investigated how the timing of tumor antigen-specific CD8+ T cell activation shaped responses to radiopharmaceutical therapy with ⁹⁰Y-NM600 (RPT) in E.G7-OVA and TRAMP-C1 tumor models. Low-dose RPT led to an increase in CD8+ T cell infiltration, but failed to enrich antigen-specific CD8+ T cells. Ex vivo- or (with vaccination) in vivo-activated, but not naive, OT-I cells given prior to RPT slowed tumor growth and expanded antigen-specific effector memory CD8+ T cells in a type I IFN-dependent, cGAS–STING-independent manner. In the TRAMP-C1 prostate cancer model, an AR-encoding DNA vaccine given prior to RPT enhanced tumor control.

Contributed by Shishir Pant

BACKGROUND: Radiopharmaceutical therapy (RPT) delivers radiation systemically, enabling the treatment of metastatic cancers. Beyond killing tumor cells, RPT can modulate the tumor immune microenvironment. With RPTs and immunotherapies already approved or in development for prostate cancer, many preclinical and clinical studies are evaluating their use in combination. However, due to the radiosensitivity of tumor-infiltrating lymphocytes, further studies are needed to determine the effects of RPT on these cells to better inform the sequence of immunotherapies that activate T cells when given with RPT. METHODS: E.G7-OVA tumor-bearing mice received na•ve or activated OT-I CD8+T cells prior to or following the administration of RPT using (90)Y-NM600. Changes in tumor growth were monitored, and tumor-infiltrating lymphocytes were evaluated for phenotypic and functional markers. The murine prostate tumor model TRAMP-C1 was used to evaluate this approach using tumor antigen-specific vaccination with (90)Y-NM600. RESULTS: Antitumor efficacy was improved if OT-I CD8+T cells were present and activated prior to (90)Y-NM600 administration than if the cells were delivered after RPT. Similarly, in vivo activation of adoptively transferred OT-I CD8+T cells, using ovalbumin (OVA)-specific vaccination, prior to RPT slowed tumor growth and increased the frequency of tumor-infiltrating OVA(257-264)-specific CD8+T cells with effector memory phenotype and effector molecule production. Blockade of type I interferon, but not the upstream inhibition of stimulator of interferon genes, abrogated tumor growth delay resulting from the combination treatment. Tumor antigen-specific vaccination prior to (90)Y-NM600 administration similarly improved antitumor outcomes in the TRAMP-C1 tumor model. CONCLUSIONS: Our study suggests that tumor-specific CD8+T cells need to be present and activated prior to RPT to enhance antitumor outcomes. This study highlights the importance of considering the effects of RPT on tumor-infiltrating CD8+T cells when combining other T-cell activating therapies with RPT, as they may similarly display sequence-dependent antitumor outcomes.

Author Info: (1) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (2) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (3) University of Wisconsin

Author Info: (1) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (2) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (3) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (4) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (5) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (6) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (7) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (8) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA. (9) University of Wisconsin Carbone Cancer Center, Madison, Wisconsin, USA dm3@medicine.wisc.edu.

CD4+ T cells impair tumor growth through IL-3 and TNF-dependent vascular damage

Spotlight 

Lian and Nie et al. showed that LCMV gp66-specific CD4+ T cells inhibited tumor growth in an MC38-GP model in an antigen-specific manner, independent of direct lymphoid cell-mediated cytotoxicity. CD4+ T cells initiated antigen-dependent perivascular, myeloid cell-dense structures in the TIME, reprogrammed myeloid transcriptomes, and leveraged recruited myeloid cells to control tumor growth. Single-cell and spatial transcriptomics showed that CD4+ T cell-derived IL-3 programmed macrophages to secrete tumor necrosis factor, which damaged intratumoral vasculature, compromised blood supply, and induced localized tumor cell death and regression.

Contributed by Shishir Pant

Lian and Nie et al. showed that LCMV gp66-specific CD4+ T cells inhibited tumor growth in an MC38-GP model in an antigen-specific manner, independent of direct lymphoid cell-mediated cytotoxicity. CD4+ T cells initiated antigen-dependent perivascular, myeloid cell-dense structures in the TIME, reprogrammed myeloid transcriptomes, and leveraged recruited myeloid cells to control tumor growth. Single-cell and spatial transcriptomics showed that CD4+ T cell-derived IL-3 programmed macrophages to secrete tumor necrosis factor, which damaged intratumoral vasculature, compromised blood supply, and induced localized tumor cell death and regression.

Contributed by Shishir Pant

ABSTRACT: Most cancer immunotherapy strategies are focused on direct tumor killing by immune cells, especially T lymphocytes. Clinical and conceptual limitations of these approaches create a need for additional strategies. We identified a tumor stroma-targeting mechanism in which tumor antigen-specific CD4(+) T cells inhibit tumor growth through myeloid cell and tumor necrosis factor (TNF)-dependent vascular damage. Multiplex immunofluorescence and single-cell and tissue transcriptomics showed that CD4(+) T cells trigger the formation of perivascular myeloid cell clusters containing "classically activated" macrophages that produce TNF in response to T cell-derived interleukin-3. TNF causes intratumoral endothelial damage and blood supply disruption, which are associated with localized tumor cell death. Thus, intratumoral antigen-triggered T cell activation can mediate antitumor effects without direct recognition of living tumor cells, thereby avoiding many of the inhibitory mechanisms that limit anti-tumor immunity.

Author Info: (1) Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. (2) La

Author Info: (1) Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. (2) Laboratory of Immune Cell Biology and Laboratory of Integrative Cancer Immunology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (3) Single Cell Analysis Facility, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (4) Laboratory of Immune Cell Biology and Laboratory of Integrative Cancer Immunology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (5) Molecular Histopathology Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, USA. (6) Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. (7) Laboratory of Immune Cell Biology and Laboratory of Integrative Cancer Immunology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (8) Laboratory of Immune Cell Biology and Laboratory of Integrative Cancer Immunology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (9) Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. (10) Laboratory of Immune Cell Biology and Laboratory of Integrative Cancer Immunology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (11) Molecular Histopathology Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, USA. (12) Single Cell Analysis Facility, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. (13) Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. (14) Laboratory of Immune Cell Biology and Laboratory of Integrative Cancer Immunology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

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