Journal Articles

Pancreatic cancer-restricted cryptic antigens are targets for T cell recognition Featured  

Ely, Kulstad, et al. developed an immunopeptidomics pipeline to investigate noncanonical HLA-I-bound peptides (ncHLAp) in patient-derived organoids of pancreatic ductal carcinoma (PDAC). A subset (~30%) of detected ncHLAp were cancer-restricted (CR), which were partially shared between patients. The detected CR ncHLAp were immunogenic. Engineered CR ncHLAp-specific TCR-T cells could detect endogenous levels of expression of presented peptides in vitro, and high-avidity TCR-T cells were found to temporarily delay tumor growth in vivo.

Ely, Kulstad, et al. developed an immunopeptidomics pipeline to investigate noncanonical HLA-I-bound peptides (ncHLAp) in patient-derived organoids of pancreatic ductal carcinoma (PDAC). A subset (~30%) of detected ncHLAp were cancer-restricted (CR), which were partially shared between patients. The detected CR ncHLAp were immunogenic. Engineered CR ncHLAp-specific TCR-T cells could detect endogenous levels of expression of presented peptides in vitro, and high-avidity TCR-T cells were found to temporarily delay tumor growth in vivo.

ABSTRACT: Translation of the noncoding genome in cancer can generate cryptic (noncanonical) peptides capable of presentation by human leukocyte antigen class I (HLA-I); however, the cancer specificity and immunogenicity of noncanonical HLA-I-bound peptides (ncHLAp) are incompletely understood. Using high-resolution immunopeptidomics, we discovered that cryptic peptides are abundant in the pancreatic cancer immunopeptidome. Approximately 30% of ncHLAp exhibited cancer-restricted translation, and a substantial subset were shared among patients. Cancer-restricted ncHLAp displayed robust immunogenic potential in a sensitive ex vivo T cell priming platform. ncHLAp-reactive, T cell receptor-redirected T cells exhibited tumoricidal activity against patient-derived pancreatic cancer organoids. These findings demonstrate that pancreatic cancer harbors cancer-restricted ncHLAp that can be recognized by cytotoxic T cells. Future therapeutic strategies for pancreatic cancer, and potentially other solid tumors, may include targeting cryptic antigens.

Author Info: (1) Koch Institute at MIT, Cambridge, MA, USA. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. (2) Koch Institute at MIT, Cambridge, MA, USA. Dana

Author Info: (1) Koch Institute at MIT, Cambridge, MA, USA. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. (2) Koch Institute at MIT, Cambridge, MA, USA. Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (3) Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. (4) Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (5) The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (6) Dana-Farber Cancer Institute, Boston, MA, USA. (7) The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (8) Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (9) The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (10) Dana-Farber Cancer Institute, Boston, MA, USA. (11) Program in Immunology, Fred Hutchinson Cancer Center, Seattle, WA, USA. (12) Dana-Farber Cancer Institute, Boston, MA, USA. (13) Koch Institute at MIT, Cambridge, MA, USA. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. (14) Dana-Farber Cancer Institute, Boston, MA, USA. (15) Dana-Farber Cancer Institute, Boston, MA, USA. Harvard Medical School, Boston, MA, USA. Hale Family Center for Pancreatic Cancer Research at DFCI, Boston, MA, USA. (16) Department of Health Technology, Technical University of Denmark, Kongens Lyngby, Denmark. (17) The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (18) Koch Institute at MIT, Cambridge, MA, USA. (19) Koch Institute at MIT, Cambridge, MA, USA. (20) Dana-Farber Cancer Institute, Boston, MA, USA. Harvard Medical School, Boston, MA, USA. Hale Family Center for Pancreatic Cancer Research at DFCI, Boston, MA, USA. (21) Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. Hale Family Center for Pancreatic Cancer Research at DFCI, Boston, MA, USA. (22) Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. (23) Program in Immunology, Fred Hutchinson Cancer Center, Seattle, WA, USA. Division of Medical Oncology, Department of Medicine, University of Washington, Seattle, WA, USA. Department of Immunology, University of Washington, Seattle, WA, USA. (24) Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. Hale Family Center for Pancreatic Cancer Research at DFCI, Boston, MA, USA. (25) Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (26) The Broad Institute of MIT and Harvard, Cambridge, MA, USA. (27) Koch Institute at MIT, Cambridge, MA, USA. Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. (28) Koch Institute at MIT, Cambridge, MA, USA. Dana-Farber Cancer Institute, Boston, MA, USA. The Broad Institute of MIT and Harvard, Cambridge, MA, USA. Harvard Medical School, Boston, MA, USA. Hale Family Center for Pancreatic Cancer Research at DFCI, Boston, MA, USA.

