Journal Articles

Oncogenic Kras targeting with MRTX1133 or Daraxonrasib specifically synergize with anti-CTLA4 to promote anti-tumor immunity in pancreatic cancer

ABSTRACT: Lack of sustained response to oncogenic Kras (Kras*) inhibition in pancreatic ductal adenocarcinoma (PDAC) underscores the need to identify effective combination therapies. Here, we demonstrate that Kras* targeting using MRTX1133 or Daraxonrasib recruits diverse T-cell infiltrates, including regulatory (Tregs), effector and exhausted T cells into the PDAC microenvironment. Kras* inhibition induces T-cell influx and offers a therapeutic window to specifically prime PDAC to anti-CTLA4 immune checkpoint blockade efficacy, in contrast to anti-PD1, anti-Tim3, anti-Lag3, anti-Vista, and anti-4-1BB agonist combination therapy. Mechanistically, anti-CTLA4 combination therapy transcriptionally reprograms effector Tregs to a naive phenotype, reverses CD8+ T-cell exhaustion, and promotes recruitment of functional tertiary lymphoid structures to mediate anti-tumor immunity. Single-cell ATAC sequencing reveals that Treg reprogramming by anti-CTLA4 is epigenetically regulated by downregulation of AP-1 family transcription factors in the IL-35 promoter region. This study reveals an actionable vulnerability in the adaptive immune response in Kras* targeted PDAC with immediate clinical implications.

Author Info: (1) Department of Cancer Biology, University of Texas MD Anderson Cancer Center, Houston, TX, USA (2) 3P-Medicine Laboratory, Medical University of Gdansk, 80-210 Gdansk, Poland (3

Author Info: (1) Department of Cancer Biology, University of Texas MD Anderson Cancer Center, Houston, TX, USA (2) 3P-Medicine Laboratory, Medical University of Gdansk, 80-210 Gdansk, Poland (3) Departments of Anatomical Pathology and Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, TX, USA (4) Department of Investigative Cancer Therapeutics, University of Texas MD Anderson Cancer Center, Houston, TX, USA (5) Institute for Applied Science and TRACTION platform, University of Texas MD Anderson Cancer Center, Houston, TX, USA (6) Department of Bioengineering, Rice University, Houston, TX, USA (7) Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA (8) Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA

The Dendritic Cell-based Vaccine PROTEXI leverages Antiviral CD4 T cell Memory to boost anti-tumor immune responses in mice

The efficacy of dendritic cell (DC) cancer vaccines is linked to poor immunogenicity of tumor-associated antigens and failure to elicit robust MHC class II-restricted CD4_ T-cell responses. Here, we introduce PROTEXI, a DC vaccine platform that optimizes tumor immunity by co-presenting tumor-specific CD8_ T-cell epitopes alongside CD4_ T helper epitopes from the SARS-CoV-2 Spike protein, leveraging widespread anti-viral immunity. In preclinical mouse models of melanoma and breast cancer, PROTEXI significantly reduces tumor growth and improves survival by promoting robust T cell infiltration into immune-cold tumors, increasing cytotoxic T cell responses via epitope spreading, and activating genes linked to optimal DC, NK cell, and T cell function. Furthermore, PROTEXI elicits superior responses when combined with other immunotherapy agents in models of therapy-resistant tumors. Finally, in a humanized mouse model of melanoma, PROTEXI vaccine co-presenting CD4_ T-specific Spike epitopes with CD8_ T cell-restricted PRAME and MAGE-A3 antigens, significantly reduces tumor burden. Thus, these data underscore the potential of harnessing pre-existing viral-specific immunity to enhance the efficacy of DC vaccines in immune-cold tumors.

