Document text
Principal Investigator: Kai W Wucherpfennig
Organization: BRIGHAM AND WOMEN'S HOSPITAL
Fiscal Year: 2023
Award: $351,997
Funding agency: National Cancer Institute
Abstract
T cells are central effector cells of protective anti-tumor immunity, but little is currently known about these
important immune cells in human GBM. We have generated single-cell RNA-seq data on tumor-infiltrating T cell
populations from GBM patients at initial diagnosis or relapse. We used these full-length RNA-seq data to identify
clonally expanded T cell populations based on their TCRα and β chain sequences and then examined which
genes were overexpressed by such expanded T cells. This analysis highlighted the KLRB1 gene which encodes
the CD161 receptor that was previously shown to inhibit NK cell-mediated cytotoxicity. The CD161 ligand,
CLEC2D, is expressed at the cell surface of human GBM cells. We therefore hypothesize that the CD161 –
CLEC2D pathway inhibits the anti-tumor function of both CD8 and CD4 effector T cell populations in
GBM. Preliminary data show that inactivation of the KLRB1 gene in primary human T cells greatly enhances
their effector function in a humanized mouse model of GBM. Our preliminary data also demonstrate that several
other inhibitory receptors are expressed by substantial populations of GBM-infiltrating T cells, including CD96
and two prostaglandin E2 receptors (EP2 and EP4). Aim 1 will focus on the analysis of tumor-infiltrating T cells
in GBM patients enrolled in the phase 1b NeoVax plus PD-1 antibody trial described in Project 1. These studies
will primarily focus on the expression of inhibitory receptors by T cells and their ligands by tumor cells and myeloid
cells. Expression of inhibitory receptors and their ligands will be examined by 16-color spectral flow cytometry
and single-cell RNA-seq (in collaboration with Cores 1 and 2), with an emphasis on CD161, PD-1, CD96 and
prostaglandin E2 receptors. We will investigate paired tumor samples from the same patient obtained at initial
surgery and relapse in order to determine how expression of these inhibitory receptors and their ligands changes
following immunotherapy with NeoVax plus PD-1 antibody. In collaboration with Project 1, we will also examine
the spatial distribution of T cells that express CD161 and other inhibitory receptors. Aim 2 will investigate the
therapeutic significance of the CD161 – CLEC2D pathway. We will first use a genetic approach to study this
inhibitory receptor by inactivating the KLRB1 gene in primary T cells. Blocking mAbs specific for human CD161
will also be used to examine the therapeutic potential of these findings. We will also examine combination
therapies (collaboration with Projects 2, 4 and Core 3) involving the inhibitory receptors identified by single-cell
RNA-seq in human GBM infiltrating T cells, with a particular focus on CD161, PD-1, CD96 and the prostaglandin
E2 receptors. These studies will significantly advance our understanding of T cell function in GBM and
characterize important inhibitory receptor – ligand interactions that constrain effector T cell function.
Terms: <Antibodies><B7-H1><B7H1><Blocking Antibodies><C Type Lectin Receptors><CD161><CD274><CD8><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><CD8B><CD8B1><CD8B1 gene><Cancers><Cell Body><Cell Communication and Signaling><Cell Function><Cell Line><Cell Mediated Immunology><Cell Process><Cell Signaling><Cell physiology><Cell surface><Cell-Mediated Cytolysis><Cell-Mediated Immunity><Cell-Mediated Lympholysis><CellLine><Cells><Cellular Cytotoxicity><Cellular Function><Cellular Immunity><Cellular Physiology><Cellular Process><Clinical Treatment Moab><Clinical Trials><Clonal Expansion><Clone Cells><Collaborations><Color><Combined Modality Therapy><Cytotoxic cell><Data><Diagnosis><Dinoprostone><EP4><Effector Cell><Epididymal Secretory Protein E4><Expression Signature><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Gene Expression><Gene Expression Profile><Genes><Glial Cell Tumors><Glial Neoplasm><Glial