Immunotherapy-related cognitive impairment after CAR T cell therapy in mice Spotlight 

Geraghty and Acosta-Alvarez et al. investigated mechanisms underlying impaired cognition following CAR T cell therapy for CNS and non-CNS tumors. In several mouse models, CAR T cells induced neuroinflammation with white matter microglial reactivity, increases in levels of CSF cytokines and chemokines, and dysregulation of oligodendroglial cells and hippocampal neurogenesis. RNAseq analysis of frontal lobes from CAR T cell-treated patients with brain stem tumors confirmed microglial and oligodendrocyte reactivity. In mice, transient depletion of microglia or CCR3 blockade rescued oligodendroglial deficits and improved cognitive performance.

Contributed by Katherine Turner

Geraghty and Acosta-Alvarez et al. investigated mechanisms underlying impaired cognition following CAR T cell therapy for CNS and non-CNS tumors. In several mouse models, CAR T cells induced neuroinflammation with white matter microglial reactivity, increases in levels of CSF cytokines and chemokines, and dysregulation of oligodendroglial cells and hippocampal neurogenesis. RNAseq analysis of frontal lobes from CAR T cell-treated patients with brain stem tumors confirmed microglial and oligodendrocyte reactivity. In mice, transient depletion of microglia or CCR3 blockade rescued oligodendroglial deficits and improved cognitive performance.

Contributed by Katherine Turner

ABSTRACT: Immunotherapies have revolutionized cancer care for many tumor types, but their potential long-term cognitive impacts are incompletely understood. Here, we demonstrated in mouse models that chimeric antigen receptor (CAR) T cell therapy for both central nervous system (CNS) and non-CNS cancers impaired cognitive function and induced a persistent CNS immune response characterized by white matter microglial reactivity, microglial chemokine expression, and elevated cerebrospinal fluid (CSF) cytokines and chemokines. Consequently, oligodendroglial homeostasis and hippocampal neurogenesis were disrupted. Single-nucleus sequencing studies of human frontal lobe from patients with or without previous CAR T cell therapy for brainstem tumors confirmed reactive states of microglia and oligodendrocytes following treatment. In mice, transient microglial depletion or CCR3 chemokine receptor blockade rescued oligodendroglial deficits and cognitive performance in a behavioral test of attention and short-term memory function following CAR T cell therapy. Taken together, these findings illustrate targetable neural-immune mechanisms underlying immunotherapy-related cognitive impairment.

Author Info: (1) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.

Author Info: (1) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA. (2) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (3) Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA. (4) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (5) Neuroscience Institute, NYU Grossman School of Medicine, New York, NY 10016, USA. (6) Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA. (7) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (8) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (9) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (10) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (11) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (12) Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA. (13) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (14) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (15) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (16) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (17) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (18) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (19) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (20) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (21) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA. (22) Brain immunology and Glia (BIG) Center and Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63130, USA. (23) Brain immunology and Glia (BIG) Center and Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63130, USA. (24) Brain immunology and Glia (BIG) Center and Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63130, USA. (25) Neuroscience Institute, NYU Grossman School of Medicine, New York, NY 10016, USA; Department of Neuroscience and Physiology, NYU Grossman School of Medicine, New York, NY 10016, USA; Department of Ophthalmology, NYU Grossman School of Medicine, New York, NY 10016, USA; Parekh Center for Interdisciplinary Neurology, NYU Grossman School of Medicine, New York, NY 10016, USA. (26) Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA; Center for Cancer Cellular Therapy, Stanford School of Medicine, Stanford, CA 94305, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA. (27) Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA; Center for Cancer Cellular Therapy, Stanford School of Medicine, Stanford, CA 94305, USA. (28) Department of Neurology and Neurosciences, Stanford School of Medicine, Stanford, CA 94305, USA; Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA; Center for Cancer Cellular Therapy, Stanford School of Medicine, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA. Electronic address: mmonje@stanford.edu.

Spatial and multiomics analysis of human and mouse lung adenocarcinoma precursors reveals TIM-3 as a putative target for precancer interception Spotlight 

Zhu et al. demonstrated a coordinated interplay between innate and adaptive immunity during the initiation and progression of LUAD pre-cancers, revealing stage-specific remodeling of the tumor microenvironment, including shifts in macrophage polarization and T lymphocyte differentiation. TIM-3 expression increased at early phases of carcinogenesis in human LUAD and in 5 genetically distinct LUAD mouse models. Anti-TIM-3 treatment shifted M1/M2 macrophage polarization, enhanced antigen presentation by DCs, increased T cell activation and cytotoxicity, and decreased tumor burden at pre-cancer, but not advanced cancer stages.

Contributed by Shishir Pant

Zhu et al. demonstrated a coordinated interplay between innate and adaptive immunity during the initiation and progression of LUAD pre-cancers, revealing stage-specific remodeling of the tumor microenvironment, including shifts in macrophage polarization and T lymphocyte differentiation. TIM-3 expression increased at early phases of carcinogenesis in human LUAD and in 5 genetically distinct LUAD mouse models. Anti-TIM-3 treatment shifted M1/M2 macrophage polarization, enhanced antigen presentation by DCs, increased T cell activation and cytotoxicity, and decreased tumor burden at pre-cancer, but not advanced cancer stages.