Author Info: (1) The Angie Fowler Adolescent & Young Adult Cancer Institute, University Hospitals Rainbow Babies & Children's Hospital, Cleveland, OH, USA. Department of Pediatric Hematology &

Author Info: (1) The Angie Fowler Adolescent & Young Adult Cancer Institute, University Hospitals Rainbow Babies & Children's Hospital, Cleveland, OH, USA. Department of Pediatric Hematology & Oncology, University Hospitals Cleveland Medical Center, Cleveland, OH, USA. (2) Department of Pediatrics, Case Western Reserve University, Cleveland, OH, USA. (3) Celloram Inc., 11000 Cedar Avenue STE 100F #23, Cleveland, OH, USA. (4) Department of Pediatrics, Case Western Reserve University, Cleveland, OH, USA. (5) Celloram Inc., 11000 Cedar Avenue STE 100F #23, Cleveland, OH, USA. (6) Celloram Inc., 11000 Cedar Avenue STE 100F #23, Cleveland, OH, USA. (7) MedPacto Inc., 92, Myeongdal-ro, Seocho-gu, Seoul, Republic of Korea. (8) The Angie Fowler Adolescent & Young Adult Cancer Institute, University Hospitals Rainbow Babies & Children's Hospital, Cleveland, OH, USA. John.Letterio@UHHospitals.org. Department of Pediatric Hematology & Oncology, University Hospitals Cleveland Medical Center, Cleveland, OH, USA. John.Letterio@UHHospitals.org. Department of Pediatrics, Case Western Reserve University, Cleveland, OH, USA. John.Letterio@UHHospitals.org. The Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA. John.Letterio@UHHospitals.org. (9) The Angie Fowler Adolescent & Young Adult Cancer Institute, University Hospitals Rainbow Babies & Children's Hospital, Cleveland, OH, USA. SeunghwanLim@Celloram.com. Celloram Inc., 11000 Cedar Avenue STE 100F #23, Cleveland, OH, USA. SeunghwanLim@Celloram.com.

Tumor-induced dendritic cell deregulation perturbs T cell proliferation and predicts clinical outcome in acute lymphoblastic leukemia

Perturbations in dendritic cells (DCs) in B/T cell acute lymphoblastic leukemia (ALL), their cause(s) and consequence(s) on antileukemia immunity, and patient outcomes remain poorly explored. We find that maturation of DC1-6 subsets is disrupted in children and adults with ALL. Conventional DC1 and DC2 subpopulations are reduced at the expense of the progenitors and the DC4 fractions in ALL. The potential to mature, present antigens, and produce cytokines for initiating T cell surveillance appears to be impaired in every ALL DC subset. Such DC subsets are accordingly unable to induce T cell proliferation compared to DCs from healthy donors. MYC overexpression in ALL cells disrupts DC homeostasis and reduces the ability of DCs to induce T cell proliferation. Predominance of cells with transcriptional signatures typical of "stimulated DCs" predicts favorable clinical outcomes in B-ALL, while it is associated with unfavorable outcomes in T-ALL. Phenotyping DC subsets at ALL diagnosis could thus be valuable in informing treatment outcomes.

Author Info: (1) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (2) Department of Systems Biology, Beckman Research Institute of City of Hop

Author Info: (1) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (2) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (3) The Human Immune Monitoring Center (HIMC), Institute for Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, CA 94305, USA. (4) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (5) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (6) Department of Molecular and Cellular Biology, City of Hope National Medical Center, Duarte, CA 91010, USA. (7) The Hematopoietic Tissue Biorepository/Research Pathology Shared Resources, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA. (8) Department of Pediatrics (Hematology and Oncology), Stanford University School of Medicine, Stanford, CA 94305, USA. (9) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (10) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (11) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. (12) The Hematopoietic Tissue Biorepository/Research Pathology Shared Resources, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA. (13) The Hematopoietic Tissue Biorepository/Research Pathology Shared Resources, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA; The Department of Hematological Malignancies Translational Science, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA. (14) Department of Pediatrics (Hematology and Oncology), Stanford University School of Medicine, Stanford, CA 94305, USA. (15) Department of Pediatrics (Hematology and Oncology), Stanford University School of Medicine, Stanford, CA 94305, USA. (16) Department of Basic Science, Division of Cancer Sciences, Loma Linda University School of Medicine, Loma Linda, CA 92350, USA. (17) Department of Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA. (18) Division of Oncology and Center for Childhood Cancer Research, Department of Pediatrics, Children's Hospital of Philadelphia, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Department of Pediatrics and Abramson Cancer Center, University of Pennsylvania School of Medicine, Philadelphia, PA, USA. (19) Department of Laboratory Medicine, University of California, San Francisco, San Francisco, CA 94143, USA. (20) Department of Clinical Population and Public Health Sciences, University of Southern California, Los Angeles, CA 91016, USA. (21) Department of Pediatrics, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA. (22) Schneider Children's Medical Center, Tel Aviv University, Felsenstein Research Institute, Tel Aviv, Israel. (23) Department of Molecular and Cellular Biology, City of Hope National Medical Center, Duarte, CA 91010, USA. (24) The Human Immune Monitoring Center (HIMC), Institute for Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, CA 94305, USA. (25) Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA; Department of Pediatrics, Beckman Research Institute of City of Hope, Duarte, CA 91010, USA; Center for RNA Biology and Therapeutics, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA. Electronic address: sswaminathan@coh.org.

