Function and Regulation of PMP22 in CMT1A and HNPP

PMP22 在 CMT1A 和 HNPP 中的功能和调控

• 批准号: 10350403
• 负责人: Kathryn Renae Moss
• 金额: $ 12.3万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-21 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10350403
• 关键词: Adherens Junction; Adhesions; Affect; Aging; Award; Axon; Biophysics; Canis familiaris; Career Transition Award; Cell Adhesion; Cell membrane; Cell model; Cells; Charcot-Marie-Tooth Disease; Complement; Complex; Confocal Microscopy; Cultured Cells; Development; Disease; Electrical Resistance; Electron Microscopy; Gene Dosage; Genes; Goals; Hereditary neuropathy with liability to pressure palsies; Human; Inherited; Intercellular Junctions; Investigation; Knowledge; Label; MDCK cell; Membrane; Membrane Microdomains; Mentorship; Microscopy; Modeling; Modification; Molecular Neurobiology; Morphology; Myelin; Nerve; Nerve Fibers; PMP22 gene; Pathogenesis; Peripheral Nerves; Peripheral Nervous System Diseases; Phase; Phenotype; Positioning Attribute; Regulation; Research; Research Personnel; Resolution; Role; Schwann Cells; Tamoxifen; Technical Expertise; Techniques; Therapeutic; Tight Junctions; Training; Vesicle; Work; biophysical properties; biophysical techniques; career; confocal imaging; dosage; dysmyelination; improved; kidney epithelial cell; mouse model; mutant; myelination; overexpression; palmitoylation; protein function; recombinase-mediated cassette exchange; success; therapeutically effective; therapy development

Project Summary/Abstract The fact that both duplication and deletion of the Peripheral Myelin Protein 22 (PMP22) gene cause dysmyelinating peripheral neuropathy illustrates the importance of PMP22 for peripheral myelin integrity. PMP22 duplication causes Charcot-Marie-Tooth Disease Type 1A (CMT1A) and PMP22 deletion causes Hereditary Neuropathy with Liability to Pressure Palsies (HNPP). Although CMT1A and HNPP are the most common inherited peripheral neuropathies, research on them is underfunded and there are currently no disease-modifying treatments. This is largely due to the fact that PMP22 function and the consequence of altered PMP22 expression remain unclear. Thus, there is a critical need to expand knowledge of what PMP22 does in myelin, how it is regulated and when precise expression is required as a means to improve therapeutic potential of CMT1A and HNPP. This proposal aims to utilize cutting-edge techniques, including conditional mouse models and super resolution microscopy, and knowledge of cell adhesion and membrane biophysics to advance understanding of CMT1A and HNPP pathomechanisms. My central hypothesis is that PMP22 gene dosage and lipid raft association govern localization and organization of myelin adherens and tight junctions; a function that is most critical during development. In Aim 1, I will determine how PMP22 regulates adhesion junction organization in peripheral nerve myelin during development and aging with super resolution and electron microscopy. I will aid the interpretation of these studies by evaluating the effects of altered PMP22 expression on prototypical adherens and tight junctions in Madin-Darby Canine Kidney (MDCK) epithelial cells. The temporal requirement for precise PMP22 expression in myelin will also be defined by generating powerful conditional mouse models of CMT1A and HNPP. In Aim 2, I will determine how palmitoylation impacts PMP22 lipid raft association and regulation of adhesion junctions and define the biophysical properties of PMP22 within the plasma membrane using MDCK and Schwann cell models of CMT1A and HNPP and advanced biophysical methods. This K22 Career Transition Award will provide me with additional training, mentorship and expertise in cell adhesion, membrane biophysics and microscopy, thereby enabling my proposed research and facilitating my transition to independence. This training will complement my previous training in cell and molecular neurobiology and my current peripheral nerve training, and the expertise acquired during Phase I will be applied to more complex models in Phase II to expand mechanistic details. Completion of these aims will accelerate progress towards my long-term goal of developing an independent academic research career studying pathomechanisms of CMT1A and HNPP as a means to improve their therapeutic potential. The training and mentorship provided by this award will expand my technical skills and expertise, positioning me for success as an independent investigator.

项目摘要/摘要 外周髓磷脂蛋白22（PMP22）基因的重复和缺失的事实 肿瘤的外周神经病说明了PMP22对周围髓磷脂完整性的重要性。 PMP22复制导致charcot-marie-tooth疾病1A型（CMT1A）和PMP22缺失原因 遗传性神经病对麻痹（HNPP）负有责任。虽然CMT1A和HNPP是最大的 常见的遗传外周神经病，对它们的研究资金不足，目前没有 疾病改良治疗。这主要是由于PMP22功能和结果 PMP22表达的改变尚不清楚。因此，迫切需要扩大了解PMP22的知识 在髓磷脂中进行调节，以及需要精确表达以作为改善治疗的一种手段 CMT1A和HNPP的潜力。该建议旨在利用尖端技术，包括条件 小鼠模型和超级分辨率显微镜，以及细胞粘附和膜生物物理学的知识 提前了解CMT1A和HNPP致病机制。我的中心假设是PMP22基因 剂量和脂质筏协会管理髓磷脂膜的本地化和组织 连接；在开发过程中最关键的功能。在AIM 1中，我将确定PMP22如何 调节外周神经髓磷脂的粘附连接组织在开发和衰老中 分辨率和电子显微镜。我将通过评估这些研究的解释 Madin-Darby犬肾（MDCK）的原型粘附剂和紧密连接的PMP22表达改变了 上皮细胞。髓磷脂中精确PMP22表达的时间要求也将由 生成CMT1A和HNPP的强大条件鼠标模型。在AIM 2中，我将确定如何 棕榈酰化影响PMP22脂质筏关联和粘附连接处的调节，并定义 使用MDCK和Schwann细胞模型的PMP22的生物物理特性 CMT1A和HNPP以及晚期生物物理方法。这个K22职业过渡奖将为我提供 细胞粘附，膜生物物理学和显微镜的其他培训，指导和专业知识，从而 使我提出的研究并促进我向独立的过渡。该培训将补充 我以前在细胞和分子神经生物学和目前的外周神经训练中训练，以及 在第一阶段获得的专业知识将应用于II阶段更复杂的模型以扩展机理 细节。这些目标的完成将加速我的长期目标的进步 独立的学术研究职业研究CMT1A和HNPP的病理机理作为一种手段 提高其治疗潜力。该奖项提供的培训和指导将扩大我的 技术技能和专业知识，使我成为独立调查员的成功。