Deciphering the mechanism underlying BRCA1 breast cancer development

破译 BRCA1 乳腺癌发生的机制

基本信息

批准号：
10218120
负责人：
DAVID Morse LIVINGSTON
金额：
$ 105.34万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-07 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10218120
关键词：
Affect Animals BARD1 gene BRCA1 Mutation BRCA1 gene Binding Biochemical Biological Breast Breast Epithelial Cells Cells Clustered Regularly Interspaced Short Palindromic Repeats Complex DNA Damage DNA Repair Data Development Disease ES01 Event FANCD2 protein Family Frequencies Future Genes Genomics Individual Inherited Interruption Lead Life Malignant Neoplasms Malignant neoplasm of ovary Menarche Methods Molecular Mutate Numbness Operative Surgical Procedures Pathway interactions Physiological Precancerous Conditions Protein Overexpression Proteins Publishing Research Resort Role SMARCA4 gene Series Testing Translating Tumor Suppressor Proteins cell dedifferentiation experimental study loss of function malignant breast neoplasm mammary epithelium member mouse model novel strategies prevent single-cell RNA sequencing stem cell proliferation tumor

项目摘要

Project Summary/Abstract Inherited BRCA1 mutations very often result in high frequency breast and ovarian cancer in affected families. This proposal is aimed at illuminating key steps that occur after menarche in tumor-free, BRCA1-mutated mammary epithelium and contribute in a major way to future BRCA1 breast cancer (BrCa) development. Our published data and work in progress suggest the existence of a series of interrelated biochemical steps that, when interrupted, result in the conversion of ostensibly normal, BRCA1- mutated breast epithelial cells into an abnormal, dedifferentiated, unusually primitive, and likely pre-malignant state. This change involves the loss of function of any member of a group of 6 particular genes which translates into a breakdown in DNA damage control which, in turn, promotes a breakdown in mammary epithelial differentiation. In breast cells these genes normally communicate with one another, functionally, as if they were components of a biologically important pathway dedicated, in part, to BrCa suppression. Indeed, their protein products form a distinct molecular complex which also contains the BRCA1 tumor suppressor protein, p220, and this complex operates in ways that suggest a major role for it in such a pathway. Among the 6 other complex members (BRG1, FancD2, CtIP,NUMB, HES-1, and BARD1), some (e.g. FancD2, BRG1) are somatically mutated in sporadic BrCa. Another (Numb) is a known BrCa suppressor in animals and a supporter of normal stem cell proliferation, and at least 5 of the 6 must be present and bind to p220 for the entire complex to form and mammary epithelial differentiation to proceed normally. Our work in progress also suggests that the integrity of this complex is required to prevent BRCA1 BrCa from developing. The proposed research will test the validity of this hypothesis along with the physiological importance of such a pathway, using a combination of cutting edge methods. Included will be an effort to a) test for lineage relationships between failed mammary epithelial cell (MEC) DNA damage repair and MEC dedifferentiation and between MEC dedifferentiation and BRCA1 tumor development and b) to decipher the mechanisms governing the interrelationships of these events. A major component of these efforts will be the application of single cell RNAseq to assess the above-noted lineage relationships and mammary epithelium- focused genomic CRISPR and protein overexpression screens to detect individual proteins that participate in these important steps in BRCA1 BrCa development. We will use a newly developed mouse model in these experiments.!

项目摘要/摘要遗传的BRCA1突变通常会导致受影响家庭的高频乳腺癌和卵巢癌。该提案旨在阐明一级初经无肿瘤，brca1杂种后发生的关键步骤乳腺上皮，并为未来的BRCA1乳腺癌（BRCA）发育做出了重大贡献。我们已发布的数据和正在进行的工作表明存在一系列相互关联的生化步骤当中断时，这会导致表面上正常的BRCA1突变乳腺上皮细胞的转化变成异常，去分化，异常原始且可能的恶性状态。这种变化涉及一组6个特定基因的任何成员的功能丧失，这些基因转化为DNA中的分解损伤控制又促进了乳腺上皮分化的崩溃。在乳房细胞中这些基因通常在功能上相互通信，好像它们是生物学上的组成部分重要的途径专门用于BRCA抑制。确实，它们的蛋白质产品形成了独特的分子络合物还包含BRCA1肿瘤抑制蛋白P220，并且该复合物起作用在这种途径中暗示其主要作用的方式。在其他6个复杂成员（BRG1，FANCD2，CTIP，NUMB，HES-1和BARD1）中，有些（例如FANCD2， BRG1）在零星的BRCA中体外突变。另一个（麻木）是动物中已知的BRCA抑制器，正常干细胞增殖的支持者，并且至少有6个必须存在，并与P220结合。整个复合物形成和乳腺上皮分化以正常进行。我们正在进行的工作表明需要这种复合物的完整性来防止BRCA1 BRCA发展。拟议的研究将检验该假设的有效性，以及这种假设的重要性途径，结合尖端方法的组合。其中包括的努力是a）测试谱系失败的乳腺上皮细胞（MEC）DNA损伤修复与MEC去分化与在MEC去分化和BRCA1肿瘤的发展和b）中分解了控制机制这些事件的相互关系。这些努力的主要组成部分将是应用单细胞RNASEQ评估上述标题谱系关系和乳腺上皮浓缩基因组CRISPR和蛋白质过表达筛选检测参与BRCA1 BRCA开发中这些重要步骤的单个蛋白质。我们将使用在这些实验中，新开发的鼠标模型。