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.

项目摘要 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 辐射和黑素细胞干细胞在生命中维持健康的黑素细胞。失调 黑素细胞（MCS）或黑素细胞干细胞（MCSC）导致疾病显着影响 个人的生活质量，包括过度和低调的疾病以及致命的癌症。现在， 关于许多色素沉着障碍的分子机制或控制 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 MCSC/MC，并探讨如何为预防和治疗目的操纵MCSC/MC。 这里的目的是研究MC谱系的增殖，分化和维护 解剖SASH1及其相关蛋白的功能。我们建议的理由是Sash1 是一种小说，且不充分特征在于MC/MCSC和色素沉着，因此 了解其相关功能可能会导致新的潜在方法和操纵目标 MCSC/MC治疗与MC相关的疾病。 我们确定Sash1（S519N）中的一个突变是遗传性过度色素沉着障碍的原因。 根据我们的数据和文献，我们假设SASH1在增殖中起着至关重要的作用， 黑素细胞谱系的分化和维持。我们还建议SASH1S519N干扰 这些细胞的有丝分裂功能和分化，导致色素沉着障碍。测试我们的 假设，AIM 1将定义SASH1在增殖和分化中的分子机制 具有患者特异性诱导多能干细胞（IPSC）模型的MCSC/MC。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 带有CRISPR/CAS系统的克隆。使用IPSC，以及从中得出的MCSC和MCS 克隆，我们将定义SASH1变体的增殖和分化动力学，并检查 SASH1在黑素细胞谱系中功能的基础机制的基础。我们已经确定了 Sash1的几个约束伙伴。我们将进一步研究它们与Sash1角色的功能相关性 在具有遗传，生化，分子和细胞生物学方法的黑素细胞谱系中。目标2 将使用小鼠模型研究SASH1在维持黑素细胞谱系中的功能。我们将 测试SASH1损失如何使用SASH1基因敲除小鼠影响这些细胞的增殖和分化。 因此，我们希望我们的工作将对机制和新的新见解产生积极影响 MC和MCSC生物学领域的调节剂以及与这些细胞相关的疾病研究。