YAP/TAZ in Schwann Cells as Potential Therapeutic Targets in CMT1A and HNPP.

雪旺细胞中的 YAP/TAZ 作为 CMT1A 和 HNPP 的潜在治疗靶点。

• 批准号: 10730544
• 负责人: Seth Michael Moore
• 金额: $ 3.33万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10730544
• 关键词: Ablation; Age Months; Alleles; Animal Model; Biochemical; Biopsy; Biopsy Specimen; Cell Cycle; Cell Cycle Regulation; Cell Differentiation process; Cell Nucleus; Cell Proliferation; Cellular biology; Charcot-Marie-Tooth Disease; Clinic; Clinical; Communication; Complex; Critical Thinking; Cryoultramicrotomy; Cytoskeleton; Dedications; Defect; Demyelinations; Development; Disease; EGR2 gene; Elasticity; Electrons; Electrophysiology (science); Embryo; Environment; GTP-Binding Proteins; Gene Deletion; Genes; Genetic; Genetic Transcription; Glutaminase; Goals; Hereditary neuropathy with liability to pressure palsies; Heterozygote; Hospitals; Human; Immunohistochemistry; Inherited; Integrins; Iowa; Italy; Knock-out; Laminin Receptor; Lipids; Maintenance; Mediating; Microscopic; Modeling; Morphology; Mus; Myelin; Nerve; Nerve Fibers; Neuroglia; Neuropathy; Nuclear; PMP22 gene; Pathologic; Pathology; Pathway interactions; Patients; Peripheral Nerves; Peripheral Nervous System Diseases; Permeability; Phenotype; Phosphorylation; Phosphotransferases; Physicians; Physiology; Prevalence; Problem Solving; Proliferating; Proteins; Radial; Regulation; Research; Resistance; Role; Sampling; Schwann Cells; Scientist; Sensory; Signal Pathway; Skin; Sorting; Source; Tamoxifen; Techniques; Testing; Therapeutic; Thinness; Time; Tooth Diseases; Training; Transcription Coactivator; Transcriptional Coactivator with PDZ-Binding Motif; Transcriptional Regulation; Tumor Suppressor Proteins; Universities; Western Blotting; behavioral phenotyping; career; conditional knockout; congenic; disease phenotype; dosage; experience; experimental study; improved; mRNA Expression; mechanical properties; mechanical signal; mechanical stimulus; mouse model; myelination; neurophysiology; novel; novel therapeutics; overexpression; postnatal; receptor; response; sciatic nerve; skills; sural nerve; therapeutic target; transcription factor; transmission process

Project Summary The heterogeneous group of inherited peripheral neuropathies referred to as Charcot-Marie Tooth disease, have an estimated prevalence of 1:25001. Among these disorders, Charcot Marie Tooth 1A (CMT1A) and Hereditary Neuropathy with Liability to Pressure Palsy (HNPP) are the most common1. These diseases result from duplication (CMT1A) or deletion (HNPP) of the gene encoding for peripheral myelin protein 22 (PMP22)2, leading to its over or underexpression in Schwann cells (SCs), respectively3,4. There are no cures for these debilitating diseases, highlighting the need to identify novel mechanisms to modulate PMP22 expression, and to elucidate the pathological mechanisms involved in peripheral nerve (PN) demyelination. Recently, the complex formed by the transcriptional coactivator with PDZ-binding motif (TAZ)5 and the transcription factor TEA domain 1 (TEAD1)6,7, was shown to positively regulate PMP22 expression8,9. Therefore, modulation of TAZ activation is an intriguing therapeutic avenue to decrease PMP22 transcription in CMT1A, or increase its transcription in HNPP. Moreover, the functions of PMP22 in SC biology include cell cycle regulation10,11, formation of cellular junctions and myelin permeability12,13, and determining the mechanical properties of myelinated PN fibers14,15. Notably, these aspects of SC biology are also regulated by TAZ, and the related transcriptional coactivator Yes- associated protein 1 (YAP)16–20; the major downstream effectors of the HIPPO pathway21,22. Activated YAP/TAZ translocate to the nucleus and associate with TEAD1-423,24, altering the transcription of many genes essential for SC proliferation and differentiation including laminin receptors, integrins, G-protein Ga, EGR2, and myelin and lipid genes8,20,25,26. Therefore, changes in the activation status of YAP/TAZ in CMT1A and HNPP may impact the physiology of SCs, contributing to demyelination. Thus, we hypothesize that modifying TAZ expression may restore proper PMP22 expression and ameliorate the phenotypes of CMT1A through direct regulation of PMP22. We also postulate that YAP/TAZ activation may be altered in mouse models and human samples of CMT1A and HNPP, potentially contributing to altered SC proliferation, myelin permeability, and mechanical properties of PNs. To determine the effects of modulating TAZ activation, genetic ablation of TAZ alleles in SCs of PMP22- overexpressing mice will be assessed for morphological, electrophysiological, sensory, and behavioral phenotypic rescue. The presence of altered YAP/TAZ activation will be assessed through immunostaining and biochemical analyses of human biopsy samples and mouse models of CMT1A and HNPP. The research for this proposed project will be conducted at the Institute for Myelin and Glia Exploration, composed of an interdisciplinary group of scientists dedicated to the study of myelin and its associated diseases. The overall training plan will emphasize the development of scientific communication skills, training in various techniques, improving critical thinking and problem-solving skills, as well as opportunities for regular clinical experiences. This will create the ideal environment for developing the skills necessary for a career as a physician-scientist.

