Popliteal Pterygium syndrome, IRf6, and the periderm

腘胬肉综合征、IRf6 和周皮

• 批准号: 10727050
• 负责人: MARTINE DUNNWALD
• 金额: $ 15.55万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10727050
• 关键词: Accounting; Adhesions; Affect; Alleles; Basal Cell; Biological Markers; Biology; Cell-Cell Adhesion; Cells; Characteristics; Choristoma; Cleft lip with or without cleft palate; Clinical; Conjunctival Pterygium; DNA Binding Domain; Defect; Development; Disease; Ectoderm; Embryo; Embryonic Development; Epidermis; Epithelial Cells; Etiology; Event; Exhibits; Eyelid structure; Fingers; Functional disorder; Future; Gene Expression Profile; Genetic; Genital; Genitalia; Genomic approach; Heterogeneity; Immunofluorescence Immunologic; In Situ; Interferons; Internet; Knee; Knowledge; Libraries; Live Birth; Molecular Profiling; Morphogenesis; Mus; Mutation; Online Mendelian Inheritance In Man; Oral cavity; Outcome; Periderm; Process; Psoriasis; Rare Diseases; Resolution; Role; Skin; Specific qualifier value; Squamous Epithelium; Stratification; Stratum Basale; Stratum corneum; Syndactyly; Testing; Time; Tissues; Validation; Work; adhesion receptor; design; gene regulatory network; innovation; insight; keratinization; keratinocyte; keratinocyte differentiation; novel; oral cavity epithelium; popliteal pterygium syndrome; prevent; single-cell RNA sequencing; skin organogenesis; transcription factor

SUMMARY This project focuses on popliteal pterygium syndrome (PPS, OMIM 119500), a rare disease occurring in 1/300,000 live births. It is estimated that 97% of PPS cases can be accounted for by mutations that affect residues of the DNA binding domain of the transcription factor interferon regulatory factor 6 (IRF6). Clinical manifestations of PPS include cleft lip with or without cleft palate, and a variety of tissue adhesions, including pterygium (tissue fusion or webbing) behind the knees (popliteal region), syndactyly, ankyloblepharon (fusion of eyelids), and fusions around the genital region. Absence of the periderm, a single layer of epithelial cells that covers the developing epidermis during embryogenesis, is believed to underlie these aberrant tissue adhesions. Relatively little is known about the periderm, and studies aimed at underlying the pathophysiology of PPS may provide new insight with broad significance for our understanding of how embryonic ectoderm gives rise to the rich complexity of skin and oral epithelia. We found that transcription factors implicated in epidermal differentiation, including IRF6, are expressed in the periderm. Yet, the field lacks in the availabiltity of biomarkers that specificially reflect the genesis, function and fate of periderm cells, accounting for our superficial understanding of how defects in the periderm may contribute to PPS. Established markers of the periderm are absent in the developing ectoderm of Irf6-deficient mice. In the latter setting, abnormal tissue adhesions occur in the oral cavity and differentiation of keratinocytes in the developing epidermis is compromised. Our prior studies also revealed that Irf6-deficient keratinocytes can stratify, but cannot give rise to periderm. While these observations establish that IRF6 is essential for specification of the periderm, how it affects morphogenesis of the periderm and how the periderm prevents tissue fusion remain ill-defined. The premise of our study is that a deeper understanding of PPS requires a much more profound understanding of the role of IRF6 in the formation of the periderm. Toward this end, we herein propose to define how the periderm initially forms, and identify the IRF6-dependent gene regulatory networks that contribute to this initial morphogenic event. To test our central hypothesis that periderm cells present a unique gene expression profile throughout its genesis and fate that are affected by IRF6 levels, we will generate cellular libraries from wild-type and Irf6-deficient epidermis obtained from embryos at different developmental time points critical for periderm morphogenesis and perform single-cell RNA sequencing, followed by multiplex in situ and immunofluorescence validation. This proposal is innovative both conceptually and technically as it represents a significant conceptual paradigm shift and takes a novel single-cell genomic approach to understanding cellular heterogeneity within the epidermis at single-cell resolution during early stages of morphogenesis. Our knowledge of, and expertise in IRF6 biology and skin development establish that we are well-prepared to succeed with all aspects of this challenge.

概括 该项目侧重于pterliteal pterygium综合征（PPS，Omim 119500），这是一种罕见疾病 1/300,000活产。据估计，有97％的PPS病例可以通过影响的突变来解释 转录因子干扰素调节因子6（IRF6）的DNA结合结构域的残基。临床 PPS的表现包括唇裂，​​有或不带left裂，以及各种组织粘连，包括 膝盖（Popliteal区域）后面的翼肉（组织融合或织带），syndactyly，ankyblyblepharon（融合 眼睑）和生殖区域周围的融合。缺乏Periderm，一层上皮细胞 涵盖胚胎发生过程中发育的表皮，据信是这些异常组织粘附的基础。 对periderm的了解相对鲜为人知，旨在为PPS的病理生理生物生理的研究可能 为我们了解胚胎外胚层如何产生的新见解具有广泛的意义 皮肤和口服上皮的丰富复杂性。我们发现与表皮有关的转录因子 包括IRF6在内的分化在Periderm中表达。然而，该领域缺乏生物标志物的可用性 该典型地反映了佩里德尔细胞的起源，功能和命运，占我们肤浅的 了解Periderm中的缺陷如何有助于PP。 Periderm已建立的标记是 在IRF6缺陷型小鼠的发育中不存在。在后一种情况下，发生异常的组织粘附 在发育表皮中角质形成细胞的口腔和分化中，表皮细胞受到损害。我们的先验 研究还表明，IRF6缺陷型角质形成细胞可以分层，但不能引起佩里德尔。而这些 观察结果表明，IRF6对于Periderm的规范至关重要，它如何影响形态发生 periderm以及如何防止组织融合保持不确定。我们研究的前提是 对PPS的更深入了解需要对IRF6在形成中的作用有更深刻的了解 Periderm。为此，我们在此提议定义Periderm最初的形成方式，并确定 IRF6依赖性基因调节网络有助于这一初始形态发生事件。测试我们的中央 佩里德尔细胞在整个起源和命运中呈现独特的基因表达谱的假说 受IRF6水平的影响，我们将产生来自野生型和IRF6缺陷表皮的细胞库 从不同发育时间点的胚胎对periderm形态发生至关重要并进行单细胞 RNA测序，然后进行多重原位和免疫荧光验证。该建议是创新的 从概念和技术上都在技术上都代表了一个重大的概念范式转变，并进行了小说 单细胞基因组方法了解单细胞在表皮内的细胞异质性 在形态发生的早期阶段的分辨率。我们对IRF6生物学和皮肤方面的知识和专业知识 发展确定，我们为在这项挑战的各个方面都做好了准备。