Transcriptional regulation of mammary gland development

乳腺发育的转录调控

• 批准号: 10364226
• 负责人: Eran Robert Andrechek
• 金额: $ 51.33万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10364226
• 关键词: Address; Adult; Alveolar; Biology; Breast Epithelial Cells; Cell Cycle; Cell Cycle Regulation; Cells; ChIP-seq; Clustered Regularly Interspaced Short Palindromic Repeats; Color; Data; Data Set; Databases; Defect; Development; Developmental Gene; Duct (organ) structure; E2F transcription factors; Embryo; Ensure; Excision; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Gland; Goals; Growth and Development function; Hydrocephalus; Image; In Vitro; Knock-out; Knockout Mice; Laboratories; Lactation; Lead; Malignant Neoplasms; Mammary gland; Mediating; Mission; Mus; Outcome; Pathway interactions; Pattern; Phase; Phenotype; Play; Pregnancy; Process; Puberty; Public Health; Publishing; Regulation; Reporter; Repression; Research; Role; Signal Pathway; Tamoxifen; Testing; Tissues; Transcription Coactivator; Transcription Repressor; Transcriptional Regulation; United States National Institutes of Health; Validation; Weaning; Work; base; cell type; conditional knockout; gene repression; in vivo evaluation; innovation; malignant breast neoplasm; mammary; mammary epithelium; mammary gland development; offspring; programs; reconstitution; single cell analysis; single-cell RNA sequencing

Mammary gland development and function requires precise stage and spatial control of transcriptional programs. These processes are tightly controlled to ensure the successful function of the mammary gland, including lactation which is essential for rearing of offspring. Recent work has demonstrated a role for the E2F transcription factors that goes well beyond their traditional role in cell cycle regulation. The role of the activator E2Fs (E2F1-3a) is well characterized, including regulation of mammary gland development. However, little is known of the role of the repressor E2Fs, E2F4 and E2F5, in the mammary gland and this is partially due to hydrocephaly and early lethality in the knockout strains. The long term goal of our work is to define the role of transcriptional regulation in mammary gland development and function. Understanding normal biology then informs our studies of how this goes awry and results in breast cancer. The immediate objective of this proposal, which is the next step in our long-term goal, is to precisely define the role of repressor E2Fs in the mammary gland. Our central hypothesis is that the repressor E2Fs regulate key mammary gland developmental genes. This hypothesis was based on preliminary data from knockout mice. Indeed, loss of E2F5 in the mammary epithelium resulted in delayed ductal extension during puberty, delayed involution and alveolar overgrowth in virgin adult mice. Loss of E2F4 also resulted in delayed ductal outgrowth but was also associated with a profound lack of alveolar expansion during pregnancy and lactation. Combining ChIP-Seq data and our gene expression data, we predicted both unique and shared E2F target genes with mammary development roles. The rationale for the proposed work is that once we have completed this proposal, we will understand how transcriptional repression regulates development of the mammary gland. We plan to test our central hypothesis and accomplish the objective of this application by investigating the following specific aims. In the first aim we will characterize the mammary gland phenotypes associated with the mammary epithelial cell specific knockout of E2F5. In the second aim we will generate ChIP-Seq data and integrate it with E2F5 induced gene expression data to determine E2F5 targets. Target genes will be filtered and prioritized using additional datasets describing mammary development. Finally, in the third aim we will examine scRNAseq data in virgin glands lacking E2F3 or E2F5. This proposal is innovative because it will elucidate the genetic mechanisms regulating mammary gland development and function by the repressor E2F transcription factors. This contribution is significant because it will establish a role for E2F5 in mammary gland development.

乳腺的发育和功能需要精确的阶段和空间控制， 转录程序。这些过程都受到严格控制，以确保成功的功能 哺乳期包括哺乳期，哺乳期是养育后代的必要条件。最近的工作 已经证明了E2 F转录因子的作用远远超出了传统的转录因子。 在细胞周期调控中的作用。激活剂E2 Fs（E2 F1 -3a）的作用已得到充分表征， 包括调节乳腺发育。然而，很少有人知道的作用， 阻遏物E2 Fs，E2 F4和E2 F5，在乳腺，这是部分由于脑积水 以及敲除菌株的早期致死性。我们工作的长期目标是确定 乳腺发育和功能的转录调控。理解正常 然后，生物学告诉我们这是如何出错并导致乳腺癌的研究。的 这一建议的近期目标，也是我们长期目标的下一步，是准确地 定义阻遏物E2 Fs在乳腺中的作用。我们的中心假设是， 阻遏物E2 Fs调节关键的乳腺发育基因。这个假设是基于 基因敲除小鼠的初步数据。事实上，乳腺上皮中E2 F5的缺失导致了 青春期乳腺导管延长延迟，处女患者退化延迟和肺泡过度生长 成年老鼠E2 F4的缺失也会导致导管生长延迟，但也与 在怀孕和哺乳期间肺泡极度缺乏扩张。组合ChIP-Seq数据 和我们的基因表达数据，我们预测了独特的和共享的E2 F靶基因， 乳腺发育的作用。拟议工作的理由是，一旦我们有了 完成这个建议，我们将了解转录抑制如何调节 乳腺的发育。我们计划测试我们的中心假设，并实现 本申请的目的是通过研究以下具体目标。在第一个目标中，我们将 表征与乳腺上皮细胞特异性 敲除E2 F5。在第二个目标中，我们将生成ChIP-Seq数据并将其与E2 F5整合 诱导基因表达数据以确定E2 F5靶标。目标基因将被过滤， 使用描述乳房发育的其他数据集进行优先排序。最后，在第三个目标中， 我们将检查缺乏E2 F3或E2 F5的处女腺中的scRNAseq数据。这一建议具有创新性 因为它将阐明调节乳腺发育的遗传机制， 由阻遏物E2 F转录因子起作用。这一贡献意义重大，因为它将 确定E2 F5在乳腺发育中作用。