Mouse models of ATP1A1 mutation -linked neuropathies

ATP1A1 突变相关神经病的小鼠模型

基本信息

批准号：
10093169
负责人：
Pablo Artigas
金额：
$ 7.65万
依托单位：
TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10093169
关键词：
ATP1A1 gene Address Adult Age Age of Onset Alleles Animal Model Axon Behavioral Biological Models Cell membrane Cells Characteristics Charcot-Marie-Tooth Disease Cognitive Communities Complement Data Development Disease Evaluation Exercise Family Fiber Functional disorder Future Generations Genes Genetic Transcription Goals Hereditary Motor and Sensory-Neuropathy Type II Histologic Human Human body Hyperaldosteronism Hypomagnesemia In Vitro Induced Mutation Isoenzymes Knock-out Knockout Mice Laboratories Light Link Longevity LoxP-flanked allele Masks Modeling Motor Mus Mutation Myocardium Na(+)-K(+)-Exchanging ATPase Nervous system structure Neuraxis Neurofilament Proteins Neurologic Neurons Neuropathy Patients Peripheral Peripheral Nervous System Diseases Phenotype Physiological Physiology Primary Hyperaldosteronism Protein Isoforms Proteins RNA Role Secondary Hypertension Seizures Sensory Severities Skeletal Muscle Symptoms Tamoxifen Techniques Technology Testing Time Tissues age related axonal degeneration conditional knockout experimental study insight loss of function loss of function mutation mouse model mutant novel prevent protein expression tool

项目摘要

Charcot-Marie-Tooth (CMT) comprises a heterogeneous group of peripheral neuropathies caused by mutations in over 90 genes. Mutation of ATP1A1, which encodes for the Na+,K+-ATPase (NKA) α1 subunit, has been recently associated with CMT2, a CMT form characterized by axonal degeneration. NKA is a heterodimeric (αβ) protein that hydrolyzes ATP to build and maintain the Na+ and K+ gradients across the plasma membrane of all human cells. Different tissues present distinct NKA isozymes. ATP1A1 is ubiquitously expressed and the mutations linked to CMT2 cause NKA loss of function. Loss of NKA function is also observed in ATP1A1 mutants associated with other diseases, including primary hyperaldosteronism and a form of hypomagnesemia accompanied by seizures and cognitive delay. However, the lack of appropriate model systems has prevented a detailed understanding of the pathophysiology of these ATP1A1 mutation linked disorders. The long-term goal of our laboratory is to understand the mechanisms of NKA function and their roles in physiological and disease states. The objective of this proposal is to develop and evaluate animal models to study ATP1A1-linked disease, with an emphasis on CMT2. Aim 1. Comprehensive neuropathic evaluation of heterozygous ATP1A1 knockout mice to test our central hypothesis, that the severe effect of loss-of-function mutations seen in CMT2 patients, including the highly variables symptom intensity and age of onset, should be recapitulated in ATP1A1+/- mice. Aim 2. Develop and evaluate novel ATP1A1 loss-of-function-mutation models using CRE-LoxP technology to test the hypotheses that deletion of one ATP1A1 allele in adulthood accelerates the onset of CMT2 symptoms and that neuronal haploinsufficiency is sufficient to induce CMT2. Through the generation of tissue- and time-dependent conditional knockout mice using tamoxifen inducible CreER lines, we will be able to determine age-dependent compensatory mechanisms, and necessity of local or systemic haploinsufficiency for CMT2 induction. Successful completion of these studies will lead to viable NKA-linked pathophysiological model to gain insight into CMT2 mechanisms. The results from this project will provide a tool for future testing of specific treatments for NKA-linked CMT2. Additionally, the experiments outlined here are likely to provide models for the other ATP1A1 mutation-linked diseases. Scientifically, our results will uncover the functional roles of NKA α1 in neuron physiology elucidating its importance in both the peripheral and central nervous systems. These mouse models will be made available to the scientific community through standard multi-institutional MTAs.

腓骨肌萎缩症 (CMT) 包括由以下原因引起的一组异质性周围神经病：编码 Na+,K+-ATP 酶 (NKA) α1 亚基的 ATP1A1 发生突变。最近与 CMT2 相关，NKA 是一种以轴突变性为特征的 CMT 形式，它是一种异二聚体 (αβ) 蛋白，可水解 ATP 以在所有人类细胞的质膜上建立和维持 Na+ 和 K+ 梯度。 ATP1A1 普遍表达，与 CMT2 相关的突变导致 NKA 功能丧失。在 ATP1A1 突变体中也观察到 NKA 功能丧失。然而，缺乏适当的模型系统阻碍了对这些 ATP1A1 突变相关疾病的病理生理学的详细了解。了解 NKA 功能的机制及其在生理和疾病状态中的作用该提案的目的是开发和评估动物模型以研究 ATP1A1 相关疾病，重点是 CMT2。 1. 对高度杂合的 ATP1A1 敲除小鼠进行全面的神经病理评估，以测试我们的中心假设，即 CMT2 患者中观察到的功能丧失突变的严重影响，包括可变的症状强度和发病年龄，应在 ATP1A1+/- 中重述目标 2. 使用 CRE-LoxP 技术开发和评估新型 ATP1A1 功能丧失突变模型，以测试删除一个的假设。成年期的 ATP1A1 等位基因会加速 CMT2 症状的发生，并且神经单倍体不足足以诱导 CMT2。通过使用他莫昔芬诱导的 CreER 系生成组织和时间依赖性条件敲除小鼠，我们将能够确定年龄依赖性补偿机制。，以及 CMT2 诱导局部或全身单倍体不足的必要性，成功完成这些研究将产生可行的 NKA 相关病理生理学模型，以获得深入的了解。该项目的结果将为未来测试 NKA 相关 CMT2 的特定治疗提供工具，从科学上讲，我们的结果可能会为其他 ATP1A1 突变相关疾病提供模型。揭示 NKA α1 在神经元生理学中的功能作用，阐明其在外周和中枢神经系统中的重要性。这些小鼠模型将通过标准的多机构 MTA 提供给科学界。