Deconstructing and targeting aneuploidy in human cancer - Resubmission - 1

摘要 非整倍体--染色体获得和丢失的存在--在正常组织中非常罕见，但在更多的 超过80%的人类肿瘤，特别是实体肿瘤。肿瘤中高水平的非整倍体与较高的 疾病分级、肿瘤进展和对治疗的抵抗。非整倍体是否以及如何有助于 人类肿瘤的形成和发展还不是很清楚。鉴于基因组和临床研究在 癌症患者认为非整倍体驱动肿瘤发生，小鼠模型和体外实验研究 到目前为止，系统对非整倍体在肿瘤中的作用产生了相互矛盾的数据。人类肿瘤通常是 与特定染色体数目的增加或减少特别相关(S)。其中一个 取得进展的主要障碍是技术上的限制，即不能设计出“正确”类型的 “右”细胞类型中的非整倍体。 我们的最终目标是剖析非整倍体是否以及如何有助于 人类肿瘤。我们这里的第一个目标是生成能够真实概括非整倍体的细胞模型。 在肿瘤中发现的模式，以研究非整倍体如何影响肿瘤细胞的病理生物学(它们的能力 在体外或体内生长，逃避细胞死亡途径，在细胞压力下生存，调节转录和 译文)。我们的第二个目标是潜在地发现漏洞和合成致命的交互作用 与非整倍体状态相关。拟议项目的成果将为 实现我们实验室的长期目标，即更好地了解原因和 人类肿瘤中非整倍体的后果，以揭示非整倍体相关的生物标志物和 治疗靶点。 为了实现这一目标，我们将首先使用一组新生成的人类细胞，其中包含不同的 比较几种肿瘤相关细胞二倍体和非整倍体细胞的异倍体程度和类型 在体外和体内的表型。其次，我们将采用系统的方法来识别基因和 当被阻断时，抑制非整倍体肿瘤细胞的增殖和存活的通路，但不能抑制正常细胞。第三, 我们将主要对结直肠癌患者的样本进行蛋白质和磷酸蛋白分析，以进行解剖 非整倍体在蛋白质稳定性和途径调控水平上的后果。我们的结果将填补一个 在我们目前对非整倍体在肿瘤发生和发展过程中如何演变的理解中存在着重要的知识空白 我们如何利用这些知识来改善患者的预后。

Summary Aneuploidy—the presence of chromosome gains and losses—is very rare in normal tissues but occurs in more than 80% of human tumors, especially in solid tumors. A high level of aneuploidy in tumors correlates with higher- grade disease, tumor progression, and resistance to therapy. Whether and how aneuploidy contributes to formation and progression of human tumors is not well understood. Whereas genomic and clinical studies in cancer patients suggest that aneuploidy drives tumorigenesis, experimental studies in mouse models and in vitro systems has so far yielded conflicting data on the role of aneuploidy in tumors. Human tumors are often specifically associated with either increases or decreases in the number of specific chromosome(s). One of the main obstacles to progress has been the technical limitation of not being able to engineer the ‘right’ type of aneuploidy in the ‘right’ cell type. Our ultimate goal is to dissect whether and how aneuploidy contributes to initiation and progression or human tumors. Our first objective here is to generate cellular models that faithfully recapitulate the aneuploidy patterns found in tumors in order to study how aneuploidy affects the pathobiology of tumor cells (their ability to grow in vitro or in vivo, to evade cell death pathways, to survive cellular stress and to regulate transcription and translation). Our second objective is to uncover vulnerabilities and synthetic lethal interactions potentially associated with the aneuploid state. The outcomes of the proposed project will represent the foundation to achieve the long-term goal of our lab, which is to develop a better understanding of the causes and consequences of aneuploidy in human tumors in order to uncover aneuploidy-associated biomarkers and therapeutic targets. To accomplish this goal, we will first use a panel of newly generated human cells containing different degrees and types of aneuploidy to compare diploid and aneuploid cells for several tumor-related cellular phenotypes both in vitro and in vivo. Secondly, we will adopt a systematic approach to identify genes and pathways that when blocked, inhibit proliferation and survival of aneuploid tumor cells but not normal cells. Third, we will perform a protein and phospho-protein analysis mainly of colorectal tumor patients’ samples to dissect the consequences of aneuploidy at the level of protein stability and pathway regulation. Our results will fill an important gap of knowledge in our current understanding of how aneuploidy evolves during tumorigenesis and how we might take advantage of this knowledge to improve patient outcomes.