Aldehyde dehydrogenase: A novel regulator of melanin biogenesis

醛脱氢酶：黑色素生物发生的新型调节剂

基本信息

批准号：
8424830
负责人：
Anand K Ganesan
金额：
$ 12.39万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-01 至 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8424830
关键词：
4 hydroxynonenal Aldehydes Anabolism Apoptosis Biochemical Biogenesis Biological Assay Biology Cells Chemicals Coupled Couples Data Defect Elements Environment Enzymes Epidermis Gene Targeting Genes Genetic Genetic Transcription Glutathione Glutathione Disulfide Goals High Pressure Liquid Chromatography Human Individual Instruction Investigation Lipid Peroxidation Lipids Maps Measures Mediating Mediator of activation protein Melanins Melanogenesis Melanoma Cell Membrane Modeling Molecular Monophenol Monooxygenase Pigmentation physiologic function Pigments Principal Investigator Production RNA Interference Reaction Reactive Oxygen Species Regulation Resistance Skin Skin tanning Small Interfering RNA Technology Time UV carcinogenesis UV induced UVA induced UVB induced Ultraviolet Rays aldehyde dehydrogenase 1 aldehyde dehydrogenases base chromatin immunoprecipitation eumelanin genome-wide imaging modality indium arsenide inhibitor/antagonist interest keratinocyte lipid metabolism loss of function mRNA Expression melanin inhibitor melanocyte microphthalmia-associated transcription factor model development novel oxidized lipid pheomelanin prevent promoter research study response screening small hairpin RNA tissue culture transcription factor ultraviolet damage ultraviolet irradiation validation studies

项目摘要

DESCRIPTION (provided by applicant): While genetic and biochemical investigations have led to the identification of genes that regulate pigment production, the mechanisms by which melanocytes couple UV irradiation to melanin production has not been identified. Genome wide siRNA screening was performed to identify human genes that regulate melanogenesis in melanocytes and melanoma cells. Validation studies determined that this novel, unbiased approach had a low false positive and off-target rate, and identified novel genes that regulate melanin secretion and melanin biosynthesis. Two validated gene targets identified in this analysis, aldehyde dehydrogenase 1 and aldehyde dehydrogenase 9, are cellular genes involved in metabolizing byproducts of lipid peroxidation reactions. Preliminary data indicates that Aldhl and AldhQ impact the expression of tyrosinase and MITF in normal melanocytes and melanoma cells. The overall goal of this proposal is to understand the mechanism by which Aldhl and Aldh9 regulate melanin biosynthesis. Specific aim 1 will determine the mechanism by which Aldhl and AldhQ regulate tyrosinase and MITF protein levels. Specific aim 2 couples siRNA based loss of function approaches with chemical approaches to quantitate melanin to determine if Aldh activity impacts pheomelanin production. Specific aim 3 will couple loss of function approaches with standard plate assays to determine whether Aldh activity is required for UVA and UVB induced pigment production. Followup experiments will determine if Aldhl and Aldh9 maintain a permissive environment for melanin production by maintaining low intracellular levels of reactive oxygen species and lipid peroxidation products. Specific aim 4 will use novel imaging modalities and shRNA loss of funciton technology to determine whether Aldhl and Aldh9 impact pigment production in a human skin equivalent model. Through these studies, we will determine the mechanism by which Aldhl and Aldh9 regulate melanin production and gain a more complete understanding of the molecular mediators of UV induced pigment production. RELEVANCE (See instructions): UV induced melanin production protects the skin from the carcinogenic effects of UV light. While core genes that regulate pigment production have been identified, little is understood about how melanocytes produce more melanin in response to UV. In this proposal, we characterize putative regulators (Aldhl and Aldh9) of UV induced tanning and determine whether these genes regulate tanning in human skin models.

描述（由申请人提供）：虽然遗传和生化研究导致了调节色素产生的基因的鉴定，但尚未鉴定出黑色素对紫外线辐照到黑色素产生的机制。进行了基因组宽的siRNA筛选，以鉴定调节黑色素细胞和黑色素瘤细胞中黑色素生成的人类基因。验证研究确定，这种新颖的，公正的方法的假阳性和脱靶速率低，并确定了调节黑色素分泌和黑色素生物合成的新型基因。在该分析中鉴定出的两个经过验证的基因靶标，醛脱氢酶1和醛脱氢酶9是参与脂质过氧化反应代谢副产物的细胞基因。初步数据表明ALDHL和ALDHQ在正常的黑色素细胞和黑色素瘤细胞中影响酪氨酸酶和MITF的表达。该提案的总体目标是了解ALDHL和ALDH9调节黑色素生物合成的机制。具体目标1将确定ALDHL和ALDHQ调节酪氨酸酶和MITF蛋白水平的机制。具体目标2夫妇用化学方法基于siRNA的功能方法来定量黑色素，以确定ALDH活性是否会影响苯烷素的产生。特定的目标3将损失功能方法与标准板测定法，以确定UVA和UVB诱导色素产生是否需要ALDH活性。随访实验将确定ALDHL和ALDH9是否通过维持低细胞内的活性氧和脂质过氧化产物来维持黑色素产生的允许环境。特定目标4将使用新颖的成像方式和shRNA损失Funciton技术来确定ALDHL和ALDH9是否影响人类皮肤等效模型中的色素产生。通过这些研究，我们将确定ALDHL和ALDH9调节黑色素的产生的机制，并对紫外线诱导的色素产生的分子介体获得更完整的了解。相关性（请参阅说明）：紫外线诱导的黑色素产生可保护皮肤免受紫外光的致癌作用。尽管已经鉴定出调节色素生产的核心基因，但关于黑素细胞如何产生更多黑色素的反应，几乎没有理解。在此提案中，我们表征了紫外线诱导的晒黑的推定调节剂（Aldhl和Aldh9），并确定这些基因是否调节了人类皮肤模型中的晒黑。