Elucidation of Arvcf-dependent mechanisms required for lens function

阐明晶状体功能所需的 Arvcf 依赖性机制

基本信息

批准号：
10615869
负责人：
Timothy F Plageman
金额：
$ 39.68万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10615869
关键词：
ARVCF gene Address Adherens Junction Adhesions Age Months Animal Model Architecture Attenuated Bilateral Binding Biochemical Biological Models Biology Biomechanics Blindness Cadherin Domain Cadherins Cataract Cell Adhesion Cell Culture Techniques Cell membrane Cell physiology Cellular Morphology Cellular biology Chromosome Deletion Complex Congenital chromosomal disease Cytoskeletal Modeling Data Defect Development DiGeorge Syndrome Disease Distal Electron Microscopy Ensure Epithelial Cells Etiology Event Excision F-Actin Face Generations Genes Genetic Genetic Diseases Guanosine Triphosphate Phosphohydrolases Histologic Human Intercellular Junctions Investigation Knowledge Length Lens Fiber Link Longevity MIP gene Maintenance Malignant Neoplasms Maps Measurement Mediating Microscopy Molecular Morphology Mus Mutation N-Cadherin Nature Neoplasm Metastasis Pathway interactions Penetrance Periodicity Phenotype Property Protein Dynamics Proteins Proteomics Public Health Refractive Errors Regulation Schizophrenia Shapes Shprintzen syndrome Signal Transduction Structure Techniques Testing Vision Disorders Visual impairment Water age related anticancer research biophysical properties brain research cell type cell water experimental study falls fiber cell genetic regulatory protein inhibitor lens lens transparency member microscopic imaging mutant nanocluster nervous system disorder novel preservation protein complex recruit rho GTP-Binding Proteins superresolution microscopy translational impact ultra high resolution

ARVCF gene Address Adherens Junction Adhesions Age Months Animal Model Architecture Attenuated Bilateral Binding Biochemical Biological Models Biology Biomechanics Blindness Cadherin Domain Cadherins Cataract Cell Adhesion Cell Culture Techniques Cell membrane Cell physiology Cellular Morphology Cellular biology Chromosome Deletion Complex Congenital chromosomal disease Cytoskeletal Modeling Data Defect Development DiGeorge Syndrome Disease Distal Electron Microscopy Ensure Epithelial Cells Etiology Event Excision F-Actin Face Generations Genes Genetic Genetic Diseases Guanosine Triphosphate Phosphohydrolases Histologic Human Intercellular Junctions Investigation Knowledge Length Lens Fiber Link Longevity MIP gene Maintenance Malignant Neoplasms Maps Measurement Mediating Microscopy Molecular Morphology Mus Mutation N-Cadherin Nature Neoplasm Metastasis Pathway interactions Penetrance Periodicity Phenotype Property Protein Dynamics Proteins Proteomics Public Health Refractive Errors Regulation Schizophrenia Shapes Shprintzen syndrome Signal Transduction Structure Techniques Testing Vision Disorders Visual impairment Water age related anticancer research biophysical properties brain research cell type cell water experimental study falls fiber cell genetic regulatory protein inhibitor lens lens transparency member microscopic imaging mutant nanocluster nervous system disorder novel preservation protein complex recruit rho GTP-Binding Proteins superresolution microscopy translational impact ultra high resolution