Tumor><Glioblastoma><Glioma><Goals><Grade IV Astrocytic Neoplasm><Grade IV Astrocytic Tumor><Grade IV Astrocytoma><HE4><Human><IFN-Gamma><IFN-g><IFN-γ><IFNG><IFNγ><Immune><Immune Interferon><Immune mediated therapy><Immunes><Immunocompetent><Immunodeficient Mouse><Immunohistochemistry><Immunohistochemistry Cell/Tissue><Immunohistochemistry Staining Method><Immunologically Directed Therapy><Immunosuppression><Immunosuppression Effect><Immunosuppressive Effect><Immunotherapy><Interferon Gamma><Interferon Type II><Intracellular Communication and Signaling><K lymphocyte><KLRB1><KLRB1 gene><Killer Cell Lectin-Like Receptor Subfamily B, Member 1 Gene><LYT3><Label><Length><Ligands><Lymphocyte Cytotoxicity><Lymphocytotoxicity><Lytotoxicity><MHC Receptor><Major Epididymis-Specific Protein E4><Major Histocompatibility Complex Receptor><Malignant Neoplasms><Malignant Tumor><Mediating><Messenger RNA><Mice><Mice Mammals><Modern Man><Molecular><Monoclonal Antibodies><Multimodal Therapy><Multimodal Treatment><Murine><Mus><Myeloid Cells><NK Cells><NKR-P1><NKR-P1A><NKRP1A><Natural Killer Cells><Neuroglial Neoplasm><Neuroglial Tumor><Operative Procedures><Operative Surgical Procedures><PD 1><PD-1><PD-1 antibody><PD-L1><PD1><PD1 antibody><PDL-1><PDL1><PGE2><PGE2 alpha><PGE2alpha><Pathway interactions><Patients><Peptide Vaccines><Peptide-MHC><Peptide-Major Histocompatibility Protein Complex><Peptide/MHC Complex><Phase><Population><Programmed Cell Death 1 Ligand 1><Programmed Death Ligand 1><Prostaglandin E2><Prostaglandin E2 alpha><Prostaglandin E2alpha><Proteins><Putative Protease Inhibitor WAP5><RNA Seq><RNA sequencing><RNAseq><Receptor Protein><Relapse><Role><Sampling><Signal Transduction><Signal Transduction Systems><Signaling><Spatial Distribution><Strains Cell Lines><Subcellular Process><Surgical><Surgical Interventions><Surgical Procedure><T cell infiltration><T-Cell Activation><T-Cell Antigen Receptors><T-Cell Receptor><T-Cells><T-Lymphocyte><T8 Cells><T8 Lymphocytes><Teff cell><Therapeutic><Tumor Cell><Tumor Immunity><WAP Four-Disulfide Core Domain Protein 2><WAP5><WFDC2><WFDC2 gene><Work><activate T cells><anti-PD-1 Ab><anti-PD-1 antibodies><anti-PD-1 monoclonal antibodies><anti-PD1 Ab><anti-PD1 antibodies><anti-PD1 monoclonal antibodies><anti-programmed cell death protein 1 antibodies><anti-programmed death-1 antibody><anti-tumor immunity><antitumor immunity><biological signal transduction><brain tissue><cancer immunity><cancer infiltrating T cells><cell mediated cytotoxicity><combination therapy><combined modality treatment><combined treatment><cultured cell line><cytokine><cytotoxic><cytotoxicity><dJ461P17.6><effector T cell><exhaust><experiment><experimental research><experimental study><experiments><flow cytophotometry><gene expression pattern><gene expression signature><genetic approach><genetic strategy><glial-derived tumor><glioblastoma multiforme><hNKR-P1A><humanized mice><humanized mouse><immune competent><immune suppression><immune suppressive activity><immune suppressive function><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><immunosuppressive activity><immunosuppressive function><immunosuppressive response><implantation><interest><lFN-Gamma><mAbs><mRNA><malignancy><monoclonal Abs><mouse model><multi-modal therapy><multi-modal treatment><murine model><neo-antigen vaccine><neoantigen vaccine><neoplasm/cancer><neoplastic cell><neuroglia neoplasm><neuroglia tumor><overexpress><overexpression><pMHC><participant enrollment><pathway><patient enrollment><population based><programmed cell death 1><programmed cell death ligand 1><programmed cell death protein 1><programmed cell death protein ligand 1><programmed death 1><programs><protein death-ligand 1><receptor><receptor expression><response><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><sle2><social role><spongioblastoma multiforme><surgery><systemic lupus erythematosus susceptibility 2><thymus derived lymphocyte><transcriptional profile><transcriptional signature><transcriptome sequencing><transcriptomic sequencing><tumor><tumor infiltrating T cells>