Tracking the evolution of breast cancer through single cell analyses of premalignant breast tissues from women at high risk for cancer development

通过对癌症高危女性的癌前乳腺组织进行单细胞分析来追踪乳腺癌的演变

• 批准号: 10249258
• 负责人: Joan Siefert Brugge
• 金额: $ 85.16万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10249258
• 关键词: BRCA mutations; BRCA1 Mutation; BRCA1 gene; BRCA2 Mutation; BRCA2 gene; Breast; Breast Cancer Prevention; Breast Cancer Risk Factor; Cell Lineage; Cell Proliferation; Cells; Cytometry; DNA Damage; Development; Evolution; Goals; High Risk Woman; High-Risk Cancer; Histology; Human; Image; Immunocompromised Host; Immunofluorescence Immunologic; In Situ; Individual; Inherited; Intervention; Malignant Neoplasms; Mammary Gland Parenchyma; Mammary Neoplasms; Masks; Mus; Mutation; Organoids; Periodicity; Population; Population Analysis; Premalignant Cell; Premalignant Change; Proliferation Marker; Quality of life; RNA; Structure; Surface; Susceptibility Gene; Technology; Tissues; Woman; brca gene; high risk; malignant breast neoplasm; mammary; mutant; mutation carrier; novel diagnostics; premalignant; prevent; programs; prophylactic; prophylactic mastectomy; psychologic; reconstitution; single cell analysis; single cell technology; single-cell RNA sequencing; tool; tumor; tumor progression; tumorigenesis

Abstract: The overarching objective of the proposed studies is to identify and characterize early premalignant changes in breast tissues from women that carry genetic alterations associated with a high risk of breast cancer to ultimately develop strategies to detect and prevent the development of breast cancer. To accomplish this goal, we have optimized three technologies to profile single breast mammary cells (MECs): (1) CyTOF mass cytometry to allow tracking in parallel of >30 cell lineage and proliferation markers, (2) single cell RNA sequencing to identify expression programs of cell populations enriched in mutation-carriers, and (3) multi-plex cyclic immunofluorescence imaging (CyCIF) to simultaneous image >50 markers in situ. These technologies make it possible to detect differences in small populations of cells that would be masked by bulk population analyses. To date, we have profiled breast tissues from over 30 women with wild-type or mutant BRCA1 or BRCA2 by CyTOF and have identified distinct, previously unrecognized subpopulations of cells that are enriched in breast tissues from BRCA1 and/or BRCA2 carriers. These enriched subpopulations may represent cells that are either directly on the path to malignancy or indirectly contribute to the development of cancer in these high-risk women. We have identified RNA signatures associated with these enriched subpopulations, which include surface markers to isolate them from breast tissue to investigate both these possibilities and to track them within breast tumors. The signatures associated with one of the enriched populations have provided clues as to the basis for their accumulation, as well as potential strategies to prevent their accumulation. Using CyCIF, we have been able to identify enriched subpopulations of cells in situ within breast tissues and track their association with aberrant histologies. We have also developed organoid cultures that maintain all of the major MEC lineages as well as the BRCA1/2-enriched populations, and that are able to reconstitute glandular structures in immunocompromised mice. We believe that these tools provide an unprecedented opportunity to track the development of human cancer. In the proposed studies, we will investigate whether and how the BRCA1/2+/mut-enriched subpopulations contribute to tumorigenesis in mutation carriers. We will also investigate the basis for the enrichment of these populations and the contribution of DNA damage to their enrichment. Later stage studies will focus on the development of strategies to interfere with tumor progression, and importantly to develop novel diagnostic strategies to inform on the timing of prophylactic interventions. In addition, we will examine tissues from women who carry mutations in other breast cancer predisposition genes to establish whether similar subpopulations are detected in other high-risk individuals. And lastly, we will examine the possibility that these cells represent cells-of-origin of sporadic breast tumors that arise more broadly in the population.

抽象的： 拟议研究的总体目标是识别和表征早期癌前变化 来自女性的乳腺组织携带与乳腺癌高风险相关的基因改变，最终 制定检测和预防乳腺癌发展的策略。为了实现这个目标，我们有 优化了三种技术来分析单个乳房乳腺细胞 (MEC)：(1) CyTOF 质谱流式分析仪，以允许 并行追踪 >30 个细胞谱系和增殖标记，(2) 单细胞 RNA 测序以识别 富含突变载体的细胞群的表达程序，以及（3）多重循环免疫荧光 成像 (CyCIF) 可同时对 > 50 个标记进行原位成像。这些技术使检测成为可能 小群体细胞中的差异可能会被大群体分析所掩盖。迄今为止，我们已经 通过 CyTOF 对 30 多名携带野生型或突变型 BRCA1 或 BRCA2 女性的乳腺组织进行了分析，并发现 鉴定出不同的、以前未被识别的细胞亚群，这些细胞亚群富含于乳腺组织中的 BRCA1 和/或 BRCA2 携带者。这些富集的亚群可能代表直接处于通往 恶性肿瘤或间接导致这些高危女性患癌症。我们已经鉴定出RNA 与这些丰富的亚群相关的特征，其中包括将它们与 乳腺组织来研究这两种可能性并在乳腺肿瘤内追踪它们。签名 与其中一个富集种群相关的物质也提供了关于其积累基础的线索，以及 作为防止其积累的潜在策略。使用 CyCIF，我们已经能够识别富集 乳腺组织内原位细胞亚群并追踪它们与异常组织学的关联。我们有 还开发了维持所有主要 MEC 谱系以及富含 BRCA1/2 的类器官培养物 群体，并且能够在免疫功能低下的小鼠中重建腺体结构。我们相信 这些工具为追踪人类癌症的发展提供了前所未有的机会。在提议的 研究中，我们将研究 BRCA1/2+/mut 富集亚群是否以及如何促进 突变携带者的肿瘤发生。我们还将调查这些人群富集的基础， DNA 损伤对其富集的贡献。后期研究将侧重于战略的制定 干扰肿瘤进展，重要的是开发新的诊断策略来告知治疗的时机 预防性干预措施。此外，我们还将检查其他乳房携带突变的女性的组织。 癌症易感基因以确定是否在其他高风险个体中检测到类似的亚群。 最后，我们将检查这些细胞代表散发性乳腺肿瘤的起源细胞的可能性 在人群中更广泛地出现。