Contributed by Shishir Pant

ABSTRACT: How tumor microenvironment shapes lung adenocarcinoma (LUAD) precancer evolution remains poorly understood. Spatial immune profiling of 114 human LUAD and LUAD precursors reveals a progressive increase of adaptive response and a relative decrease of innate immune response as LUAD precursors progress. The immune evasion features align the immune response patterns at various stages. TIM-3-high features are enriched in LUAD precancers, which decrease in later stages. Furthermore, single-cell RNA sequencing (scRNA-seq) and spatial immune and transcriptomics profiling of LUAD and LUAD precursor specimens from 5 mouse models validate high TIM-3 features in LUAD precancers. In vivo TIM-3 blockade at precancer stage, but not at advanced cancer stage, decreases tumor burden. Anti-TIM-3 treatment is associated with enhanced antigen presentation, T cell activation, and increased M1/M2 macrophage ratio. These results highlight the coordination of innate and adaptive immune response/evasion during LUAD precancer evolution and suggest TIM-3 as a potential target for LUAD precancer interception.

Author Info: (1) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Genomic Medicine, The University of Te

Author Info: (1) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (2) Institute for Data Science in Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (3) Department of Imaging Physics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (4) Department of Medicine, Baylor College of Medicine, Houston, TX, USA. (5) Clinical Research Center in Hiroshima, Hiroshima University Hospital, Hiroshima, Japan. (6) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (7) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (8) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (9) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (10) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (11) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (12) Department of Pathology, NYU Langone Health, New York, NY, USA. (13) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (14) Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (15) Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (16) Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (17) Department of Imaging Physics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (18) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (19) Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (20) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (21) Department of Translational Molecular Pathology and Sheikn Ahmed Center for Pancreatic Cancer Research, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (22) Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (23) Department of Systems Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (24) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (25) Department of Cancer Systems Imaging, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (26) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (27) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (28) Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (29) Department of Medicine, NYU Grossman School of Medicine, New York, NY, USA. (30) Department of Medicine, NYU Grossman School of Medicine, New York, NY, USA; Cardiothoracic Surgery, NYU Grossman School of Medicine, New York, NY, USA. (31) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (32) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (33) Department of Cellular and Molecular Medicine, UC San Diego, La Jolla, CA, USA. (34) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (35) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (36) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (37) Department of Pathology Informatics, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan. (38) Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (39) Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (40) Department of Cardiothoracic Surgery, NYU Langone Health, New York, NY, USA. (41) Institute of Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, CA, USA. (42) Department of Clinical Cancer Prevention, The University of Texas MD Anderson Cancer Center, Houston, CA, USA. (43) Department of Medicine, Baylor College of Medicine, Houston, TX, USA. (44) Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA, USA. (45) Pathology & Laboratory Medicine, and Bioinformatics, Boston University, Boston, MA, USA. (46) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (47) UC San Diego School of Medicine, La Jolla, CA, USA. (48) Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (49) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (50) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (51) Department of Imaging Physics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. (52) Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. Electronic address: jzhang20@mdanderson.org.

An in situ engineered chimeric IL-2 receptor potentiates the tumoricidal activity of proinflammatory CAR macrophages in renal cell carcinoma Spotlight 

Circular RNAs encoding a CAR and a chimeric IL-2 receptor (IL-2R signaling through an intracellular TLR4 domain) were encapsulated in LNPs for targeted CAR macrophage engineering, leading to significant M1 polarization, tumor cell phagocytosis, and T cell cytotoxicity in vitro. The LNPs were incorporated into an HA/gelatin hydrogel with IL-2, which was injected into the kidney capsule to promote tumor control in orthotopic, resection, and patient-derived xenograft RCC models. While i.t. CAR macrophages were detectable for ~2 weeks, the treatment promoted long-term survival, in part via enhanced M1 polarization and T cell infiltration.

Contributed by Morgan Janes

Circular RNAs encoding a CAR and a chimeric IL-2 receptor (IL-2R signaling through an intracellular TLR4 domain) were encapsulated in LNPs for targeted CAR macrophage engineering, leading to significant M1 polarization, tumor cell phagocytosis, and T cell cytotoxicity in vitro. The LNPs were incorporated into an HA/gelatin hydrogel with IL-2, which was injected into the kidney capsule to promote tumor control in orthotopic, resection, and patient-derived xenograft RCC models. While i.t. CAR macrophages were detectable for ~2 weeks, the treatment promoted long-term survival, in part via enhanced M1 polarization and T cell infiltration.

