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: 9816264
• 负责人: Joan Siefert Brugge
• 金额: $ 101.7万
• 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/9816264
• 关键词: 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; Mammary gland; Masks; Mus; Mutation; Organoids; Periodicity; Population; Population Analysis; Premalignant; Premalignant Change; Proliferation Marker; Quality of life; RNA; Structure; Surface; Susceptibility Gene; Technology; Tissues; Woman; cancer cell; high risk; malignant breast neoplasm; mutant; mutation carrier; novel diagnostics; 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： 拟议的研究的总体目标是识别和表征早期的早期预言变化 来自女性的乳房组织，这些乳腺组织具有与乳腺癌高风险相关的遗传改变，以最终 制定策略来检测和防止乳腺癌的发展。为了实现这一目标，我们有 优化了三种技术以介绍单乳腺乳腺细胞（MECS）：（1）质量细胞仪以允许 并行跟踪> 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体的表达程序和（3）多质子循环免疫荧光 成像（cycif）与同时图像> 50标记原位。这些技术使检测到可能 大量种群分析将掩盖的小细胞种群的差异。迄今为止，我们有 由30多名女性具有野生型或突变体BRCA1或BRCA2的乳房组织，并具有 确定了富含BRCA1乳房组织的细胞的不同的，以前未被认可的细胞亚群 和/或BRCA2载体。这些富集的亚群可以代表直接在通往路径上的细胞 这些高危女性的恶性肿瘤或间接有助于癌症的发展。我们已经确定了RNA 与这些丰富的亚群相关的签名，其中包括表面标记以将其隔离 乳房组织以研究这两种可能性并在乳腺肿瘤中进行跟踪。签名 与一个丰富人口之一相关的人也提供了有关其积累基础的线索 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 细胞在乳房组织中原位的亚群，并跟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BRCA1/2富集的器官培养物 种群，能够在免疫功能低下的小鼠中重新建立腺结构。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。在提议中 研究，我们将研究BRCA1/2+/MUT元素的亚群是否有助于 突变载体中的肿瘤发生。我们还将调查这些人群丰富的基础和 DNA损伤对其富集的贡献。后来的研究将重点放在战略的制定上 干扰肿瘤进展，重要的是开发新颖的诊断策略以告知 预防性干预措施。此外，我们将检查来自其他乳房中携带突变的女性的组织 癌症易感基因确定在其他高危个体中是否检测到类似的亚群。 最后，我们将研究这些细胞代表偶发性乳腺肿瘤的细胞细胞的可能性 人口更广泛地出现。