CCR7+ activated dendritic cells are essential for spontaneous and immunotherapy-driven anti-tumor immunity

Single-cell transcriptomics identifies a convergent activation state of conventional dendritic cells (cDCs) shared by type 1 and type 2 cDCs (cDC1s and cDC2s). These activated DCs (actDCs) are characterized by co-expression of T cell-stimulating and inhibitory molecules. Here, we examined the functional contribution of actDCs to anti-tumor immunity by developing mouse models that leverage CCR7 expression to conditionally label or ablate actDCs. The capacity of cDCs to stimulate tumor-specific cytotoxic T lymphocytes (CTLs) was restricted to the actDC state. cDC1- and cDC2-derived actDCs supported CTL priming through cross-presentation and cross-dressing, respectively, with the latter occurring in a cancer type-dependent manner. actDCs were required for the activation of naive CTLs in tumor-draining lymph nodes and for sustaining effector CTL function within tumors. Consequently, ablation of actDCs impaired spontaneous tumor control and responses to immune checkpoint blockade or adoptive T cell therapy. Thus, the actDC state emerges as a critical determinant of cDC-mediated anti-tumor immunity.

Author Info: (1) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (2) Cancer Inflammation and Immunity, Cancer Research U

Author Info: (1) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (2) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (3) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (4) Computational Biology Support, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (5) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (6) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (7) Flow Cytometry, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (8) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (9) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK; Dermatology Centre, Northern Care Alliance NHS Foundation Trust & Division of Musculoskeletal and Dermatological Sciences, Manchester NIHR Biomedical Research Centre, Manchester Academic Health Science Centre, The University of Manchester, Manchester, UK. (10) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (11) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (12) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (13) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (14) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (15) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (16) Computational Biology Support, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (17) Centre d'Etude des Pathologies Respiratoires, INSERM, UMR 1100, UniversitŽ de Tours, Tours, France. (18) Centre d'Immunologie de Marseille-Luminy, INSERM, CNRS, Aix Marseille UniversitŽ, Marseille, France; Centre d'ImmunophŽnomique, INSERM, CNRS, PHENOMIN, Celphedia, Aix Marseille UniversitŽ, Marseille, France. (19) Lydia Becker Institute of Immunology and Inflammation, The University of Manchester, Manchester, UK. (20) Genome Editing and Mouse Models, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK. (21) Cancer Inflammation and Immunity, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, UK; Lydia Becker Institute of Immunology and Inflammation, The University of Manchester, Manchester, UK. Electronic address: santiago.zelenay@cruk.manchester.ac.uk.

