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，可以保护我们的皮肤免受太阳紫外线辐射（UVR）的影响。我们发现了一种新基因 SASH1，它与遗传性色素沉着过度表型有关，该表型会增加黑色素细胞数量和皮肤色素沉着，并改变角质形成细胞的紫外线反应。 SASH1是一种潜在的肿瘤抑制因子；然而，对于 SASH1 在黑色素细胞、角质形成细胞或色素沉着中的功能知之甚少。因此，了解这种新颖且未表征的人类色素沉着基因的功能以及这种独特疾病的病因学将带来对黑素细胞、色素沉着和角质形成细胞生物学的新见解。根据我们对人类患者、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色素沉着以及黑素细胞的发育、分化和维持中发挥作用。为了检验我们的假设，我们将使用强大且互补的细胞培养系统和斑马鱼模型的创新组合。单细胞培养系统最适合研究单个细胞类型的生物学功能，而 3D 皮肤重建细胞培养系统最适合研究细胞类型之间的通讯，例如皮肤中的通讯。由于斑马鱼与哺乳动物具有保守的色素沉着途径、检测的可用性以及所有发育阶段的透明度，因此成为研究黑素细胞发育的最佳模型之一。对于这个R03，我们将重点研究野生型SASH1的功能，并为未来R01项目的详细研究建立协议、模型和初步数据。目标 1 将定义 SASH1 在黑素细胞和角质形成细胞中的作用。黑色素细胞和角质形成细胞对于皮肤色素沉着都很重要，我们的数据表明 SASH1 在这两种细胞类型中都起作用。我们将进一步：1）明确黑色素细胞中SASH1敲低对色素沉着、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哪些黑色素生成因子受 UVR 影响 SASH1 的调节； 3) 测试 SASH1 在体外模拟人体皮肤的 3D 皮肤重建系统中的作用。目标 2 将描述 sash1a 在斑马鱼黑色素细胞发育和再生中的功能。我们的临床数据表明，SASH1 调节黑素细胞再生，斑马鱼中 sash1a 的敲低会破坏黑素细胞胚胎发育。我们将进一步 1) 通过检查神经嵴祖细胞的规格、迁移和存活来测试 sash1a 敲低如何影响黑素细胞的发育； 2) 确定 sash1a 敲低是否影响黑素细胞特异性消融后的黑素细胞再生； 3) 使用 RNA-Seq 定义 sash1a 调控的途径。总之，我们将结合两种互补的模型来研究一种新颖且未表征的色素沉着基因的体外和体内功能。未来的 R01 应用将建立在 R03 初步数据的基础上，进一步阐明 SASH1 在皮肤紫外线防护方面的功能。