Transcriptional regulation of mammary gland development

乳腺发育的转录调控

• 批准号: 10649420
• 负责人: Eran Robert Andrechek
• 金额: $ 47.97万
• 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/10649420
• 关键词: Ablation; 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; Growth and Development function; Hydrocephalus; Image; In Vitro; Knock-out; Knockout Mice; Laboratories; Lactation; 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; Sorting; Tamoxifen; Testing; Tissues; Transcription Coactivator; Transcription Repressor; Transcriptional Regulation; United States National Institutes of Health; Validation; Weaning; Work; cell type; conditional knockout; gene repression; in vivo evaluation; inducible Cre; 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。这些过程受到严格控制，以确保成功的功能 乳腺，包括泌乳，这对于饲养后代至关重要。最近的工作 已经证明了E2F转录因子的作用，这远远超出了传统 在细胞周期调节中的作用。激活剂E2F（E2F1-3A）的作用很好， 包括调节乳腺发育。但是，知之甚少 在乳腺中，阻遏物E2FS，E2F4和E2F5，这部分是由于脑畸形 和淘汰赛中的早期致命性。我们工作的长期目标是定义 乳腺发育和功能中的转录调节。了解正常 然后，生物学会为我们的研究介绍如何出现问题并导致乳腺癌。这 该提案的直接目标，这是我们长期目标的下一步，就是确切 定义抑制剂E2F在乳腺中的作用。我们的核心假设是 阻遏物E2FS调节关键的乳腺发育基因。这个假设是基于 关于敲除小鼠的初步数据。实际上，乳腺上皮中E2F5的损失导致 在青春期期间的导管延伸延迟，处女中的延迟相关和肺泡过度生长 成年小鼠。 E2F4的损失也导致导管延迟出生，但也与 在怀孕和哺乳期间严重缺乏肺泡扩张。结合芯片序列数据 和我们的基因表达数据，我们预测了独特的和共享的E2F目标基因 乳腺发展角色。拟议工作的理由是，一旦我们拥有 完成了该建议，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完成 通过研究以下特定目的，该应用的目标。在第一个目标中，我们将 表征与乳腺上皮细胞特异性相关的乳腺表型 E2F5的敲除。在第二个目标中，我们将生成芯片序列数据并将其与E2F5集成 诱导基因表达数据确定E2F5靶标。靶基因将被过滤，并且 使用描述乳腺发展的其他数据集优先考虑。最后，第三个目标 我们将检查缺乏E2F3或E2F5的处女腺中的SCRNASEQ数据。该建议是创新的 因为它将阐明调节乳腺发育和 阻遏物E2F转录因子的功能。这项贡献很重要，因为它将 确立E2F5在乳腺发育中的作用。