Spatial biology reveals altered macrophage states in immunosuppressed non-melanoma skin cancer

Immunosuppressed patients with non-melanoma skin cancer experience worse clinical outcomes, yet the tumor immune microenvironment associated with systemic immunosuppression remains incompletely defined. Using integrated single-cell, spatial transcriptomic, multiplex immunofluorescence, and spatial epigenomic profiling across immunocompetent and immunosuppressed tumors, we found that overall immune-cell composition was largely preserved despite differences in immune-cell distribution, spatial organization, and T cell clonality. Immunosuppressed tumors demonstrated reduced intratumoral macrophage densities, decreased T cell clonal diversity, altered antigen-presenting cell and T cell spatial interactions, and distinct fibroblast- and macrophage-associated spatial niches. Multi-cohort validation across complementary spatial and single-cell platforms identified consistent alterations in innate-adaptive immune organization in immunosuppressed tumors. Together, these findings define spatial and functional remodeling of the tumor immune microenvironment under systemic immunosuppression and provide a framework for future therapeutic investigation in high-risk patients.

Author Info: (1) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (2) MD Anderson Epigenomics Therapy Initiative, Department of Genomic

Author Info: (1) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (2) MD Anderson Epigenomics Therapy Initiative, Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (3) Department of Bioinformatics and Computational Biology, Division of Discovery Science, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (4) MD Anderson Epigenomics Therapy Initiative, Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (5) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Otolaryngology-Head and Neck Surgery, Gleiberman Head and Neck Cancer Center, Moores Cancer Center, University of California, San Diego, La Jolla, CA, USA. (6) MD Anderson Epigenomics Therapy Initiative, Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (7) AtlasXomics, Pierce Laboratory, 290 Congress Ave, New Haven, CT, USA. (8) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (9) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. Electronic address: fonetto@mdanderson.org. (10) Department of Anatomical Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. Electronic address: pnagarajan@mdanderson.org. (11) Department of Cancer Sciences, Cleveland Clinic Research, Cleveland Clinic, Cleveland, OH, USA. (12) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (13) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (14) Department of Radiation Oncology, Cleveland Clinic, Cleveland, OH, USA. (15) Department of Radiation Oncology, Cleveland Clinic, Cleveland, OH, USA. (16) Department of Otolaryngology, Head and Neck Surgery, Rabin Medical Center, Petah Tikva, Israel. (17) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (18) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (19) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (20) MD Anderson Epigenomics Therapy Initiative, Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (21) AtlasXomics, Pierce Laboratory, 290 Congress Ave, New Haven, CT, USA. (22) Department of Biology and Biochemistry, University of Houston Sequencing Core, University of Houston, Houston, TX, USA; Department of Molecular and Cellular Biology, Human Genome Sequencing Center, Baylor College of Medicine, Houston, TX , USA. (23) Department of Biology and Biochemistry, University of Houston Sequencing Core, University of Houston, Houston, TX, USA; Department of Molecular and Cellular Biology, Human Genome Sequencing Center, Baylor College of Medicine, Houston, TX , USA. (24) Department of Dermatology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. Electronic address: mrmigden@mdanderson.org. (25) Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (26) Division of Cancer Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (27) Department of Pathology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA. (28) Department of Cancer Sciences, Cleveland Clinic Research, Cleveland Clinic, Cleveland, OH, USA. Electronic address: mcgraid@ccf.org. (29) Department of Bioinformatics and Computational Biology, Division of Discovery Science, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (30) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (31) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (32) MD Anderson Epigenomics Therapy Initiative, Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. Electronic address: krai@mdanderson.org. (33) Department of Head and Neck Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; UTHealth Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Cancer Neuroscience Program, UT MD Anderson Cancer Center, Houston, TX, USA. Electronic address: mamit@mdanderson.org.

Dendritic cell circadian clocks shape memory CD8+ T cell differentiation

Circadian rhythms regulate diverse immune processes, yet how they influence memory CD8(+) T cell differentiation remains unclear. Here, we show that the time of day of antigen encounter shapes CD8(+) T cell fate and antiviral immunity. Immunization during the active phase promotes the generation of progenitor-like memory CD8(+) T cells and enhances T cell-mediated protection upon viral challenge. Mechanistically, dendritic cell-intrinsic circadian clocks regulate expression of the costimulatory ligand CD70, thereby directing T cell differentiation. These findings uncover a dendritic cell-mediated circadian mechanism that governs memory T cell fate decisions and suggest that aligning immune priming with circadian time may be leveraged to optimize T cell immunity.

