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: 10459285
• 负责人: Rachel Marie Nakagawa
• 金额: $ 3.94万
• 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/10459285
• 关键词: Acceleration; Adopted; Area; Automobile Driving; Biochemical; Breast Cancer Model; Breast Epithelial Cells; Cell Culture Techniques; Cell Line; Cell Proliferation; Cell secretion; Cells; Cessation of life; Coculture Techniques; Collection; Communication; Complex; Cultured Cells; Data; Dependence; Development; Disease; Drug resistance; Engineering; Evolution; Future; Genetic Heterogeneity; Genetic Transcription; Goals; Heterogeneity; Human; In Vitro; Individual; Investigation; Knowledge; Label; Libraries; MCF10A cells; Maintenance; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Mediating; Modeling; Mus; Mutation; Neoplasm Metastasis; Oncogenes; Oncogenic; Pathway interactions; Patients; Phenotype; Population; Prognosis; Proliferating; Property; Proteins; Research; Signal Transduction; Solid; Solid Neoplasm; Sorting; Stains; Study models; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Tissues; Tumor Promotion; Tumor-Derived; Work; breast cancer progression; exosome; experimental study; human disease; improved; in vivo; insight; intercellular communication; neoplastic cell; overexpression; patient derived xenograft model; response; subclonal heterogeneity; suicide gene; targeted treatment; therapy resistant; transcriptome; transcriptome sequencing; treatment response; tumor; tumor behavior; tumor growth; tumor heterogeneity; tumor initiation; tumor progression; tumor xenograft; tumorigenic

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.

项目概要/摘要 肿瘤内遗传异质性存在于绝大多数实体癌中，并与肿瘤相关 攻击性和治疗反应差。肿瘤异质性是否促进肿瘤生长 生存以及驱动这些表型变化的潜在机制是研究的开放领域。到 为了探索这一知识上的根本差距，我生成了一个用于研究亚克隆合作和肿瘤的模型 使用过表达 MYC 的同基因 MCF10A 细胞系分别在体外和体内观察异质性 (MYC) 或致癌 HRASG12V (HRAS)。 MYC 和致癌 RAS 是已知的两种典型致癌基因 尽管针对 MYC 和 HRAS 相互作用的研究主要探索了不同途径中的信号传导 细胞内在的合作机制。因此，我将使用 MYC 和 HRAS 作为亚克隆的范例 合作研究它们在不同细胞群中的协同性。在使用 MYC 和 HRAS 模型，我做出了令人兴奋的观察，表达 MYC 或 HRAS 的个体亚克隆相互协作 非细胞自主：当亚克隆在体外一起生长和肿瘤生长时，细胞增殖增加 当将亚克隆共同注射到小鼠体内时，速度更快。重要的是，这些数据表明针对克隆 相互依存对于改善治疗策略可能是必要的。我们假设肿瘤亚克隆 合作引发促进肿瘤生长的新兴肿瘤特性，并可能促进肿瘤转移和 耐药性。 这项拟议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来源的异种移植物（PDX）中的亚克隆合作现象 模型。实验目标 1 将测试亚克隆合作是否对于更快速的肿瘤发生和 通过在肿瘤进展过程中的不同时间点解构异质肿瘤来促进增殖。 实验目标 2 将通过定义外泌体介导蛋白质的方式来探索肿瘤异质性如何发挥作用 转移有助于亚克隆合作。最后，在实验目标3中，我将测试是否亚克隆 相互作用促进使用人源产生的原位小鼠乳腺肿瘤的治疗耐药性 衍生细胞系。此外，我将通过表征亚克隆异质性来验证我们的发现 存在于现成的 PDX 模型库中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亚克隆合作的理解，提供关键见解 未来的工作旨在有效地解构和治疗复杂的异质肿瘤。