Contributed by Morgan Janes

ABSTRACT: Chimeric antigen receptor macrophage (CAR-M) therapy has shown great promise in solid malignancies; however, the phenotypic re-domestication of CAR-Ms in the immunosuppressive tumor niche restricts their antitumor immunity. We here report an in situ engineered chimeric interleukin (IL)-2 signaling receptor (CSR) for controllably manipulating the proinflammatory phenotype of CAR-Ms, augmenting their sustained tumoricidal immunity. Specifically, our in-house-customized lipid nanoparticles efficiently introduce dual circular RNAs into macrophages to generate CSR-functionalized CAR-Ms. The intracellular inflammatory signaling pathway of CAR-Ms can be stimulated with the IL-2 therapeutic via the synthetic IL-2 receptor, which induces the antitumor phenotype shifting of CAR-Ms. Moreover, hydrogel-mediated combinatory treatment with lipid nanoparticles and IL-2 remodels the immunosuppressive tumor microenvironment and promotes tumor regression in renal carcinoma animal models. In summary, our findings establish that the proinflammatory phenotype of CAR-Ms can be modulated by a synthetic IL-2 receptor, benefiting the antitumor immunotherapy of CAR-Ms with broad application in other solid malignancies.

Author Info: (1) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technol

Author Info: (1) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. wjing1@sdu.edu.cn. (2) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (3) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (4) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (5) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (6) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (7) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (8) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (9) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (10) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (11) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (12) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (13) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (14) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (15) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (16) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (17) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (18) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. (19) Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Kowloon, China. (20) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. shibenkangsdu@163.com. (21) Department of Urology, Qilu Hospital, Cheeloo College of Medicine; Shandong Key Laboratory of Targeted Drug Delivery and Advanced Pharmaceutics, NMPA Key Laboratory for Technology Research and Evaluation of Drug Products and Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmaceutics, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China. xinyijiang@sdu.edu.cn.

Impaired T cell and neoantigen retention in time-serial analysis of metastatic non-small cell lung cancer in patients unresponsive to TIL cell therapy Spotlight 

Wang et al. performed multidimensional analysis on longitudinal tumor and blood samples in a clinical trial of TIL therapy in 16 patients with metastatic NSCLC. In patients that did not clinically benefit, tumor neoantigen-reactive clonotypes were less cytolytic, expressed a dysfunctional program, and lacked stem/memory-like self-renewal. Further, loss of infused cells or of neoantigen-reactive peripheral T cell clonotypes over time was associated with the onset of progressive disease. Subclonal neoantigens that were previously targeted by infused TILs were absent from tumor cell genomes upon progression, suggesting adaptive resistance.

Contributed by Lauren Hitchings

Wang et al. performed multidimensional analysis on longitudinal tumor and blood samples in a clinical trial of TIL therapy in 16 patients with metastatic NSCLC. In patients that did not clinically benefit, tumor neoantigen-reactive clonotypes were less cytolytic, expressed a dysfunctional program, and lacked stem/memory-like self-renewal. Further, loss of infused cells or of neoantigen-reactive peripheral T cell clonotypes over time was associated with the onset of progressive disease. Subclonal neoantigens that were previously targeted by infused TILs were absent from tumor cell genomes upon progression, suggesting adaptive resistance.

Contributed by Lauren Hitchings

ABSTRACT: Cell therapy with tumor-infiltrating lymphocytes (TILs) has yielded durable responses for multiple cancer types, but the causes of therapeutic resistance remain largely unknown. Here multidimensional analysis was performed on time-serial tumor and blood in a lung cancer TIL therapy trial. Using T cell receptor sequencing on both functionally expanded T cells and neoantigen-loaded tetramer-sorted T cells, we identified tumor antigen-specific T cell receptors. We then mapped clones into individual transcriptomes and found that tumor-reactive clonotypes expressed a dysfunctional program and lacked stem-like features among patients who lacked clinical benefit. Tracking tumor-reactive clonotypes over time, decay of antigen-reactive peripheral T cell clonotypes was associated with the emergence of progressive disease. Further, subclonal neoantigens previously targeted by infused T cells were subsequently absent within tumors at progression, suggesting potential adaptive resistance. Our findings suggest that targeting clonal antigens and circumventing dysfunctional states may be important for conferring clinical responses to TIL therapy.