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项目摘要

Abstract: The function of the lens requires the maintenance of its transparency and refractive properties throughout its lifespan but those mechanisms responsible are not well understood. While important cellular functions and pathways such as cell adhesion or GTPase signaling have long been hypothesized to play a role in preserving these functions the removal of genes associated with these cellular functions often severely disrupt the lens making the contribution of them challenging to interpret. We have overcome this barrier with the discovery that mice lacking the Arvcf gene develop bilateral, age- dependent cortical cataracts. Arvcf is a member of the p120-catenin subfamily of catenins that bind to a specific intracellular domain of cadherins and regulate Rho-GTPases and junctional protein dynamics and stability. We have additionally found that Arvcf is highly enriched within lens fiber cell junctional structures and is required for the recruitment of several proteins to the cadherin complex, normal lens refraction, preservation of fiber cell morphology, organization of the F-actin architecture, and the normal biomechanical properties of the lens. We propose three aims to address the central question: What molecular functions lie downstream of Arvcf to maintain lens transparency, refraction, cell morphology, and biomechanical properties? In the first aim we will determine which of these properties, precede, follow, or are simultaneous with cadherin complex and adherens junction instability in Arvcf deficient lenses. We will test the hypothesis that the initiating event is the reduction of cadherin complex proteins in fiber cell membranes through a combination of biophysical property measurements of whole lenses and fluorescent/electron microscopy of lens fiber cells. Our early investigations also found that Aquaporin 0 (AQP0) depends on Arvcf to associate with the cadherin complex and novel quantitative analysis of super-resolution microscopy images of interlocking protrusions demonstrated a significant reduction of AQP0 from the distal tips of these structures. Therefore, in the second Aim of this proposal we plan to test the hypothesis that Arvcf recruits AQP0 to the tips of interlocking protrusions to facilitate cell adhesion and water transport by determining whether AQP0 and the Arvcf/N-cadherin protein complex function together in adhesion and water transport. In the third aim we will test the hypothesis that altered GTPase signaling downstream of Arvcf contributes to lens function. The GTPase regulation domain of Arvcf and candidate GTPase regulatory proteins will be genetically altered to determine their contribution to Arvcf function, lens fiber cell biology, and lens transparency. Together, these project aims will identify functional mechanisms of a previously untested and important protein required for lens transparency and lens fiber cell function. Furthermore, they will elucidate mechanisms underlying the etiology of age-dependent cortical cataracts. Because of the association of murine Arvcf with cortical cataracts and the human ARVCF gene with genetic disorders, neurological diseases, and cancer, the advances made on this proposal will also have a broad impact.

抽象的：镜头的功能需要维护其透明度和整个寿命的折射率，但那些负责的机制尚不清楚。而重要的细胞功能和途径（例如细胞）长期以来已经假设粘附或GTPase信号传导在保存这些功能中发挥作用与这些细胞功能有关解释。我们通过发现缺乏ARVCF基因的小鼠发展双侧，年龄 - 依赖性皮质白内障。 ARVCF是Catenin的P120-Catenin亚家族的成员，该亚家族与特定细胞内结合钙粘着蛋白的结构域并调节Rho-GTPase和连接蛋白动力学和稳定性。我们还发现了该ARVCF高度富集在晶状体纤维细胞连接结构中，是募集几种蛋白所必需的到钙粘蛋白复合物，正常的镜头折射，纤维细胞形态的保存，F-肌动蛋白的组织结构和镜头的正常生物力学特性。我们提出三个目标，以解决一个中心问题：什么分子函数位于ARVCF的下游，以维持晶状体透明度，折射，细胞形态和生物力学特性？在第一个目的中，我们将确定这些属性的哪个（在，遵循或同时） cadherin复合物和ARVCF缺陷透镜中的粘附连接不稳定性。我们将检验以下假设起始事件是通过生物物理的结合减少纤维细胞膜中钙粘蛋白复合物蛋白透镜纤维细胞的全镜头和荧光/电子显微镜的性质测量。我们的早期调查还发现水通道蛋白0（AQP0）依赖于ARVCF与钙粘蛋白复合物和新颖的定量相关联互锁突起的超分辨率显微镜图像的分析表明，AQP0显着降低从这些结构的远端尖端。因此，在该提议的第二个目的中，我们计划检验以下假设 ARVCF将AQP0招募到互锁突起的尖端，以促进细胞粘附和水的传输 AQP0和ARVCF/N-钙粘蛋白蛋白复合物是否在粘附和水转运中共同作用。在第三目的我们将检验以下假设，即ARVCF下游的GTPase信号传导有助于晶状体功能。 GTPase ARVCF和候选GTPase调节蛋白的调节结构域将在遗传上改变以确定其对ARVCF功能，透镜纤维细胞生物学和透镜透明度的贡献。这些项目的目的在一起将确定透镜透明度和透镜纤维细胞所需的先前未经测试和重要蛋白质的功能机制功能。此外，他们将阐明依赖年龄依赖性皮质白内障病因的机制。因为鼠ARVCF与皮质白内障和人类ARVCF基因与遗传疾病的关联疾病和癌症，该提案的进步也将产生广泛的影响。