Genetics and Pathobiology of Disorders of Keratinization

项目概要： 角化障碍（DOK）是一种严重的、罕见的遗传性皮肤病， 皮肤的屏障功能被破坏， 在局部区域或整个皮肤表面被激活。这导致了严重的 受损的表皮不能充分保护免受细菌、化学物质和 机械冲击或防止经表皮水分流失。在早期生活中， 这些疾病可能危及生命，由于水泡而增加了感染的易感性， 和裂缝，由于蒸发热损失而急剧增加的代谢需求， 表皮更新率增加，以及各种相关的合并症和全身性疾病 这些异常可以持续一生。我们和其他人已经确定了70多种 这些疾病的基因，但临床经验和我们的数据表明，这些基因解释， DOK的遗传力只有一部分，表现出显著的位点和表型 异质性，超过15%的受试者在已知基因中没有突变。在过去 在资助期间，我们鉴定了五个DOK的新基因，为DOK的表型特征提供了证据。 在其他两个扩展，并确定了一种新的发病机制为导向的治疗疾病。 我们现在建议扩大我们的大队列良好的表型DOK激酶，筛选 已知致病基因的突变，并采用外显子组和基因组测序， 受试者在已知的DOK基因中没有突变，以发现这些疾病的新遗传原因。 紊乱我们将使用患者来源的细胞和组织来询问 确定了新的基因，并将产生转基因皮肤等效物，以研究和证明 确定突变的发病机制。对于有限数量的新基因， 与DOK有关，我们将使用CRISPR技术生成小鼠模型， 检查已鉴定的突变对皮肤功能的影响。这些研究将继续 识别表皮分化复杂过程的核心分子途径， 自我更新，并将提供更详细的未来生物学研究的关键背景。

PROJECT SUMMARY: Disorders of keratinization (DOK) are severe, rare genetic skin disorders in which the central barrier function of the skin is disrupted and a compensatory pathway of hyper-proliferation is activated in either a localized area or throughout the entire skin surface. This results in a grossly compromised epidermis that fails to adequately protect against bacterial, chemical, and mechanical assault or to prevent transepidermal water loss. In early life, the consequences of these disorders can be life-threatening, with increased susceptibility to infection due to blistering and fissures, dramatically increased metabolic demands due to evaporative heat loss and increased epidermal turnover rate, and various associated comorbid conditions and systemic abnormalities which can persist throughout life. We and others have identified over 70 diverse genes for these disorders, yet clinical experience and our data show that these genes explain only a portion of heritability for DOK, which demonstrate marked locus and phenotypic heterogeneity, with greater than 15% of subjects without mutation in known genes. In the last funding period, we identified five new genes for DOK, provided evidence for phenotypic expansion in two others, and identified a novel pathogenesis-directed therapy for one disorder. We now propose to expand our large cohort of well-phenotyped DOK kindreds, to screen for mutations in known causative genes, and to employ exome and genome sequencing in those subjects without mutation in known DOK genes to discover novel genetic causes of these disorders. We will employ patient-derived cells and tissue to interrogate the function of identified novel genes, and will generate transgenic skin equivalents to study and prove pathogenesis of identified mutations. For a limited number of novel genes not previously implicated in DOK, we will generate mouse models using CRISPR technology to further examine the effect of identified mutations on cutaneous function. These studies will continue to identify molecular pathways central to the complex processes of epidermal differentiation and self-renewal, and will provide critical context for more detailed future biologic studies.