Investigating the function of subclonal cooperation in breast cancer progression

研究亚克隆合作在乳腺癌进展中的功能

• 批准号: 10314659
• 负责人: Rachel Marie Nakagawa
• 金额: $ 3.77万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2025-03-25
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10314659
• 关键词: Investigating; function; subclonal; cooperation; breast

Project Summary/Abstract Intratumor genetic heterogeneity is present in a vast majority of solid cancers and correlates with tumor aggressiveness and poor therapeutic response. Whether tumor heterogeneity promotes tumor growth and survival, and the potential mechanisms driving these phenotypic changes, are open areas of investigation. To explore this fundamental gap in knowledge, I generated a model for studying subclonal cooperation and tumor heterogeneity in vitro and in vivo, respectively, using isogenic MCF10A cell lines that either overexpress MYC (MYC) or oncogenic HRASG12V (HRAS). MYC and oncogenic RAS are two canonical oncogenes that are known to signal in distinct pathways, though studies focused on MYC and HRAS interactions have primarily explored cell intrinsic mechanisms of cooperation. Therefore, I will use MYC and HRAS as an exemplar for subclonal cooperation to study their cooperativity in distinct cell populations. In preliminary studies using the MYC and HRAS model, I made the exciting observation that individual subclones expressing MYC or HRAS cooperate non-cell autonomously: cell proliferation increases when subclones are grown together in vitro and tumor growth is more rapid when subclones are co-injected into mice. Importantly, these data suggest targeting clonal interdependence may be necessary to improve therapeutic strategies. We hypothesize tumor subclones cooperate to elicit emergent tumor properties promoting tumor growth, and, potentially, tumor metastasis and drug resistance. The three Experimental Aims of this proposed research will rigorously test my hypothesis, studying the phenomenon of subclonal cooperation in cell culture and in orthotopic and patient derived xenograft (PDX) models. Experimental Aim 1 will test if subclonal cooperation is necessary for more rapid tumor initiation and proliferation, by deconstructing heterogenous tumors at distinct timepoints during tumor progression. Experimental Aim 2 will explore how tumor heterogeneity is functional by defining how exosome mediated protein transfer contributes to subclonal cooperation. Finally, in Experimental Aim 3, I will test whether subclonal interactions promote therapeutic resistance in orthotopic mouse mammary tumors generated using human- derived cell lines. Additionally, I will validate our findings of subclonal heterogeneity by characterizing the subclonal expression of specific oncogenes present in in a library of readily available PDX models. Together, these data will improve our understanding of subclonal cooperation in tumor progression, providing key insights for future work aimed at efficiently deconstructing and treating complex, heterogenous tumors.

项目摘要/摘要 肿瘤内遗传异质性存在于绝大多数固体癌症中，并且与肿瘤相关 侵略性和治疗反应不佳。肿瘤异质性是否促进肿瘤的生长和 生存以及推动这些表型变化的潜在机制是开放研究领域。到 探索知识的基本差距，我产生了一个用于研究亚克隆合作和肿瘤的模型 分别使用异基因MCF10A细胞系在体外和体内的异质性，要么过表达MYC （MYC）或致癌HRASG12V（HRAS）。 Myc和Oncentic Ras是两个已知的典型癌基因 在不同的途径中发出信号，尽管关注MYC和HRAS相互作用的研究主要探索了 细胞固有的合作机制。因此，我将使用MYC和HRA作为亚克隆的典范 合作研究其在不同细胞群体中的合作性。在使用MYC和 HRAS模型，我进行了令人兴奋的观察，即单个表达MYC或HRAS合作的单个子克隆 非细胞自主：当亚克隆在体外和肿瘤生长中一起生长时，细胞增殖会增加 将亚克隆共同注射到小鼠中时会更快。重要的是，这些数据表明针对克隆 相互依存可能是改善治疗策略所必需的。我们假设肿瘤亚克隆 合作以引起促进肿瘤生长的新兴肿瘤特性，并可能是肿瘤转移和 耐药性。 这项拟议的研究的三个实验目标将严格检验我的假设，研究 细胞培养以及原位和患者衍生异种移植物（PDX）中克隆合作的现象 型号。实验目标1将测试是否需要更快的肿瘤开始和 增殖，通过在肿瘤进展过程中在不同的时间点上解构异质肿瘤。 实验AIM 2将通过定义外泌体介导的蛋白的方式探索肿瘤异质性如何起作用 转移有助于亚克隆合作。最后，在实验目标3中，我将测试下克隆 相互作用促进使用人类产生的原位小鼠乳腺肿瘤的治疗性抗性 派生的细胞系。此外，我将通过表征来验证我们的亚克隆异质性的发现 在可用的PDX模型库中存在的特定癌基因的亚克隆表达。一起， 这些数据将提高我们对肿瘤进展中克隆合作的理解，提供关键的见解 用于未来的工作，旨在有效地解构和治疗复合物异质肿瘤。