Author Info: (1) Department of Immunology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (2) Department of Immunology, Leiden University Medical Center, Alb

Author Info: (1) Department of Immunology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (2) Department of Immunology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (3) Department of Immunology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (4) Department of Immunology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (5) Leiden University Center for Infectious Diseases, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (6) Leiden University Center for Infectious Diseases, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (7) Leiden University Center for Infectious Diseases, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands. (8) Department of Cell and Chemical Biology, Leiden University Medical Center, Einthovenweg 20, 2333 ZC Leiden, The Netherlands. (9) Department of Immunology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.

First-in-human testing of a mutant KRAS vaccine for pancreatic cancer interception in high-risk cohorts

Pancreatic ductal adenocarcinoma (PDAC) arises from precursor lesions over a decade-plus, offering a window for interception in high-risk individuals, but current surveillance detects a minority of precursors. Mutant KRAS (mKRAS) is present in most PDACs and their precursors, making it an appealing target for immune-based interception. We conducted a phase I, first-in-human study of a peptide vaccine targeting six common KRAS mutations (mKRAS-VAX) in 20 individuals with hereditary PDAC predisposition and a radiographic pancreatic abnormality (NCT05013216) to assess safety, immunogenicity, and T cell persistence. Adverse events were grade 1-2. Vaccination elicited a significant mKRAS-specific T cell response in 18/20 participants (90%). Longitudinal TCR sequencing demonstrated persistence of vaccine-induced mKRAS-specific clonotypes for up to 2 years. Over a median follow-up of 16.5 months, no participants developed PDAC. These findings demonstrate that mKRAS-VAX is safe and generates durable T cell responses, which support the advancement of mKRAS-targeted vaccination for PDAC interception.

Author Info: (1) The University of Texas MD Anderson Cancer Center Houston, TX United States. ROR: https://ror.org/04twxam07 (2) Johns Hopkins Medicine Baltimore United States. ROR: https://ror

Author Info: (1) The University of Texas MD Anderson Cancer Center Houston, TX United States. ROR: https://ror.org/04twxam07 (2) Johns Hopkins Medicine Baltimore United States. ROR: https://ror.org/037zgn354 (3) Johns Hopkins Medicine Baltimore, Maryland United States. ROR: https://ror.org/037zgn354 (4) Johns Hopkins Medicine Baltimore, Maryland United States. ROR: https://ror.org/037zgn354 (5) Johns Hopkins Medicine Baltimore, MD United States. ROR: https://ror.org/037zgn354 (6) Johns Hopkins University United States. ROR: https://ror.org/00za53h95 (7) Johns Hopkins Medicine Baltimore, MD United States. ROR: https://ror.org/037zgn354 (8) Johns Hopkins School of Medicine Baltimore, MD United States. (9) Icahn School of Medicine at Mount Sinai New York United States. ROR: https://ror.org/04a9tmd77 (10) Johns Hopkins Medicine Baltimore, MD United States. ROR: https://ror.org/037zgn354 (11) Johns Hopkins Medicine Baltimore United States. ROR: https://ror.org/037zgn354 (12) The Johns Hopkins Medical Institutions Baltimore, MD United States. (13) Johns Hopkins University Baltimore, MD United States. ROR: https://ror.org/00za53h95 (14) Johns Hopkins University Baltimore, MD United States. ROR: https://ror.org/00za53h95 (15) Johns Hopkins Medicine Baltimore United States. ROR: https://ror.org/037zgn354 (16) Johns Hopkins University Baltimore, MD United States. ROR: https://ror.org/00za53h95 (17) Johns Hopkins University Baltimore, MD United States. ROR: https://ror.org/00za53h95 (18) Sidney Kimmel Comprehensive Cancer Center Baltimore, Maryland United States. ROR: https://ror.org/05m5b8x20 (19) Johns Hopkins Medicine Baltimore, MD United States. ROR: https://ror.org/037zgn354 (20) Johns Hopkins Medicine Baltimore, MD United States. ROR: https://ror.org/037zgn354 (21) Sidney Kimmel Comprehensive Cancer Center Baltimore, MD United States. ROR: https://ror.org/05m5b8x20 (22) Johns Hopkins University Baltimore, MD United States. ROR: https://ror.org/00za53h95 (23) Johns Hopkins Medicine Baltimore, MD United States. ROR: https://ror.org/037zgn354 (24) Johns Hopkins Medicine Baltimore United States. ROR: https://ror.org/037zgn354 (25) Johns Hopkins University Baltimore, Maryland United States. ROR: https://ror.org/00za53h95 (26) Johns Hopkins University Baltimore, MD United States. ROR: https://ror.org/00za53h95 (27) Johns Hopkins University Baltimore, MD United States. ROR: https://ror.org/00za53h95 (28) Johns Hopkins Medicine Baltimore, Maryland United States. ROR: https://ror.org/037zgn354