Author Info: (1) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. chao.wang@moffitt.org. (2) Department of Biostatistics and Bioinformatics, H. Lee M

Author Info: (1) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. chao.wang@moffitt.org. (2) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (3) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (4) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (5) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (6) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (7) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (8) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (9) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (10) Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK. (11) Center for Immunotherapy and Precision Immuno-oncology, Cleveland Clinic Lerner Research Institute, Cleveland, OH, USA. (12) Center for Immunotherapy and Precision Immuno-oncology, Cleveland Clinic Lerner Research Institute, Cleveland, OH, USA. (13) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. Department of Integrative Immunobiology, Duke School of Medicine, Durham, NC, USA. Duke Cancer Institute, Duke University School of Medicine, Durham, NC, USA. (14) Department of Anatomic Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (15) Department of Anatomic Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (16) Proteomics & Metabolomics Core Facility, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (17) Molecular Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (18) Immune and Cellular Therapy Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (19) Immune and Cellular Therapy Program, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (20) Molecular Genomics Core Facility, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (21) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. Department of Radiation Oncology, Immunology, Cancer Biology, Mayo Clinic Alix College of Medicine & Health Sciences, Jacksonville, FL, USA. (22) Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (23) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (24) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. Department of Integrative Immunobiology, Duke School of Medicine, Durham, NC, USA. Duke Cancer Institute, Duke University School of Medicine, Durham, NC, USA. (25) Duke Cancer Institute, Duke University School of Medicine, Durham, NC, USA. (26) Department of Thoracic Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. (27) Department of Immunology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. ben.creelan@moffitt.org. Department of Thoracic Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. ben.creelan@moffitt.org.

Somatic mutations in HLA class genes and antigen presenting molecules in malignant glioma Spotlight 

Schulte, Peter, and Rosenberger et al. evaluated IDH-mutant and wild-type gliomas for mutations in 16 genes encoding proteins involved in antigen presentation (HLAs and HLA-related proteins) using a bioinformatics pipeline specifically designed to detect mutations in these highly polymorphic genetic regions. This effectively identified somatic mutations in genes encoding HLA-II, nonclassical HLA genes, TAP1/2, and B2M. 3D modeling of non-synonomous mutations in TAP1 and B2M demonstrated that such mutations could disrupt antigen presentation, potentially mediating escape from T cells. Mutations were found to be more frequent in recurrent glioblastomas.

Contributed by Lauren Hitchings

Schulte, Peter, and Rosenberger et al. evaluated IDH-mutant and wild-type gliomas for mutations in 16 genes encoding proteins involved in antigen presentation (HLAs and HLA-related proteins) using a bioinformatics pipeline specifically designed to detect mutations in these highly polymorphic genetic regions. This effectively identified somatic mutations in genes encoding HLA-II, nonclassical HLA genes, TAP1/2, and B2M. 3D modeling of non-synonomous mutations in TAP1 and B2M demonstrated that such mutations could disrupt antigen presentation, potentially mediating escape from T cells. Mutations were found to be more frequent in recurrent glioblastomas.

Contributed by Lauren Hitchings

ABSTRACT: Immune evasion is a hallmark of gliomas, yet the genetic mechanisms by which tumors escape immune surveillance remain incompletely understood. In this study, we systematically examined the presence of somatic mutations in human leukocyte antigen (HLA) genes and genes encoding proteins involved in antigen-presentation across isocitrate dehydrogenase wild-type (IDHwt) and mutant (IDHmut) gliomas using targeted next-generation sequencing (NGS). To address the challenges associated with detecting somatic mutations in these highly polymorphic and complex regions of the genome, we applied a combination of short-read and long-read sequencing techniques, extended the genetic region of interest (exons and introns), and applied a tailored bioinformatics analysis pipeline, which enabled an accurate evaluation of comprehensive sequencing data. Our analysis identified mutations in HLA class II and non-classical HLA genes as well as genes associated with antigen presentation, such as TAP1/2 and B2M. Three-dimensional modeling of individual mutations simulated the potential impact of somatic mutations in TAP1 and B2M on the encoded protein configuration. The presence of somatic mutations supports the role of antigen-presenting genes in the pathophysiology and potential immune escape of gliomas. Our data demonstrated an increased frequency of such mutations in recurrent glioblastoma (GBM), potentially resulting from a positive selection or mutagenic enrichment of tumor cells during tumor progression. Taken together, this research generates new insights and hypotheses for the functional analysis and optimization of immunotherapy strategies for gliomas, which may guide personalized treatment paradigms.

Author Info: (1) Heinrich Heine University DŸsseldorf, DŸsseldorf, Germany. (2) Stefan-Morsch-Stiftung, Birkenfeld, Rheinland-Pfalz, Germany. (3) Institute of Human Genetics, University Medical

Author Info: (1) Heinrich Heine University DŸsseldorf, DŸsseldorf, Germany. (2) Stefan-Morsch-Stiftung, Birkenfeld, Rheinland-Pfalz, Germany. (3) Institute of Human Genetics, University Medical Center Hamburg-Eppendorf, Hamburg, Germany, Hamburg, Hamburg, Germany. (4) Stefan Morsch Stiftung, Birkenfeld, Germany, Germany. (5) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (6) Laboratory for Brain Tumor Biology, Department of Neurosurgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany, Germany. (7) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (8) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (9) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (10) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (11) Hubertus Wald University Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (12) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (13) Heinrich Heine University DŸsseldorf, DŸsseldorf, Germany. (14) University Medical Center Hamburg-Eppendorf, Hamburg, Hamburg, Germany. (15) University Medical Center Hamburg-Eppendorf, Germany. (16) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (17) University Medical Center Hamburg-Eppendorf, Hamburg, Germany. (18) Institute of Medical Microbiology and Hospital Hygiene, University Hospital DŸsseldorf, Heinrich Heine University DŸsseldorf, DŸsseldorf, Germany, Germany. (19) University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

