Study of melanocyte lineage through SASH1 and associated proteins

通过 SASH1 和相关蛋白研究黑素细胞谱系

• 批准号: 10596208
• 负责人: Yiqun G Shellman
• 金额: $ 38.47万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-13 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596208
• 关键词: Affect; Aging; Binding; Biochemical; Biological; Biological Process; Biology; Cancer Biology; Cell Cycle Progression; Cell Differentiation process; Cell Lineage; Cell Maintenance; Cell physiology; Cells; Clone Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Data; Disease; Genes; Genetic; Goals; Hair; Hyperpigmentation; Hypopigmentation; Impairment; In Vitro; Individual; Inherited; Kinetics; Knockout Mice; Life; Literature; Maintenance; Malignant Neoplasms; Measurable; Mediating; Mission; Mitotic; Mitotic spindle; Molecular; Mus; Mutation; National Institute of Arthritis, and Musculoskeletal, and Skin Diseases; Natural regeneration; Outcome; Pathologic; Pathology; Patients; Pattern; Phenotype; Pigmentation Disorders; Pigmentation physiologic function; Play; Population; Preventive; Proliferating; Proteins; Quality of life; Regenerative Medicine; Regulation; Research; Role; Structure; Study models; Subcutaneous Tissue; TNKS gene; Tankyrase; Testing; Therapeutic; Ultraviolet Rays; Variant; Vitiligo; Work; gene correction; in vivo; induced pluripotent stem cell; induced pluripotent stem cell technology; insight; interest; melanocyte; melanoma; mouse model; new therapeutic target; novel; novel strategies; premature; stem cell biology; stem cell model; stem cells; three-dimensional modeling; tool; young adult

PROJECT SUMMARY Pigmenting cells (melanocytes) protect vital stem cells and subcutaneous tissues from ultraviolet radiation, and melanocyte stem cells sustain a healthy population of melanocytes through life. Dysregulation of melanocytes (MCs) or melanocyte stem cells (McSCs) leads to diseases that significantly affects an individual's quality of life, including hyper- and hypo-pigmentation disorders, and fatal cancer. Currently, much is still undefined about the molecular mechanisms for many pigmentation disorders or for controlling melanocytes or melanocyte stem cells. Our long-term goal is to better understand the biology and pathology of McSC/MC, and to explore how McSC/MC can be manipulated for preventive and therapeutic purpose. The objective here is to study the proliferation, differentiation and maintenance of the MC lineage through dissecting the functions of SASH1 and its associated proteins. The rationale for our proposal is that SASH1 is a novel and not well characterized gene involved in MC/McSC and pigmentation, and therefore understanding its relevant functions will likely lead to new potential approaches and targets for manipulating McSC/MC to treat MC-related diseases. We identified a mutation in SASH1 (S519N) as causative for an inherited hyper-pigmentation disorder. Based on our data and literature, we hypothesize that SASH1 plays a critical role in proliferation, differentiation and maintenance of the melanocyte lineage. We also propose that SASH1S519N disrupts mitotic function and differentiation of these cells, resulting in the pigmentation disorder. To test our hypotheses, Aim 1 will define the molecular mechanisms of SASH1 in proliferation and differentiation of McSC/MC with the patient-specific induced pluripotent stem cell (iPSC) model. We will generate patient- specific iPSC clones from SASH1S519N individuals, and then construct gene-corrected and null isogenic clones with the CRISPR/Cas system. Using the iPSCs, as well as McSCs and MCs derived from these clones, we will define the proliferation and differentiation kinetics of SASH1 variants, and exam the underpinning molecular mechanism for SASH1's functions in melanocyte lineage. We have identified several binding partners for SASH1. We will further investigate their functional relevance to SASH1's roles in the melanocyte lineage with genetic, biochemical, molecular, and cellular biological approaches. Aim 2 will use mouse model to study the function of SASH1 in maintenance of the melanocyte lineage. We will test how SASH1 loss affects proliferation and differentiation of these cells using SASH1 knockout mice. Thus, we expect our work will have a positive impact with new insights into the mechanisms and new regulators in the fields of MC and McSC biology along with diseases research associated with these cells.

项目概要 色素细胞（黑素细胞）保护重要的干细胞和皮下组织免受紫外线的侵害 辐射和黑素细胞干细胞在一生中维持健康的黑素细胞群。失调 黑素细胞 (MC) 或黑素细胞干细胞 (McSC) 的减少会导致严重影响身体健康的疾病 个人的生活质量，包括色素沉着过度和色素减退性疾病以及致命的癌症。现在， 关于许多色素沉着疾病或控制的分子机制仍不清楚 黑素细胞或黑素细胞干细胞。我们的长期目标是更好地了解生物学和病理学 McSC/MC 的研究，并探索如何操纵 McSC/MC 来达到预防和治疗目的。 这里的目的是通过以下方式研究 MC 谱系的增殖、分化和维持： 剖析 SASH1 及其相关蛋白的功能。我们提议的理由是 SASH1 是一种涉及 MC/MCSC 和色素沉着的新型且尚未充分表征的基因，因此 了解其相关功能可能会带来新的潜在操纵方法和目标 McSC/MC 用于治疗 MC 相关疾病。 我们发现 SASH1 (S519N) 的突变是遗传性色素沉着过度症的病因。 根据我们的数据和文献，我们假设 SASH1 在增殖中发挥着关键作用， 黑素细胞谱系的分化和维持。我们还建议 SASH1S519N 中断 这些细胞的有丝分裂功能和分化，导致色素沉着障碍。来测试我们的 假设，目标 1 将定义 SASH1 在细胞增殖和分化中的分子机制 McSC/MC 与患者特异性诱导多能干细胞 (iPSC) 模型。我们将产生患者- 来自 SASH1S519N 个体的特定 iPSC 克隆，然后构建基因校正和无效同基因 使用 CRISPR/Cas 系统进行克隆。使用 iPSC 以及源自这些的 McSC 和 MC 克隆，我们将定义 SASH1 变体的增殖和分化动力学，并检查 SASH1 在黑素细胞谱系中的功能的基础分子机制。我们已经确定 SASH1 的几个结合伙伴。我们将进一步研究它们与 SASH1 角色的功能相关性 通过遗传、生化、分子和细胞生物学方法研究黑素细胞谱系。目标2 将使用小鼠模型来研究SASH1在维持黑素细胞谱系中的功能。我们将 使用 SASH1 敲除小鼠测试 SASH1 丢失如何影响这些细胞的增殖和分化。 因此，我们期望我们的工作能够通过对机制的新见解和新的内容产生积极的影响。 MC 和 McSC 生物学领域的调节器以及与这些细胞相关的疾病研究。