CD4+ T cells play distinct roles during the priming versus effector phases of immune checkpoint therapy-dependent tumor elimination by CD8+ T cells Featured  

Ameh et al. studied the role of CD4+ T cells in mouse models during ICB treatment. CD4+ T cells were crucial for priming and effector phases to induce tumor rejection. During the effector phase, CD4+ T cells helped reinvigorate tumor-specific CD8+ T cells. CD40/CD40L signaling was vital for CD4+ T cell function during priming, while IL-2 and IFNγ were necessary for antitumor CD8+ T cell responses during the effector phase.

Ameh et al. studied the role of CD4+ T cells in mouse models during ICB treatment. CD4+ T cells were crucial for priming and effector phases to induce tumor rejection. During the effector phase, CD4+ T cells helped reinvigorate tumor-specific CD8+ T cells. CD40/CD40L signaling was vital for CD4+ T cell function during priming, while IL-2 and IFNγ were necessary for antitumor CD8+ T cell responses during the effector phase.

ABSTRACT: It is well established that CD4⁺ T cells play a critical role in facilitating immune checkpoint therapy (ICT). Although CD4+ T-cell function in lymph nodes during CD8⁺ T-cell priming has been well investigated, their requirement at the effector phase in the tumor is only now beginning to be appreciated. Herein, we used our major histocompatibility complex class II-negative (MHC-II-) sarcoma models to confirm that CD4⁺ T cells are essential not only during T-cell priming, but also to maintain T-cell effector function within the tumor. Depleting CD4⁺ T cells at the effector phase, after CD8+ T-cell priming had occurred, abolished ICT-induced tumor rejection despite the detection of tumor-specific CD8⁺ T cells and their intratumoral accumulation. CD4⁺ T cells were required for functional reinvigoration of CD8⁺ tumor-infiltrating lymphocytes (TIL) by ICT, leading to enhanced cytokine production, expression of cytotoxicity, and reduced exhaustion-without affecting CD8+ T-cell proliferation. Mechanistically, CD4⁺ T-cell function at the effector phase did not require CD40/CD40L signaling, which is necessary for efficient priming, but rather depended on IL-2 and IFNγ. Using a TCR-mimic monoclonal antibody (1G10) specific for the dominant neoantigen:I-Aᵇ complex on antigen-presenting cells formed during T3 sarcoma challenge, we further showed that ongoing MHC-II neoantigen presentation was necessary to sustain CD4⁺ T-cell help after priming. These findings reveal temporally distinct requirements for CD4⁺ T-cell help and establish a need for continuous CD4⁺/CD8⁺ T-cell cooperation as a prerequisite for anti-PD-1/anti-CTLA-4 ICT efficacy against MHC-II- tumors.

