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Determination of chromatin protein dynamics in CD8 T cells

CD8 T 细胞染色质蛋白动态测定

基本信息

批准号：
10735641
负责人：
Yongqiang Feng
金额：
$ 27.3万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-23 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10735641
关键词：
Acetylation Antibodies Antigens Binding Bioinformatics Biological Biotinylation CD8-Positive T-Lymphocytes CRISPR screen Cell physiology Cells Cellular biology Cellular immunotherapy Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Coupled Cues Enhancers Epigenetic Process FOXP3 gene Gene Expression Gene Expression Regulation Genes Genetic Transcription Genomic Segment Goals Histone Acetylation Histone H3 Immune System Diseases Immune response Immunologics In Situ In Vitro Infiltration Interleukin-2 Investigation Knock-out Libraries Lysine Malignant Neoplasms Mass Spectrum Analysis Mediating Methods Mus Nuclear Nuclear Proteins Outcome Pathway interactions Physiological Play Process Protein Dynamics Proteins Proteomics Receptor Signaling Regulatory T-Lymphocyte Research Role Signal Pathway Signal Transduction System T cell differentiation T cell response T-Cell Development T-Cell Receptor T-Lymphocyte Testing Therapeutic Time Transcriptional Regulation Transducers Tumor Immunity Viral Work anti-tumor immune response cell type chromatin protein comparative cytokine epigenetic profiling epigenetic regulation experimental study extracellular genetic element genome editing immune function improved in vivo inducible gene expression innovation interest novel programs promoter receptor recruit response screening tumor

项目摘要

Determination of chromatin protein dynamics in CD8 T cells CD8 T cells are crucial for antiviral and antitumor immunity. How these cells respond to stimulations and how to manipulate their effector function for therapeutic purposes have been under intensively investigations. It is widely accepted that T cell antigen receptor (TCR) and co-receptor signaling drives almost all the key processes of CD8 T cell differentiation and function via transcriptional and epigenetic regulation of gene expression. Factors modulating TCR signal transduction or target gene expression presumably play important roles in dictating CD8 T cell differentiation and immunological functions. Although proteins directly mediating TCR and co-receptor signal transduction have been well defined, the nuclear proteins modulating target gene expression remain to be fully investigated. Unbiased approaches are required to comprehensively reveal the nuclear proteins regulating TCR and co-receptor dependent gene expression, potentially leading to the identification of novel factors governing CD8 T cell differentiation and function that determine antiviral and antitumor immune response. Recently CRISPR screening offers an unprecedented efficiency to systematically delineate the regulators of CD8 T cell differentiation and function. However, this method does not uncover the mechanistic linkage between candidate factors and signaling pathways and is also severely constrained by available experimental readouts. We reasoned that defining the dynamic proteins of the active gene promoters and enhancers in CD8 T cells upon TCR and co-receptor stimulation would be a solution, because proteins recruited to or depleted at these regions during this process would reveal novel key regulators. To determine the dynamic chromatin proteins at genomic regions of interest, we recently developed a comparative proteomics method using antibody-guided proximity biotinylation in a separate study. In preliminary experiments, we applied this method to mouse primary CD8 T cells and profiled the dynamic proteins at active enhancers and promoters marked by histone H3 lysine 27 acetylation (H3K27ac) upon TCR and co-stimulation, because this signaling appears to act on these genetic elements to control important functional genes in CD8 T cells (such as cytokines and Myc) by coordinating with the histone acetylation pathway. This experiment revealed known TCR signal transducers and a number of novel factors overrepresented or underrepresented. We then performed an in vivo pooled CRISPR screening by knocking out these dynamic proteins and discovered several positive and negative regulators of tumor infiltrating CD8 T cells. On the basis of these results, we propose to further define the chromatin protein dynamics in CD8 T cells and determine their roles in modulating inducible gene expression and CD8 T cell antitumor activity. Overall, we will employ a new proteomics method, developed by us, to determine signal dependent chromatin protein dynamics to unbiasedly identify novel regulators of CD8 T cell function. These factors may serve as new targets to improve CD8 T cell based immunotherapies.

CD8 T细胞染色质蛋白动力学的测定