Leveraging cancer-specific defects in nuclear integrity to inform novel synthetic lethal strategies

利用癌症特异性的核完整性缺陷为新型合成致死策略提供信息

基本信息

批准号：
9886210
负责人：
MEGAN C KING
金额：
$ 18.22万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9886210
关键词：
Animal Model Appearance Architecture CRISPR interference CRISPR screen Cancer cell line Cell Death Cell Line Cell Nucleus Cell Survival Cell division Cell physiology Cells Cellular biology Cessation of life Chemicals Chromosomes Clustered Regularly Interspaced Short Palindromic Repeats DNA Defect Development Diagnosis Diagnostic Neoplasm Staging Dropout Drug Design Drug Screening Essential Genes Etiology Exposure to Future Genes Genetic Genetic Predisposition to Disease Goals HCT116 Cells Human Cell Line Immune checkpoint inhibitor Immune response Immune system Immunotherapy Individual Innate Immune Response Interphase Lamin Type A Lead Link Malignant Neoplasms Mechanics Membrane Modeling Morphology Mutation Normal Cell Nuclear Nuclear Envelope Nuclear Lamin Oncogenic Ontology Pathway interactions Proteins Rupture Shapes Signal Pathway Site Source Stimulator of Interferon Genes Structure System Systems Biology The Cancer Genome Atlas Tissues Tumor Suppressor Proteins base cancer cell cancer diagnosis cancer genetics cell transformation cell type combat effective therapy experimental study fitness gene product genome-wide genomic data healing immune clearance inhibitor/antagonist innate immune pathways insight metaplastic cell transformation neoplastic cell new therapeutic target novel novel therapeutics personalized medicine potential biomarker recruit repaired response screening sensor support network tumor tumorigenesis

项目摘要

Summary Altered nuclear shape and appearance has long been known to be pathognomonic for cellular transformation; as a consequence, it is a critical parameter used in cancer diagnosis and tumor grading. Despite an increasingly mechanistic understanding of oncogenic and tumor suppressor pathways, as well as burgeoning genomic data that heralds the possibility of personalized treatments, we still lack a firm understanding of the relationship between nuclear architecture and cancer. In particular, it has yet to be defined if changes in the nucleus are causal or simply a consequence of transformation. Here, we sidestep this question, and instead ask: can the changes in nuclear architecture typical of cancer cells be exploited as a liability? Altered nuclear shape is intimately tied to mechanical defects of the nuclear envelope; recently, such defects have been linked to either transient or catastrophic losses of nuclear integrity, which can lead to cell death through two potential mechanisms. First, permanent losses of nuclear integrity are incompatible with cellular viability. Second, even transient losses of the nuclear barrier expose the DNA to cytoplasmic DNA sensors such as cGAS, which can drive a STING-dependent innate immune response that, at least in some cases, is sufficient to drive cell- autonomous death. In the latter case, loss of nuclear integrity also boosts the immune response to the tumor. Importantly, pathways that recognize and “heal” ruptures of the nuclear envelope have also been recently defined; perhaps not surprisingly, these repair mechanisms become critical for cell viability in contexts where nuclear integrity is compromised. Taken together, these new insights make a strong case that further weakening nuclear integrity in tumor cells can be exploited to drive cell death and immune system recognition. Here, in Aim 1, we propose to leverage an unbiased, genome-wide CRISPR dropout screen to identify synthetic lethal interactions of 1) normal cells with either weakened nuclear integrity or defective nuclear repair mechanisms or 2) cancer cell lines, with and without further compromise of their nuclear integrity pathways. In Aim 2, we will apply systems level approaches to organize the resulting context-dependent fitness genes into functional nodes. Beyond the strength of the genetic interaction, targets for in depth analysis will be further prioritized based on the availability of chemical inhibitors and representation in The Cancer Genome Atlas. Mechanistic experiments will explicitly examine these high priority synthetic genetic relationships in the context of nuclear shape, nuclear ruptures, and innate immune pathway activation. Completion of these two Aims will lead to the development of novel targets that exploit a key pathognomonic structure for cancer.

概括长期以来，人们都知道核形状和外观的改变是细胞转化的特征。因此，它是癌症诊断和肿瘤分级中使用的关键参数。对致癌和肿瘤抑制途径的机械性理解日益加深，以及新兴的基因组数据预示着个性化治疗的可能性，但我们仍然缺乏对基因组数据的深刻理解特别是，核结构与癌症之间的关系尚未确定。核心是因果关系，或者仅仅是转化的结果。在这里，我们回避这个问题，而是回避这个问题。问：癌细胞典型的核结构变化是否可以被利用为一种责任？形状与核膜的机械缺陷密切相关，最近，此类缺陷已被联系起来；核完整性的短暂或灾难性丧失，这可能通过两种潜在的方式导致细胞死亡首先，核完整性的永久丧失与细胞活力是不相容的。核屏障的短暂丧失将 DNA 暴露于细胞质 DNA 传感器，例如 cGAS，这可以驱动依赖于 STING 的先天免疫反应，至少在某些情况下，足以驱动细胞- 在后一种情况下，核完整性的丧失也会增强对肿瘤的免疫反应。重要的是，最近也发现了识别和“治愈”核膜破裂的途径定义；也许并不奇怪，这些修复机制在以下情况下对细胞活力至关重要。总而言之，这些新见解进一步证明了核完整性受到损害。肿瘤细胞核完整性的减弱可用于驱动细胞死亡和免疫系统识别。在此，在目标 1 中，我们建议利用公正的全基因组 CRISPR 缺失筛选来识别 1) 核完整性减弱或核修复缺陷的正常细胞的合成致死相互作用机制或2）癌细胞系，无论是否进一步损害其核完整性途径。目标 2，我们将应用系统级方法将所得的上下文相关的适应基因组织成除了基因相互作用的强度之外，深入分析的目标还将进一步确定。根据化学抑制剂的可用性和癌症基因组图谱中的代表性确定优先顺序。机制实验将明确地检查背景中这些高优先级的合成遗传关系核形状、核破裂和先天免疫途径激活的完成将实现这两个目标。导致开发利用癌症关键特征结构的新靶点。