Author Info: (1) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (2) Washington University in St. Louis St. Louis, MO United States. ROR: https://

Author Info: (1) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (2) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (3) Washington University in St Louis St Louis United States. (4) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (5) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (6) Washington University in St. Louis United States. ROR: https://ror.org/01yc7t268 (7) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (8) Washington University in St. Louis St. Louis, Missouri United States. ROR: https://ror.org/01yc7t268 (9) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (10) Washington University in St. Louis Saint Louis, MO United States. ROR: https://ror.org/01yc7t268 (11) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (12) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268 (13) Washington University in St. Louis St. Louis, Missouri United States. ROR: https://ror.org/01yc7t268 (14) Washington University in St. Louis St. Louis, Missouri United States. ROR: https://ror.org/01yc7t268 (15) Washington University in St. Louis St. Louis, Missouri United States. ROR: https://ror.org/01yc7t268 (16) Washington University in St. Louis St. Louis, MO United States. ROR: https://ror.org/01yc7t268

Immunologic determinants of infusion products and the tumor microenvironment govern response to TIL therapy in advanced melanoma Spotlight 

Karapetyan et al. profiled TIL infusion products and matched TIMEs from patients with metastatic melanoma treated on TIL-based trials with or without ICB or BRAFi, and found that responders had infusion products enriched for stem-like memory and LAG3+ CD8+ TILs with enhanced peripheral persistence. Multiplex IF and spatial transcriptomics identified TLSs and immune transcriptomic features, such as antigen presentation, IFN signaling, B cell activation, and chemokine pathways in TIL responders. Prior ICB exposure reduced CD8+ stem-like TILs, co-stimulatory receptor expression, and TCR diversity, consistent with impaired TIL fitness.

Contributed by Shishir Pant

Karapetyan et al. profiled TIL infusion products and matched TIMEs from patients with metastatic melanoma treated on TIL-based trials with or without ICB or BRAFi, and found that responders had infusion products enriched for stem-like memory and LAG3+ CD8+ TILs with enhanced peripheral persistence. Multiplex IF and spatial transcriptomics identified TLSs and immune transcriptomic features, such as antigen presentation, IFN signaling, B cell activation, and chemokine pathways in TIL responders. Prior ICB exposure reduced CD8+ stem-like TILs, co-stimulatory receptor expression, and TCR diversity, consistent with impaired TIL fitness.

Contributed by Shishir Pant

BACKGROUND: The immunologic features of tumor-infiltrating lymphocyte (TIL) infusion products and their interactions with the tumor microenvironment that govern clinical responses in metastatic melanoma remain incompletely characterized. METHODS: We performed integrated immunophenotypic and spatial transcriptomic profiling of TIL infusion products and matched tumor microenvironments from patients treated on early-phase clinical trials. RESULTS: The durable objective response rate was 36%, with a median progression-free survival of 8 months among patients treated with TIL therapy with or without additional therapies. Responders exhibited infusion products enriched for CD8(+) T cells, stem-like memory subsets, and LAG-3-expressing CD8(+) T cells, together with enhanced peripheral TIL persistence. The abundance of infused LAG-3+ TILs correlated with tumor reactivity and prolonged progression-free survival. Prior immune checkpoint inhibitor exposure was associated with reduced CD8(+) stem cell memory T cell frequency and diminished co-stimulatory receptor expression, suggesting impaired TIL fitness. Tumors enriched for tertiary lymphoid structures and spatial immune programs characterized by antigen presentation, interferon signaling, and B cell activation were independently associated with clinical benefit. CONCLUSIONS: These findings define an integrated framework linking TIL composition and the tumor microenvironment to therapeutic response and identify potential biomarkers and biological features that may guide patient selection and optimization of TIL therapy. FUNDING: This work was funded by Iovance Biotherapeutics, the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation, the Melanoma Research Alliance, the Donald A. Adam Melanoma & Skin Cancer Center of Excellence, American Cancer Society-Leo and Anne Albert Charitable Foundation Research Scholar Grant, and Melanoma SPORE (P50CA168536).

Author Info: (1) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School

Author Info: (1) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. Electronic address: lilit.karapetyan@moffitt.org. (2) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA. (3) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (4) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA. (5) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (6) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (7) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (8) Department of Molecular Biosciences, University of South Florida, Tampa, FL 33612, USA. (9) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (10) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (11) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (12) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (13) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (14) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA. (15) Department of Sarcoma, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (16) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA. (17) Department of Sarcoma, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA. (18) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA. (19) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA; Department of Anatomic Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (20) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA; Department of Anatomic Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (21) Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (22) Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (23) Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (24) Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (25) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA. (26) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (27) Department of Sarcoma, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA. (28) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA. (29) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (30) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA. (31) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (32) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. (33) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Oncologic Sciences, University of South Florida School of Medicine, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. Electronic address: amod.sarnaik@moffitt.org. (34) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. Electronic address: matthew.beatty@moffitt.org. (35) Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Immuno-Oncology Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA; Department of Translational Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. Electronic address: shari.pilon-thomas@moffitt.org.

