The role of SASH1 in skin pigmentation

SASH1在皮肤色素沉着中的作用

基本信息

批准号：
9056438
负责人：
Yiqun G Shellman
金额：
$ 7.78万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9056438
关键词：
Ablation Affect Aging Area Biological Assay Biological Models Biological Process Biology Cell Aging Cell Count Cell Culture System Cell Culture Techniques Cell Differentiation process Cell Proliferation Cell Survival Cells Clinical Data Communication Data Development Disease Embryonic Development Environment Etiology Family Future Genes Goals Health Human Hyperpigmentation Hypopigmentation Immigration In Vitro Individual Inherited Investigation Knowledge Lead Lentigo Lesion Maintenance Malignant Neoplasms Mammals Modeling Mono-S Mutate Mutation Natural regeneration Nature Neural Crest Pathway interactions Patients Phenotype Physical Carcinogens Pigmentation Disorders Pigmentation physiologic function Pigments Play Preparation Prevention Prevention strategy Proteins Protocols documentation RNA Radiation Research Role Signal Transduction Skin Skin Cancer Skin Pigmentation Solar Lentigo Spottings Staging Stress Study models Testing Tumor Suppressor Proteins UV induced UV response Ultraviolet Rays Viola Zebrafish adapter protein base cancer cell cell growth cell type in vivo innovation insight keratinocyte knock-down melanocyte member migration mutant novel premature progenitor response skin disorder three-dimensional modeling

项目摘要

DESCRIPTION (provided by applicant): Melanocytes are pigment producing cells that protect our skin from solar ultraviolet radiation (UVR). We identified a novel gene, SASH1, involved in an inherited hyperpigmentation phenotype, with increased melanocyte number and pigmentation in skin and altered UV response of keratinocytes. SASH1 is a potential tumor suppressor; however, not much is known about the functions of SASH1 in melanocytes, keratinocytes, or pigmentation. Thus, understanding the function of this novel and uncharacterized human pigmentation gene and etiology of this unique disorder will lead to new insight in melanocyte, pigmentation, and keratinocyte biology. Based on our preliminary results from human patients, in vitro and zebrafish, we hypothesize that SASH1 functions in UV-induced pigmentation involving melanocytes and keratinocytes, as well as in the development, differentiation and maintenance of melanocytes. To test our hypotheses, we will use an innovative combination of powerful and complementary cell culture system and zebrafish models. Mono-cell culture systems are best to study the biological functions in individual cell types, and 3D skin- reconstruct cell culture systems are best to study communication between cell types, such as in skin. Due to their conserved pigmentation pathways with mammals, the availability of assays, and the transparency of all development stages, zebrafish is one of the best models for studying development of melanocytes. For this R03, we will focus on studying the functions of wild type SASH1 and establishing protocols, models, and preliminary data for future detailed investigations in a R01 project. Aim 1 will define the roles of SASH1 in melanocytes and keratinocytes. Both melanocytes and keratinocytes are important for skin pigmentation, and our data suggest that SASH1 functions in both cell types. We will further: 1) define the effects of SASH1 knockdown in melanocytes on pigmentation, differentiation, cellular aging, cell growth and survival; 2) determine which melanogenic factor(s) are regulated by SASH1 upon UVR in keratinocytes; and 3) test SASH1's role in a 3D skin reconstruct system for modeling human skin in vitro. Aim 2 will characterize the function of sash1a in the development and regeneration of melanocytes in zebrafish. Our clinical data suggest that SASH1 regulates melanocyte regeneration, and sash1a knock-down in zebrafish disrupts melanocyte embryonic development. We will further 1) test how sash1a knockdown affects melanocyte development, by examining neural crest progenitor specification, migration, and survival; 2) determine whether sash1a knockdown affects melanocyte regeneration following melanocyte-specific ablation; and 3) define the pathways regulated by sash1a using RNA-Seq. In summary, we will combine two complementary models to study the function of a novel and uncharacterized pigmentation gene in vitro and in vivo. A future R01 application will build on R03 preliminary data to further elucidate the function of SASH1 in the UV protective nature of skin.

描述（由申请人提供）：黑素细胞是产生色素的细胞，可保护我们的皮肤免受太阳紫外线辐射（UVR）的影响。我们确定了一种新型基因SASH1，涉及遗传性色素沉着表型，皮肤中的黑素细胞数和色素沉着增加，并改变了角质形成细胞的紫外线反应。 Sash1是一种潜在的肿瘤抑制剂。然而，关于sash1在黑素细胞，角质形成细胞或色素沉着中的功能知之甚少。因此，了解这种独特疾病的这种新颖和未表征的人类色素沉着基因的功能将导致黑素细胞，色素沉着和角质形成细胞生物学的新见解。根据我们的体外和斑马鱼的初步结果，我们假设SASH1在涉及黑素细胞和角质形成细胞的紫外线诱导的色素沉着中以及在黑素细胞的发育，分化和维持中的功能。为了检验我们的假设，我们将使用功能强大和互补的细胞培养系统和斑马鱼模型的创新组合。单细胞培养系统最好研究单个细胞类型中的生物学功能，而3D皮肤 - 重建细胞培养系统最好研究细胞类型（例如皮肤）之间的通信。由于它们具有哺乳动物的保守色素沉着途径，分析的可用性以及所有发育阶段的透明度，斑马鱼是研究黑素细胞发育的最佳模型之一。对于此R03，我们将专注于研究野生型SASH1的功能，并在R01项目中为将来的详细研究建立协议，模型和初步数据。 AIM 1将定义SASH1在黑素细胞和角质形成细胞中的作用。黑素细胞和角质形成细胞对皮肤色素沉着都很重要，我们的数据表明SASH1在两种细胞类型中都起作用。我们将进一步：1）定义黑素细胞中Sash1敲低的影响对色素化，分化，细胞衰老，细胞生长和存活的影响； 2）确定角质形成细胞中UVR对SASH1调节哪种黑色素生成因子； 3）测试SASH1在3D皮肤重建系统中的作用，用于在体外对人皮肤进行建模。 AIM 2将表征SASH1A在斑马鱼中黑素细胞发展和再生中的功能。我们的临床数据表明，SASH1调节黑素细胞再生，而在斑马鱼中的SASH1A敲除了破坏黑素细胞的胚胎发育。我们将进一步1）测试SASH1A敲低如何通过检查神经Crest祖细胞规范，迁移和生存来影响黑素细胞的发育； 2）确定sash1a敲低是否影响黑色素特异性消融后黑素细胞再生； 3）使用RNA-Seq定义由SASH1A调节的途径。总而言之，我们将结合两个互补模型，以研究一种新型和未表征的色素沉着基因在体外和体内的功能。未来的R01应用程序将基于R03初步数据，以进一步阐明SASH1在皮肤的紫外线保护性中的功能。