Elucidating the role of p63 and transcriptional control mechanisms in progenitor cells of the salivary gland

阐明 p63 和转录控制机制在唾液腺祖细胞中的作用

• 批准号: 9243483
• 负责人: Rose-Anne Romano
• 金额: $ 11.91万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243483
• 关键词: Address; Adult; Animals; Area; Automobile Driving; Biological Assay; Biological Markers; Biological Models; Biology; Cell Culture Techniques; Cell Lineage; Cell physiology; Cells; ChIP-seq; Characteristics; Clinical; Critical Pathways; DNA; Development; Disease; Embryonic Development; Ensure; Epithelial; Epithelial Cells; Equilibrium; Foundations; Functional disorder; Future; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Targeting; Genes; Genetic; Genetic Transcription; Genomic approach; Genomics; Gland; Goals; Head and neck structure; Homeostasis; Human; Injury; Knockout Mice; Knowledge; Label; Light; Mammary gland; Maps; Molecular; Molecular Profiling; Morphogenesis; Mus; Natural regeneration; Normal tissue morphology; Organ; Organogenesis; Pathway interactions; Patients; Phenotype; Play; Population; Process; Property; Protein Isoforms; Regulator Genes; Role; Salivary Glands; Signal Pathway; Signal Transduction; Sjogren' s Syndrome; Skin; Squamous cell carcinoma; Stem cells; Submandibular gland; Systems Biology; Techniques; Technology; Therapeutic; Time; Tissue Engineering; Tissues; Transcriptional Regulation; Transgenic Mice; Xerostomia; base; cell type; epigenomics; experimental study; genetic approach; genome-wide; gland development; in vivo; injured; injury and repair; insight; interest; irradiation; mRNA Expression; mouse model; next generation sequencing; novel; novel therapeutic intervention; progenitor; prospective; regenerative; stem; stem cell biology; stem cell division; tissue regeneration; tool; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY The delicate balance between 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 Sjögren’s Syndrome and with γ-irradiation therapy of Head and Neck Squamous Carcinoma patients. Currently, treatment options for hyposalivation and mechanisms for restoring normal SG function remain limited. Therefore, the development of new tools and strategies directed at identifying transcriptional and signaling networks underlying stem/progenitor cell function of the SG with the ultimate goal for use in stem cell regenerative-based therapies and tissue engineering approaches, are critical. Hence, our goal is to examine the transcriptome landscape of the stem/progenitor cells of the SG and identify and characterize the regulatory networks of this cell population. It is well established that ΔNp63, plays a critical role in epithelial regenerative function as ΔNp63-null animals fail to develop several epithelial-rich organs including the SG. However, our current knowledge of how ΔNp63 regulates gene expression in the stem/progenitor cells of the SG is lacking. Thus examining the p63-driven regulatory networks is a key step towards a better understanding of the biology of the SG progenitor cells and in directing new strategies in treating SG dysfunction. To address these knowledge gaps, we will utilize SG stem/progenitor cells obtained from novel ΔNp63-GFP transgenic mice to study two major areas of interest. First, we will perform functional assays to compare the abilities of ΔNp63- GFPhi and ΔNp63-GFPlow SG cells to retain both their progenitor capacity and their ability to differentiate into mature cell lineages in SG organospheres (Aim1A). Furthermore, we will perform in vivo transcriptomic profiling (RNA-seq) to generate the mRNA expression profile in a pure population of ΔNp63-GFPhi SG progenitor cells and ΔNp63-GFPlow SG cells (Aim 1B). Such in vivo studies are important, since they will identify for the first time the gene expression profile of SG progenitor cells on a broad and dynamic scale. Second, we will perform in vivo ChIP-seq studies to identify key regulatory networks and pathways regulated by ΔNp63 in SG cells. Collectively, our approach using a genetically-defined model system and cutting-edge next generation sequencing technology will better elucidate the transcriptomic landscape of SG progenitor cells and shed light on the ΔNp63-governed transcriptional regulatory network and signaling pathways. Since such genomic studies are particularly lacking for SG cells, our proposed experiments will likely reveal new biomarkers and important regulators and drivers of stem cell function in this organ. Long term, such knowledge will have clinical and therapeutic implications for human patients who suffer from SG dysfunction diseases.

项目摘要 唾液的茎/祖细胞的增殖与分化之间的微妙平衡 必须严格调节腺体（SG），以确保适当的形态发生，稳态和再生。 正常SG功能的改变，通常在临床上表现出疾病，与疾病有关 例如Sjögren综合征以及对头部和颈部鳞状癌患者的γ辐射疗法。 当前，恢复正常SG功能的降低和机制的治疗方案仍然存在 有限的。因此，开发旨在识别转录和的策略的新工具和策略 SG的STEM/祖细胞功能的信号网络具有最终目标 基于再生的疗法和组织工程方法至关重要。因此，我们的目标是检查 SG的茎/祖细胞的转录组景观，并识别和表征调节 该细胞种群的网络。可以很好地确定ΔNP63在上皮再生中起着至关重要的作用 作为ΔNP63-NULL动物的功能无法开发包括SG在内的几种富含上皮的器官。但是，我们的 目前缺乏ΔNP63如何调节SG茎/祖细胞中基因表达的知识。 检查p63驱动的监管网络是更好地理解生物学的关键一步 SG祖细胞和指导治疗SG功能障碍的新策略。解决这些 知识差距，我们将利用从新型ΔNP63-GFP转基因小鼠获得的SG茎/祖细胞到 研究两个主要感兴趣的领域。首先，我们将执行功能性攻击以比较Δnp63-的能力 GFPHI和ΔNP63-GFPLOW SG细胞既保留其祖细胞的容量，又可以分化为 SG组织层中成熟的细胞谱系（AIM1A）。此外，我们将执行体内转录组 分析（RNA-SEQ）在纯ΔNP63-GFPHI SG中生成mRNA表达谱 祖细胞和ΔNP63-GFPLOW SG细胞（AIM 1B）。这样的体内研究很重要，因为它们会 首次识别SG祖细胞细胞的基因表达谱在广泛而动态的尺度上。 其次，我们将进行体内芯片序列研究，以确定受调节的关键调节网络和路径 由SG细胞中的ΔNP63。总的来说，我们使用一般定义的模型系统和尖端的方法 下一代测序技术将更好地阐明SG祖细胞的转录组景观 并在ΔNP63的转录调控网络和信号通路上亮起。自从这样 SG细胞尤其缺乏基因组研究，我们提出的实验可能会揭示新的 该器官中干细胞功能的生物标志物以及重要的调节剂和驱动因素。长期，这样的知识 对患有SG功能障碍疾病的人类患者具有临床和治疗意义。