项目摘要 遗传性周围神经病的异质性组被称为Charcot-Marie Tooth病， 估计患病率为1：25001。在这些疾病中，Charcot玛丽牙1A（CMT 1A）和遗传性 神经病变与压力麻痹（HNPP）的责任是最常见的1。这些疾病是由 编码外周髓磷脂蛋白22（PMP 22）2的基因的重复（CMT 1A）或缺失（HNPP），导致 其在雪旺细胞（SC）中的过度表达或表达不足，分别为3，4。这些使人衰弱的疾病 疾病，强调需要确定新的机制来调节PMP 22的表达，并阐明 周围神经脱髓鞘的病理机制。最近， 具有PDZ结合基序（TAZ）5和转录因子TEA结构域1的转录共激活因子 （TEAD 1）6，7显示出正调节PMP 22表达8，9。因此，TAZ激活的调节是 一个有趣的治疗途径，以减少PMP 22在CMT 1A中的转录，或增加其在CMT 1A中的转录。 HNPP。此外，PMP 22在SC生物学中的功能包括细胞周期调节10，11，细胞周期的形成，以及细胞内的细胞凋亡。 连接和髓鞘渗透性12，13，并确定有髓鞘PN纤维的机械性能14，15。 值得注意的是，SC生物学的这些方面也受到TAZ和相关转录辅激活因子的调节。 相关蛋白1（雅普）16-20; HIPPO途径的主要下游效应物21，22。激活的雅普/TAZ 转移到细胞核并与TEAD 1 - 423，24相关，改变许多必需基因的转录 包括层粘连蛋白受体、整联蛋白、G蛋白Ga、EGFR 2和髓磷脂 和脂质基因8，20，25，26。因此，CMT 1A和HNPP中雅普/TAZ激活状态的变化可能会影响 SC的生理学，有助于脱髓鞘。因此，我们假设，改变TAZ表达可能 通过直接调节PMP 22恢复PMP 22的正常表达并改善CMT 1A的表型。 我们还假设雅普/TAZ激活可能在小鼠模型和CMT 1A的人样品中改变， HNPP，可能有助于改变SC增殖，髓鞘渗透性和PN的机械性能。 为了确定调节TAZ激活的作用，在PMP 22 - 24的SC中遗传消除TAZ等位基因。 将评估过表达小鼠的形态学、电生理学、感觉和行为 表型拯救将通过免疫染色评估是否存在改变的雅普/TAZ激活 CMT 1A和HNPP的人活检样品和小鼠模型的生物化学分析。混输泵的研究 拟议的项目将在髓鞘和神经胶质探索研究所进行，由一个 致力于髓鞘及其相关疾病研究的跨学科科学家小组。整体 培训计划将强调发展科学交流技能，培训各种技术， 提高批判性思维和解决问题的能力，以及定期临床经验的机会。 这将为培养作为一名医生-科学家的职业所需的技能创造理想的环境。