Genetics and Pathobiology of Disorders of Keratinization

项目总结： 角化异常(DOK)是一种严重的、罕见的遗传性皮肤病，在这种皮肤病中， 皮肤的屏障功能被破坏，过度增殖的代偿途径是 在局部区域或整个皮肤表面激活。这导致了严重的 受损的表皮未能充分保护皮肤免受细菌、化学物质和 机械冲击或防止经皮水分流失。在早期生活中， 这些疾病可能危及生命，因为水泡会增加感染的易感性。 和裂缝，由于蒸发热损失和 表皮周转率增加，以及各种相关的并存情况和全身 可能会持续一生的异常现象。我们和其他人已经确定了70多种不同的 这些疾病的基因，然而临床经验和我们的数据表明，这些基因可以解释 只有一部分DOK的遗传力，表现出明显的基因座和表型 异质性，超过15%的受试者没有已知基因突变。在最后一次 在资助期内，我们发现了5个新的DOK基因，为表型提供了证据 在另外两个疾病中进行了扩展，并确定了一种针对一种疾病的新的发病机制导向疗法。 我们现在建议扩大我们大量表型良好的朝鲜血统的队列，以筛选 已知致病基因的突变，并使用外显子组和基因组测序 未发现已知DOK基因突变的受试者发现这些疾病的新遗传原因 精神错乱。我们将使用患者来源的细胞和组织来询问 发现了新的基因，并将产生转基因皮肤等价物来研究和证明 已识别突变的发病机制。有限数量的新基因之前没有 与DOK有关，我们将使用CRISPR技术生成鼠标模型，以进一步 检查已识别的突变对皮肤功能的影响。这些研究将继续 确定复杂的表皮分化和分化过程的核心分子途径 自我更新，并将为未来更详细的生物学研究提供关键背景。

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.