Tumor-specific antibodies elicited by engineered bacteria promote bladder cancer immunotherapy in preclinical mouse models Spotlight 

Rouanne et al. engineered probiotic E.coli (EcN) using a synchronized lysis integrated circuit to release human CXCL13 at critical population densities, without impeding normal bacterial growth. EcN CXCL13 release promoted B cell and splenocyte migration in vitro and high tumor expression after delivery into mouse bladders. In orthotopic bladder cancer models, EcN-colonized tumors selectively elicited GC responses in tdLNs. EcN boosted anti-PD-1-induced antitumor activity, tumor-specific antibody responses, and long-term survival in “cold” advanced bladder cancer models, and generated immune memory. Efficacy depended on CD8⁺ T and CD4⁺ TFH cells.

Contributed by Paula Hochman

Rouanne et al. engineered probiotic E.coli (EcN) using a synchronized lysis integrated circuit to release human CXCL13 at critical population densities, without impeding normal bacterial growth. EcN CXCL13 release promoted B cell and splenocyte migration in vitro and high tumor expression after delivery into mouse bladders. In orthotopic bladder cancer models, EcN-colonized tumors selectively elicited GC responses in tdLNs. EcN boosted anti-PD-1-induced antitumor activity, tumor-specific antibody responses, and long-term survival in “cold” advanced bladder cancer models, and generated immune memory. Efficacy depended on CD8⁺ T and CD4⁺ TFH cells.

Contributed by Paula Hochman

ABSTRACT: The intratumoral microbiome has recently emerged as a potential hallmark of cancer, with implications for response or resistance to therapy. Bacteria can either promote or inhibit cancer growth. However, intratumoral bacteria can also be engineered using synthetic biology to remodel the tumor microenvironment. Here, we engineered the probiotic bacterium Escherichia coli Nissle 1917 (EcN) to express the human chemokine CXCL13 (C-X-C motif chemokine ligand 13), a critical component of germinal center (GC) formation. Antibody affinity maturation and class switching are fundamental aspects of adaptive immune response. Both occur primarily in the GCs of secondary lymphoid organs for defense against pathogens. Immune checkpoint blockade (ICB) efficacy is primarily driven by T cells; however, recent studies in mice and humans have shown that humoral immune responses act as critical partners for ICB-mediated antitumor activity. Using orthotopic models of bladder cancer, intravesically delivered engineered CXCL13-expressing EcN colonized bladder tumors and elicited GC responses in bladder tumor-draining lymph nodes after intravesical delivery. When combined with programmed cell death protein 1 (PD-1) blockade, engineered EcN improved antitumor activity in two aggressive, fast-growing, and immunologically cold orthotopic mouse models of bladder cancer. Mechanistically, this antitumor effect was dependent on the presence of CD8(+) T cells and CD4(+) T follicular helper cells; combination therapy increased tumor-specific antibody responses and promoted long-term survival and protective immunity upon tumor rechallenge. Thus, we demonstrate that synthetically engineered CXCL13-expressing EcN can enhance the efficacy of PD-1 checkpoint blockade immunotherapy by amplifying tumor-specific humoral immunity.

Author Info: (1) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA.

Author Info: (1) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA. (2) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA. (3) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA. (4) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. (5) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA. (6) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. (7) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. (8) Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA. (9) Department of Urology, Columbia University, New York, NY 10032, USA. (10) Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA. Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA. Data Science Institute, Columbia University, New York, NY 10027, USA. (11) Department of Microbiology & Immunology, Columbia University, New York, NY 10032, USA. Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY 10032, USA.

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