Improved overall survival in an anti-PD-L1 treated cohort of newly diagnosed glioblastoma patients is associated with distinct immune, mutation, and gut microbiome features: a single arm prospective phase I/II trial Spotlight 

Weathers et al. reported that the concurrent use of atezolizumab in combination with radiation and TMZ was safe, and demonstrated efficacy consistent with published trials for newly diagnosed GBM. Patients harboring an EGFR mutation had a relatively worse mOS compared to those with a PTEN mutation. Gene set enrichment analysis identified multiple immune-related gene sets enriched in patients with longer OS, suggesting that a subset of GBM tumors exhibited higher immune infiltration. These immune-infiltrated tumors showed enrichment for the mesenchymal GBM subtype, distinct fecal microbiome profiles, and specific immune cell populations.

Contributed by Shishir Pant

Weathers et al. reported that the concurrent use of atezolizumab in combination with radiation and TMZ was safe, and demonstrated efficacy consistent with published trials for newly diagnosed GBM. Patients harboring an EGFR mutation had a relatively worse mOS compared to those with a PTEN mutation. Gene set enrichment analysis identified multiple immune-related gene sets enriched in patients with longer OS, suggesting that a subset of GBM tumors exhibited higher immune infiltration. These immune-infiltrated tumors showed enrichment for the mesenchymal GBM subtype, distinct fecal microbiome profiles, and specific immune cell populations.

Contributed by Shishir Pant

ABSTRACT: This phase I/II trial aims to evaluate the efficacy of concurrent atezolizumab with radiation therapy and temozolomide (TMZ) followed by adjuvant atezolizumab and TMZ in newly diagnosed glioblastoma (GBM) patients and to identify pre-treatment correlates with outcome (N = 60). Trial number: NCT03174197. The primary outcome was overall survival (OS) whereas secondary outcomes were retrospective global-omics analyses to identify pre-treatment immune and genetic tumor features that correlated with survival. Concurrent use of atezolizumab with radiation and TMZ demonstrated OS in line with published trials for newly diagnosed GBM. Tumor genomic (WES and/or targeted NGS panel), transcriptomic (RNAseq) and tissue microenvironment imaging, as well as fecal metagenomic sequencing were conducted. Gene set enrichment analysis of tumors identified multiple immune-based transcriptomic programs to distinguish patients with longer versus shorter survival (p ≤ 0.01). GBM immune enrichment was highly associated with the pre-treatment tumor mesenchymal subtype and patient gastrointestinal bacterial taxa profile.

Author Info: (1) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. sweathers@mdanderson.org. (2) Genomic Med

Author Info: (1) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. sweathers@mdanderson.org. (2) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (3) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (4) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (5) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (6) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (7) Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (8) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (9) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (10) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (11) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (12) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (13) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (14) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (15) Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (16) Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (17) Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (18) Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (19) Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (20) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (21) Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (22) Genentech, Inc 1 DNA Way, San Francisco, CA, 94080, USA. (23) Genentech, Inc 1 DNA Way, San Francisco, CA, 94080, USA. (24) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (25) Genomic Medicine, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. (26) Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, 1515 Holcombe Blvd, Houston, TX, 77030, USA. john.degroot@ucsf.edu.

cGAS mRNA-Based Immune Agonist Promotes Vaccine Responses and Antitumor Immunity Spotlight 

Qu, Li et al. generated lipid nanoparticles (LNPs) encapsulating mRNA for cGAS, the innate immune sensor of cytoplasmic dsDNA. Human cGAS was mutated (K187N/L195R) to enhance its activation of the STING-dependent IFN response; induceAPC maturation, antigen uptake, and presentation; and upregulate expression of MHC-I, MHC-II, and costimulatory signals in vitro and in mice. Treating mice with mutated hcGASmRNA-LNPs induced strong innate and adaptive immune responses that boosted the potency of mRNA, protein, and tumor vaccines, and also synergized with IFNγ treatment to act directly on tumor cells to reduce tumor volume and increase host survival.

Contributed by Paula Hochman

Qu, Li et al. generated lipid nanoparticles (LNPs) encapsulating mRNA for cGAS, the innate immune sensor of cytoplasmic dsDNA. Human cGAS was mutated (K187N/L195R) to enhance its activation of the STING-dependent IFN response; induceAPC maturation, antigen uptake, and presentation; and upregulate expression of MHC-I, MHC-II, and costimulatory signals in vitro and in mice. Treating mice with mutated hcGASmRNA-LNPs induced strong innate and adaptive immune responses that boosted the potency of mRNA, protein, and tumor vaccines, and also synergized with IFNγ treatment to act directly on tumor cells to reduce tumor volume and increase host survival.

