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(申请人提供)：角质形成细胞干细胞在维持表皮和毛囊的正常结构和功能方面具有毋庸置疑的作用，是遗传性和获得性皮肤病的重要参与者。因此，寻找调控其数量和增殖能力的基因是皮肤生物学中的一个关键问题。我们采用了一种新的基因鉴定策略，利用了我们实验室最近取得的几项进展，包括1)一种特别灵敏和定量的角质形成干细胞体外检测方法，2)发现这些克隆的数量和大小是数量上的多基因特征，以及3)对形成最大克隆的大量干细胞进行遗传定位。本研究的目的是在小鼠4号染色体上命名为角质形成细胞干细胞2(Ksc2)的一个基因或基因簇中确定一个基因或基因簇。我们的假设是，该基因中的一个或多个基因调节形成最大角质形成细胞集落的干细胞的数量。具体目标1的重点是使用更多的遗传和统计工具，将我们先前确定的Ksc2基因座的连锁间隔从大约10 cM缩小到大约0.5 cM。在具体目标2中，我们将使用候选基因方法，使用数据库挖掘和基于实验室的方法来识别干细胞基因。在目标3中，我们将使用互补基因表达谱来识别差异过表达或低表达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控多态。利用这三个目标的组合，交互作用的方法，我们期望识别和表征形成最大克隆的角质形成细胞干细胞的新的调控基因，因此，最高的增殖潜力。这个项目的意义在于，Ksc2基因座上的基因或基因簇，因为它与大量最大的菌落连锁，可能成为皮肤癌治疗和预防的一个令人惊讶和意想不到的目标。我们提出的方法代表了强大的遗传学方法的新应用，以一种全新的方式应用于一种全新的表型：角质形成细胞集落的数量和大小。我们以前发现克隆数反映了角质形成细胞干细胞增殖潜能的数量。这项拟议的研究将影响我们对上皮干细胞调控的理解。该项目不仅对了解毛囊和表皮的动态平衡具有基础科学意义，而且可能对皮肤癌、增生性皮肤病的诊断、预防和治疗以及干细胞功能下降的潜在情况具有临床影响。