Genomic and functional investigations of the transcriptional regulatory network of salivary gland morphogenesis and stem cell fate choices in defined genetic models

在确定的遗传模型中对唾液腺形态发生和干细胞命运选择的转录调控网络进行基因组和功能研究

• 批准号: 10554329
• 负责人: Rose-Anne Romano
• 金额: $ 37.74万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10554329
• 关键词: Address; Adult; Alleles; Animals; Area; Biological Markers; Biological Models; Biology; Cancer Patient; Cells; Clinical; Critical Pathways; Development; Disease; Ectodermal Dysplasia; Embryonic Development; Ensure; Epithelium; Equilibrium; Exhibits; Functional disorder; Gene Expression; Gene Expression Profile; Genetic; Genetic Models; Genetic Transcription; Genomics; Gland; Goals; Homeostasis; Human; Injury; Investigation; Knock-out; Knockout Mice; Knowledge; Maintenance; Mammary gland; Molecular; Molecular Profiling; Morphogenesis; Mus; Natural regeneration; Normal tissue morphology; Organ; Organogenesis; Outcome; Pathway interactions; Patients; Phenotype; Play; Population; Process; Proliferating; Protein Isoforms; Radiation Tolerance; Radiation induced damage; Regulator Genes; Reporting; Resolution; Role; Salivary Glands; Signal Pathway; Signal Transduction; Sjogren' s Syndrome; Skin; Specific qualifier value; Stratification; Submandibular gland; Systems Biology; Technology; Testing; Therapeutic; Time; Tissue-Specific Gene Expression; Tissues; Transgenic Mice; Xerostomia; conditional knockout; defined contribution; epithelial stem cell; experimental study; gamma irradiation; genetic approach; genomic tools; gland development; injured; insight; interest; irradiation; malignant mouth neoplasm; mouse model; novel; novel therapeutic intervention; programs; radiation-induced injury; regeneration following injury; regeneration function; regenerative; repaired; response; response to injury; self-renewal; single-cell RNA sequencing; stem; stem cell differentiation; stem cell division; stem cell fate; stem cell function; stem cell population; stem cells; tissue regeneration; transcription factor; transcription regulatory network; transcriptome sequencing; transcriptomics

PROJECT SUMMARY The intricate and dynamic balance between self-renewal, proliferation and differentiation of stem/progenitor cells of the salivary gland (SG) must be tightly regulated to ensure proper morphogenesis, homeostasis and regeneration. Alterations to normal SG function, clinically often manifested with hyposalivation, are associated with diseases such as Ectodermal Dysplasias, Sjögren’s Syndrome and with γ- irradiation therapy of oral cancer patients. Currently, treatment options for hyposalivation remain limited. Therefore, identifying crucial transcriptional and signaling networks that govern stem/progenitor cell function of the SG are much needed to facilitate targeted stem cell and regenerative-based therapies. ΔNp63, a stem cell enriched transcription factor, plays a critical role in epithelial regenerative function as evident from the phenotype of ΔNp63-null animals which exhibit developmental arrest and agenesis of epithelial-rich organs including the SG. However, our current knowledge of the molecular mechanisms by which ΔNp63 directs gene expression programs necessary for the commitment, maintenance and differentiation of the stem/progenitor cell population in the SG is lacking. Thus, identifying the p63-driven regulatory networks, particularly in the global and genomic context, is a key step towards a better understanding of the biology of SG stem/progenitor cells and ultimately in directing new strategies in treating SG dysfunction. To address these knowledge gaps, we will utilize multiple versatile mouse models to study two major independent areas of interest. First, we will use conditional knockout mouse models to examine the role of ΔNp63 in SG morphogenesis and in adult tissue maintenance and repair (Aim1). Such systematic studies are much needed as they will identify for the first time, the functional role of p63 in SG development and in orchestrating stem/progenitor cell differentiation programs. Second, we will use p63 knockout mouse models and lineage tracing experiments to determine the contribution of p63+ stem and progenitor cells during SG regeneration and in response to irradiation induced damage. Furthermore, we will define p63 dependent SG cellular identities and the defined cellular and molecular signature that is associated with regeneration and in response to irradiation by performing single cell RNA-sequencing (Aim2). These studies will better elucidate the role of ΔNp63 in SG organogenesis, and adult gland maintenance, and elucidate its contribution towards SG regeneration and in response to irradiation induced injury. Importantly, our genetic and genomic studies will also uncover novel ΔNp63-pathways dependent and independent biomarkers and drivers of the distinct cell states associated with regeneration and radiosensitivity. Long term, knowledge garnered from our proposed mechanistic studies will have clinical and therapeutic implications for human patients who suffer from SG dysfunction diseases.

项目摘要 自我更新，增殖和分化之间的复杂和动态平衡 必须严格调节唾液腺（SG）的茎/祖细胞，以确保正确形态发生， 稳态和再生。正常SG功能的改变，在临床上经常表现为 脱卵泡与外胚层发育不良，Sjögren综合征以及γ-等疾病有关 口腔癌患者的辐照疗法。当前，造成损失的治疗方案仍然有限。 因此，确定控制茎/祖细胞函数的关键转录和信号网络 SG非常需要SG来促进靶向干细胞和基于再生的疗法。 ΔNP63，一个干细胞 富集的转录因子，在上皮再生功能中起关键作用，作为来自 ΔNP63-NULL动物的表型，暴露了发育停滞和上皮器官的产生 包括SG。但是，我们目前对ΔNP63基因的分子机制的了解 STEM/祖先的承诺，维护和差异所需的表达程序 缺乏SG中的细胞人群。这是确定p63驱动的监管网络，特别是在 全球和基因组环境是迈向更好地理解SG STEM/祖细胞生物学的关键步骤 细胞并最终指导新的策略治疗SG功能障碍。为了解决这些知识差距， 我们将利用多种通用的小鼠模型来研究两个主要的独立领域。首先，我们会的 使用条件敲除小鼠模型检查ΔNP63在SG形态发生和成年中的作用 组织维护和修复（AIM1）。这种系统的研究急需，因为它们将确定 第一次，p63在SG发育和编排茎/祖细胞分化中的功能作用 程序。其次，我们将使用p63敲除鼠标模型和谱系跟踪实验来确定 p63+茎和祖细胞在SG再生过程中的贡献，并响应辐射诱导 损害。此外，我们将定义依赖p63的SG细胞身份以及定义的细胞和 与再生和响应辐照相关的分子特征通过执行单细胞 RNA-severing（AIM2）。这些研究将更好地阐明ΔNP63在SG器官发生和成人中的作用 腺维护，并阐明其对SG再生的贡献和对辐照的响应 诱发受伤。重要的是，我们的遗传和基因组研究还将发现新颖的ΔNP63-Pathway 与再生相关的不同细胞状态的依赖和独立的生物标志物和驱动因素 放射敏性。从长远来看，我们提出的机械研究获得的知识将具有临床和 对患有SG功能障碍疾病的人类患者的治疗意义。