Contributed by Paula Hochman

ABSTRACT: mRNA vaccines are a potent tool for immunization against viral diseases and cancer. However, the lack of a vaccine adjuvant limits the efficacy of these treatments. In this study, we used cGAS mRNA, which encodes the DNA innate immune sensor, complexed with lipid nanoparticles (LNP), to boost the immune response. By introducing specific mutations in human cGAS mRNA (hcGASK187N/L195R), we significantly enhanced cGAS activity, resulting in a more potent and sustained stimulator of interferon gene (STING)-mediated IFN response. cGAS mRNA-LNPs exhibited stimulatory effects on maturation, antigen engulfment, and antigen presentation by antigen-presenting cells, both in vitro and in vivo. Moreover, the hcGASK187N/L195R mRNA-LNP combination demonstrated a robust adjuvant effect and amplified the potency of mRNA and protein vaccines, which was a result of strong humoral and cell-mediated responses. Remarkably, the hcGASK187N/L195R mRNA-LNP complex, either alone or in combination with antigens, demonstrated exceptional efficacy in eliciting antitumor immunity. In addition to its immune-boosting properties, hcGASK187N/L195R mRNA-LNP exerted antitumor effects with IFN_ directly on tumor cells, further promoting tumor restriction. In conclusion, we developed a cGAS mRNA-based immunostimulatory adjuvant compatible with various vaccine forms to boost the adaptive immune response and cancer immunotherapies.

Author Info: (1) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Cen

Author Info: (1) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (2) Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. Department of Radiology, The First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China. (3) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (4) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (5) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (6) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (7) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (8) Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (9) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (10) The First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China. (11) The First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China. (12) Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (13) The First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China. (14) Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. Department of Radiology, The First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China. (15) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. (16) Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China. Department of Radiology, The First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China. (17) National Key Laboratory of Immune Response and Immunotherapy, Department of General Surgery, The First Affiliated Hospital of University of Science and Technology of China, Center for Advanced Interdisciplinary Science & Biomedicine IHM, Division of Life Sciences & Medicine, University of Science and Technology of China, Hefei, China. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China.

CAR Binders Affect CAR T-cell Tonic Signaling, Durability, and Sensitivity to Target Spotlight 

The murine anti-CD19 scFv FMC63 is employed in FDA-approved CAR T cell therapies. To reduce the potential for immunogenicity, Shukla et al. screened a library of fully human anti-CD19 scFvs for expression, functionality (CD19 binding and cytokine production), and tonic signaling. The best binders killed target cells in vitro, controlled tumors in vivo, and responded to low antigen densities. The top fully human scFv bound a different region of CD19 than FMC63, had weaker CD19 affinity (25nM vs. 0.6nM), and demonstrated comparable or superior in vitro tumor killing, CD19-low responses, and in vivo efficacy to FMC63, and is currently being tested in the clinic.

Contributed by Alex Najibi

The murine anti-CD19 scFv FMC63 is employed in FDA-approved CAR T cell therapies. To reduce the potential for immunogenicity, Shukla et al. screened a library of fully human anti-CD19 scFvs for expression, functionality (CD19 binding and cytokine production), and tonic signaling. The best binders killed target cells in vitro, controlled tumors in vivo, and responded to low antigen densities. The top fully human scFv bound a different region of CD19 than FMC63, had weaker CD19 affinity (25nM vs. 0.6nM), and demonstrated comparable or superior in vitro tumor killing, CD19-low responses, and in vivo efficacy to FMC63, and is currently being tested in the clinic.

Contributed by Alex Najibi

ABSTRACT: Patients can develop human anti-mouse immune responses against CD19-specific chimeric antigen receptor (CAR) T cells due to the use of a murine CD19-specific single-chain variable fragment to redirect T cells. We screened a yeast display library to identify an array of fully human CD19 single-chain variable fragment binders and performed a series of studies to select the most promising fully human CAR. We observed significant differences in the ability of CARs employing these CD19 binders to be expressed on the cell surface, induce tonic signaling, redirect T-cell function, mediate tumor killing, recognize lower levels of CD19 antigen, and maintain function upon continuous antigen exposure. From this initial analysis, CAR T cells using two binders (42 and 52) were selected for additional studies. Although CAR T cells using both binders controlled tumor growth well in vivo, we advanced a CAR construct using binder 42 for more advanced preclinical testing because of its greater similarity to binders based on the antibody FMC63, which is the murine antibody underlying four FDA-approved CD19-specific CAR T-cell therapies, and ability to robustly respond to tumors expressing lower levels of CD19. We found that this binder uniquely bound CD19 using distinct contact residues than FMC63 and with _40-fold lower affinity. CARs using binder 42 were non-inferior to those using the FMC63 binder in a mouse model of acute lymphoblastic leukemia, indicating that CAR T cells using binder 42 should be considered for clinical use.

