Identification of a Keratinocyte Stem Cell Regulatory Gene in the KSC2 Locus

KSC2 基因座中角质形成细胞干细胞调节基因的鉴定

• 批准号: 8707971
• 负责人: REBECCA Jane MORRIS
• 金额: $ 33.73万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707971
• 关键词: Alleles; Basic Science; Biological Assay; Biology; Candidate Disease Gene; Cell Count; Cell physiology; Cells; Characteristics; Chromosome Mapping; Chromosomes, Human, Pair 4; Chromosomes, Human, Pair 9; Clinical; Code; Colony-Forming Units Assay; Cutaneous; Databases; Development; Diagnosis; Epidermis; Epithelial; Gene Chips; Gene Cluster; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Targeting; Genes; Genetic; Genetic Polymorphism; Hair follicle structure; Homeostasis; Hyperplasia; In Vitro; Inherited; Knockout Mice; Knowledge; Label; Laboratories; Large Keratinocyte; Light; Link; Malignant Neoplasms; Maps; Methods; Mining; Molecular; Mus; Names; Pathway interactions; Phenotype; Predisposition; Prevention; Quantitative Trait Loci; Regulation; Regulator Genes; Research; Role; Skin; Skin Cancer; Skin Neoplasms; Stem cells; Structure; Transgenic Mice; Variant; Work; base; cancer prevention; cancer therapy; computerized tools; in vitro Assay; in vivo; keratinocyte; mouse model; mutant; novel; novel strategies; response; skin disorder; stem; toll-like receptor 4; tool; trait

DESCRIPTION (provided by applicant): Keratinocyte stem cells have an unquestioned role in maintaining the normal structure and function of the epidermis and hair follicles and are important players in inherited and acquired skin diseases. Therefore, identification of genes regulating their numbers and proliferative potential is a critical problem in cutaneous biology. We have taken a novel strategy for gene identification taking advantage of several recent advances made by our laboratory including 1) a particularly sensitive and quantitative in vitro assay for keratinocyte stem cells, 2) discovering that the number and size of those colonies are quantitative multigenic traits, and 3) genetically mapping a locus linked to a high number of stem cells forming the largest colonies. The objective of this proposed research is to identify a gene or gene cluster in a locus we have named Keratinocyte stem cell locus 2 (Ksc2) on mouse chromosome 4. Our hypothesis is that a gene (or genes) in this locus regulates the number of stem cells forming the largest keratinocyte colonies. The focus of Specific Aim 1 is to use additional genetic and statistical tools to narrow the linkage interval of the Ksc2 locus that we have previously identified from approximately 10 cM to approximately 0.5 cM. In Specific Aim 2, we will use a candidate gene approach for stem cell gene identification using database mining and lab-based methods. In Aim 3, we will use complementary gene expression profiling to identify differentially over- or under-expressed genes as well as associated molecular pathways, and to assess regulatory polymorphisms causing differences in gene expression. Using the combined, interactive approaches of these three aims, we expect to identify and characterize a new regulatory gene for the keratinocyte stem cells forming the largest colonies, and hence, the highest proliferative potential. The significance of this project is that the gene or gene cluster in the Ksc2 locus, because it shows linkage with a high number of the largest colonies, may be a surprising and unanticipated target for skin cancer therapy and prevention. The approach we present represents a novel application of powerful genetics methods applied in a completely new way to a completely new phenotype: the number and size of keratinocyte colonies. We previously found that colony number reflected the number of proliferative potential of keratinocyte stem cells. This proposed research will impact our understanding of epithelial stem cell regulation. This project not only has basic science ramifications for understanding hair follicle and epidermal homeostasis, it may have clinical impact for the diagnosis, prevention, and treatment of skin cancer, hyperplastic skin disease, as well as potential conditions of declining stem cell function.

描述（由申请人提供）：角质形成细胞细胞在维持表皮和毛囊的正常结构和功能中毫无疑问的作用，并且是遗传和获得的皮肤疾病的重要参与者。因此，鉴定调节其数量的基因和增生潜力是皮肤生物学的关键问题。我们采取了一种新的基因鉴定策略，利用了我们实验室的最新进展，包括1）对角质形成细胞细胞的特别敏感和定量的体外测定法，2）发现这些菌落的数量和大小是定量的多基因性状，以及3）基因映射到一个含有较高数量的干细胞的基因映射的，这是较高的colon colonies。这项拟议的研究的目的是在小鼠4染色体上鉴定我们称为角质形成细胞干细胞基因座2（KSC2）的基因群中的基因或基因簇。我们的假设是，该基因座中的基因（或基因）调节形成最大角膜细胞菌落的干细胞数量。特定目标1的重点是使用其他遗传和统计工具来缩小我们以前从大约10 cm到约0.5 cm的KSC2基因座的连锁间隔。在特定目标2中，我们将使用数据库挖掘和基于实验室的方法使用候选基因方法来识别干细胞基因。在AIM 3中，我们将使用互补的基因表达分析来鉴定差异过表达或不足的基因以及相关的分子途径，并评估导致基因表达差异的调节性多态性。使用这三个目标的组合，互动方法，我们希望鉴定并表征形成最大菌落的角质形成细胞干细胞的新调节基因，因此，增殖潜力最高。该项目的意义在于，KSC2基因座中的基因或基因簇，因为它与大量最大菌落的联系可能是对皮肤癌治疗和预防的令人惊讶且意外的目标。我们提出的方法代表了以一种全新的方式应用于全新表型的新型遗传学方法的新应用：角质形成菌群的数量和大小。我们先前发现，菌落数反映了角质形成细胞细胞的增殖潜力数量。这项提出的研究将影响我们对上皮干细胞调节的理解。该项目不仅具有了解毛囊和表皮稳态的基础科学影响，还可能对皮肤癌的诊断，预防和治疗产生临床影响，增生性皮肤病以及干细胞功能下降的潜在条件。