Determining the role of nuclear envelope reformation proteins in regulating the cGAS/STING innate immune response in cancer

确定核膜重组蛋白在调节癌症 cGAS/STING 先天免疫反应中的作用

基本信息

批准号：
10750669
负责人：
Anthony Wayne Isenhour
金额：
$ 4.77万
依托单位：
YALE UNIVERSITY
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：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10750669
关键词：
Auxins Biochemical CRISPR/Cas technology Cancer Biology Cancer cell line Cell Line Cellular biology Chromatin Chromosomes Clupeidae Complex Cyclic GMP Cytosol DNA DNA Binding DNA Repair Pathway DNA-Binding Proteins Data Data Analyses Dedications Defect Electron Microscopy Endosomes Enzyme-Linked Immunosorbent Assay Exposure to Fluorescence Microscopy Gene Expression Genes Genetic Genomic DNA Genomic Instability Goals Home Immune signaling Immunofluorescence Immunologic Immunotherapeutic agent Individual Innate Immune Response Interferons Interphase Intervention Learning Malignant Neoplasms Measures Membrane Proteins Mitosis Modeling Molecular Nuclear Nuclear Envelope Nuclear Inner Membrane Outcome Pathway interactions Phosphorylation Production Productivity Proteins Publishing Regulation Role Rupture Scientist Signal Transduction Site Sorting Source Stimulator of Interferon Genes System Testis Training Transcript Transfection Tumor Promotion Western Blotting Work cancer cell career chromosome missegregation design env Gene Products experimental study innate immune pathways insight light microscopy live cell imaging micronucleus novel recruit response seal sensor spatiotemporal synergism tumor tumor growth

项目摘要

PROJECT SUMMARY/ABSTRACT: In cancer cells, exposure of self DNA to the cytosol is driven by a variety of genomic instabilities such as micronuclei, chromatin bridges, and nuclear ruptures. This cytosolic DNA can be recognized by cytosolic DNA sensors such as cGAS (cyclic GMP-AMP synthase), which triggers a downstream innate immune response. Interestingly and confoundingly, the activation of the cGAS/STING innate immune pathway can protect or sensitize tumors to immunotherapeutic interventions depending on the specific context. Therefore, insight into the ways in which cGAS/STING signaling is regulated in cancer can inform targeted intervention. Sources of cytosolic DNA in cancer cells arise primarily from defects in mitosis that lead to the enclosure of chromosomes in micronuclei that are prone to rupture. These ruptured micronuclei recruit cGAS and nuclear envelope reformation (NER) factors—such as LEM2, CHMP7, and BAF—but it remains unknown how, or if, these NER factors impact cGAS/STING signaling but there is emerging evidence in published and in our preliminary data that there is potential crosstalk between cGAS/STING signaling and NER proteins. The goal of this proposal is to provide key insights into the regulation of the innate immune response to cytosolic DNA in cancer cells by nuclear envelope reformation factors. In order to achieve this goal, I will use transfected herring testes (HT) DNA and transfected DNA-coated beads as models for cytosolic DNA as this can be more readily controlled compared to the stochastic formation of micronuclei, only some of which are unstable and prone to rupture. With this model, I will use CRISPR/Cas9 gene-editing and the auxin-inducible-degron (AID) conditional degradation system to probe the roles of NER factors in cGAS/STING signaling in response to transfected HT DNA and DNA beads. This proposal will address fundamental aspects of cell biology and innate immune signaling that will shed light on immunotherapeutic targets for cancer.

项目概要/摘要：在癌细胞中，自身 DNA 暴露于细胞质是由多种基因组不稳定性驱动的，例如微核、染色质桥和核断裂可以被胞质 DNA 识别。 cGAS（环化 GMP-AMP 合酶）等传感器，可触发下游先天免疫反应。令人困惑的是，cGAS/STING 先天免疫途径的激活可以保护或根据具体情况使肿瘤对免疫治疗干预敏感。因此，需要深入了解。 cGAS/STING 信号在癌症中的调节方式可以为靶向干预提供信息。癌细胞中的胞质 DNA 主要源自有丝分裂缺陷，导致染色体封闭这些破裂的微核会招募 cGAS 和核膜。重组 (NER) 因子，例如 LEM2、CHMP7 和 BAF，但仍不清楚这些 NER 如何或是否会影响 cGAS/STING 信号传导的因素，但已发表的数据和我们的初步数据中有新的证据 cGAS/STING 信号和 NER 蛋白之间存在潜在的串扰该提案的目标。旨在为癌症中细胞质 DNA 的先天免疫反应的调节提供重要见解为了实现这一目标，我将使用转染的鲱鱼。测试 (HT) DNA 和转染的 DNA 包被珠作为胞质 DNA 的模型，因为这更容易与微核的随机形成相比，微核是受控的，只有其中一些是不稳定的并且容易发生在这个模型中，我将使用 CRISPR/Cas9 基因编辑和生长素诱导降解决定子 (AID) 条件。降解系统探测 NER 因子在 cGAS/STING 信号传导中对转染 HT 的反应的作用 DNA 和 DNA 珠子将解决细胞生物学和先天免疫的基本问题。信号将揭示癌症的免疫治疗靶点。