Author Info: (1) Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania. Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania.

Author Info: (1) Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania. Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. (2) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. (3) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. (4) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. (5) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. (6) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. (7) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. (8) Tmunity Therapeutics/Kite Pharma, Philadelphia, Pennsylvania. (9) Tmunity Therapeutics/Kite Pharma, Philadelphia, Pennsylvania. (10) Tmunity Therapeutics/Kite Pharma, Philadelphia, Pennsylvania. (11) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. (12) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. Parker Institute for Cancer Immunotherapy, University of Pennsylvania, Philadelphia, Pennsylvania. (13) Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania. (14) Department of Microbiology, University of Pennsylvania, Philadelphia, Pennsylvania. Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania.

A dendritic cell-like transition of T cells is associated with spontaneous remission of adult T-cell leukemia-lymphoma Spotlight 

Watanabe et al. analyzed ATL cells and cells infected with the ATL-causing HTLV-1, and identified a small number (<5% on average) of cells expressing DC-associated molecules (CD14, CD1c, CD11b, CD11c, and CD141), despite being derived from T cells. In a patient with ATL who entered remission after contracting COVID-19, these DC-like T cells increased, along with CTL responses to the HTLV-1 antigen Tax, suggesting these cells may contribute to antigen presentation and spontaneous regression. In an ATL cell line endogenously expressing BATF3, enforced expression of IRF8 and PU.1 increased CD86 expression and peptide presentation of Tax.

Contributed by Lauren Hitchings

Watanabe et al. analyzed ATL cells and cells infected with the ATL-causing HTLV-1, and identified a small number (<5% on average) of cells expressing DC-associated molecules (CD14, CD1c, CD11b, CD11c, and CD141), despite being derived from T cells. In a patient with ATL who entered remission after contracting COVID-19, these DC-like T cells increased, along with CTL responses to the HTLV-1 antigen Tax, suggesting these cells may contribute to antigen presentation and spontaneous regression. In an ATL cell line endogenously expressing BATF3, enforced expression of IRF8 and PU.1 increased CD86 expression and peptide presentation of Tax.

Contributed by Lauren Hitchings

ABSTRACT: Spontaneous remission in patients with various cancers has been reported. Some patients with adult T-cell leukemia-lymphoma (ATL), have experienced spontaneous remission, although mechanisms for this remain unknown. In this study, we analyzed ATL cells and human T-cell leukemia virus type 1 (HTLV-1) infected cells using Cytometry by Time-Of-Flight mass spectrometry (CyTOF). We observed a small number (less than 5% on average) of ATL cells and HTLV-1 infected cells expressed CD14 and other dendritic cell (DC) associated molecules such as CD1c, CD11b, CD11c, and CD141. Single cell analysis revealed that these T cells expressing DC markers also contained rearranged TCR genes, indicating that these cells are indeed derived from T cells. In an ATL patient who entered into remission after contracting coronavirus disease 2019 (COVID-19), the number of DC-like T cells increased, and ELISPOT assay detected CTLs against Tax in accordance with regression of ATL. These findings suggest that DC-like ATL cells acquire antigen-presenting capability, and induce spontaneous remission through enhanced immunity to the virus. Specifically, in an ATL cell line, enforced expression of IRF8 and PU.1 in addition to endogenous BATF3 expression increased CD86 expression and enabled the cells to present Tax peptide antigens to T cells. Collectively, these data indicate that ATL cells acquire antigen-presenting activity when IRF8, PU.1 and BATF3 are expressed, suggesting that transition of a subset of T cells to DC-like T cells can induce immune responses to viral antigens, resulting in spontaneous remission. Thus, the transition of T cells to DC-like T cells is a unique mechanism for spontaneous remission in ATL.

Author Info: (1) Kumamoto University School of Medicine, Kumamoto, Japan. (2) Kumamoto University School of Medicine, Kumamoto, Japan. (3) Kumamoto University School of Medicine, Kumamoto, Japa

Author Info: (1) Kumamoto University School of Medicine, Kumamoto, Japan. (2) Kumamoto University School of Medicine, Kumamoto, Japan. (3) Kumamoto University School of Medicine, Kumamoto, Japan. (4) Kyoto University, Kyoto, Japan. (5) Kumamoto University School of Medicine, Kumamoto, Japan. (6) Kumamoto University, Kumamoto, Japan. (7) Kumamoto University, Kumamoto, Japan. (8) Graduate School of Medical Sciences, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan. (9) Kumamoto University, Kumamoto, Japan.

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