Dissecting the impact of tumor-intrinsic chromosomal instability on the cancer ecosystem

项目总结 这项提议的目标是发现癌细胞中正在进行的染色体错误分离事件(a 称为染色体不稳定的过程，或CIN)改变肿瘤生态系统，促进癌症进展。 长期以来，染色体拷贝数改变(也称为非整倍体)与免疫相关。 抑制表型、耐药和转移。然而，除了非整倍体之外，它仍然是未知的 正在进行的染色体错误分离过程是否会导致肿瘤进展。利用前女友- 周边工具，使我们能够拨号向上或向下拨号染色体错误分离率在其他相同的基因 背景：我们先前已经发现，正在进行的染色体错误分离会导致容易破裂的错配。 核仁，使基因组双链DNA(DsDNA)暴露在胞浆中，导致持久激活 CGAS刺痛的先天免疫途径(1)。然而，与其促进强健的I型干扰素(干扰素)重新- CIN激活癌细胞中的SUNSE、STING促进非典型核因子-kB(NC-NF-kB)信号转导 一种未知的机制--驱动转移。CIN驱动的转移在多大程度上取决于 免疫系统尚不清楚。我们做了一个令人惊讶的观察，CIN在一个 通过塑造癌细胞与肿瘤微环境之间的相互作用，实现癌细胞的非自主性。 环境(TME)。这一发现推动了一种全新的、系统级的方法的发展 评估TME中称为接触跟踪的细胞-细胞相互作用的性质和条件依赖性。这 方法利用单细胞RNA测序技术捕捉到的内在生物变异，推断细胞和细胞间的相互作用。 在事先不知道下游靶基因的情况下，对配体-受体介导的相互作用的LAR反应。 将这一创新的计算工具与CIN和STING的遗传扰动相结合，我们发现CIN 通过诱导癌细胞固有的内质网应激反应，产生促转移的TME。在目标1中，我们建议 从机制上剖析CIN、STING和内质网应激在肿瘤进展过程中的上位关系 用三阴性乳腺癌(TNBC)来确定ER应激是否能代表慢性乳腺癌的治疗靶点。 整合体不稳定的肿瘤。我们还将测试内质网应激反应是否是NC-NF-kB激活的基础。 在目标2下，我们将改进肿瘤衍生配体效应的因果推断，并探索其分子基础。 用基因调控网络询问CIN依赖的细胞-细胞相互作用网络是否保守 使用人类数据和乳腺癌、胰腺癌和肺癌的小鼠模型，研究各种癌症类型。这个 这些方法的融合，结合我们对CIN在癌症中的深刻理解，有望阐明-- CIN诱导的刺痛信号在疾病进展过程中对肿瘤免疫串扰的复杂作用。 重要的是，这项工作将揭示针对染色体不稳定肿瘤的新策略， 否则很难治疗。

PROJECT SUMMARY The goal of this proposal is to discover how ongoing chromosome missegregation events in cancer cells (a process called chromosomal instability, or CIN) alters the tumor ecosystem to promote cancer progression. Chromosome copy number alterations (also referred to as aneuploidy) have long been associated with immune suppressive phenotypes, drug resistance, and metastasis. Beyond aneuploidy, however, it remains unknown whether the ongoing process of chromosome missegregation gives rise to tumor progression. Harnessing ex- perimental tools that enable us to dial-up or dial-down chromosome missegregation rates in otherwise isogenic backgrounds, we have previously found that ongoing chromosome missegregation generates rupture-prone mi- cronuclei, which expose genomic double-stranded DNA (dsDNA) to the cytosol, leading to persistent activation of the cGAS-STING innate immune pathway (1). Yet, instead of promoting a robust type I interferon (IFN) re- sponse, STING activation in cancer cells with CIN promotes noncanonical NF-kB (nc-NF-kB) signaling – through an unknown mechanism – to drive metastasis. The extent to which CIN-driven metastasis is dependent on the immune system remains unknown. We made the surprising observation that CIN drives tumor progression in a cancer cell non-autonomous manner by shaping the interaction between cancer cells and the tumor microenvi- ronment (TME). This finding motivated the development of a fundamentally new, systems-level approach to evaluate the nature and conditional-dependence of cell-cell interactions in the TME called ContactTracing. This method exploits intrinsic biological variance captured by single cell RNA sequencing technologies, to infer cellu- lar responses to ligand-receptor mediated interactions without prior knowledge of downstream target genes. Combining this innovative computational tool with genetic perturbation of CIN and STING we found that CIN engenders a pro-metastatic TME by inducing a cancer cell-intrinsic ER-stress response. In Aim 1, we propose to mechanistically dissect the epistatic relationship between CIN, STING, and ER-stress in the progression of triple negative breast cancer (TNBC) to determine whether ER-stress can represent a therapeutic target in chro- mosomally unstable tumors. We will also test whether an ER-stress response underlies nc-NF-kB activation. Under Aim 2, we will improve causal inference of tumor-derived ligand effects and explore their molecular basis using gene regulatory networks to ask whether CIN-dependent cell-cell interaction networks are conserved across cancer types using both human data and mouse models of breast, pancreatic, and lung cancers. The amalgamation of these approaches, combined with our deep understanding of CIN in cancer, is poised to eluci- date the complex roles of CIN-induced STING signaling on tumor-immune crosstalk during disease progression. Importantly, this work is poised to reveal novel strategies aimed at targeting chromosomally unstable tumors, which are otherwise difficult to treat.