Normalizing E:I imbalance in Rett Syndrome by Modulation of Late Response Genes

通过调节晚期反应基因使 Rett 综合征中的 E:I 失衡正常化

基本信息

批准号：
9449084
负责人：
Rocco George Gogliotti
金额：
$ 13.64万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-19 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9449084
关键词：
Address Adopted Adverse effects Agonist Anxiety Area Atrophic Attention Attenuated Autopsy Back Behavioral Brain Brain-Derived Neurotrophic Factor CHRM4 gene Cells Cerebellum Chemosensitization Childhood Clinical Cognitive Data Disease Disease model Dose Electrophysiology (science)Etiology Excision Exhibits Exposure to Faculty Genes Genetic Glutamates Goals Head Hippocampus (Brain)Human IGF1 gene Insulin-Like Growth Factor I Intervention K-Series Research Career Programs Knock-out Knockout Mice Link Mediating Mentors Mentorship Metabotropic Glutamate Receptors Methyl-CpG-Binding Protein 2 Microcephaly Modeling Molecular Molecular Profiling Motor Motor Cortex Mus Mutation Neurodevelopmental Disorder Neurons Outcome Measure Pathology Pathway interactions Patients Pharmacology Pharmacology Study Phenotype Plant Roots Population Positioning Attribute Probability Proteins Recombinants Research Rett Syndrome Rodent Role Sampling Schizophrenia Signal Transduction Symptoms Synapses Synaptic Transmission Testing Therapeutic Tissues Training Translating career career development cholinergic neuron clinical Diagnosis conditioned fear density design experience experimental study improved loss of function mutation mouse model nervous system disorder neuron loss next generation sequencing novel positive allosteric modulator pre-clinical preference receptor relating to nervous system respiratory response social success tenure track theories tool transcription factor transcriptome sequencing treatment strategy

项目摘要

PROJECT SUMMARY AND CAREER DEVELOPMENT ABSTRACT Rett syndrome (RTT) is a devastating neurodevelopmental disorder for which there are currently no viable treatment strategies. The majority of RTT cases result from loss of function mutations in a transcription factor known as methyl CpG binding protein 2 (MECP2), and knockout of Mecp2 (Mecp2-KO) results in RTT- like phenotypes and pathologies in rodents. The brains of RTT-model mice are characterized by acquired microcephaly, which results not from neural atrophy, but rather a reduction in synapses and the subsequent simplification of the neuronal arbor. As such, the preclinical treatment strategies that have garnered the most attention are those designed to normalize synaptic density, often through the replacement of late response (LR) or direct modulation of their receptors. In this capacity RTT-mouse models are valuable tools to guide therapeutic design; however, given the challenges associated with translating data from mice to humans, we have recently adopted a different strategy. Using cortical and cerebellar tissue from 8 RTT patients, we performed RNA-sequencing (seq) analysis to screen for novel points of intervention that are rooted in human patient data, and then back-modeled relevant hits in mice. Excitingly, we identified the muscarinic acetylcholine receptor 4 (M4) as a gene significantly decreased in RTT patients. This finding is salient, as M4-modulators have been shown to rescue symptom domains in diseases like schizophrenia that overlap with RTT and M4- KO mice have cognitive and social deficits similar to Mecp2-KO mice. This raises the exciting possibility that attenuated M4 expression and signaling could be underlying cognitive and social phenotypes in RTT patients. In Aim 1, we propose to further develop preliminary data indicating that cognitive and social phenotypes in Mecp2-KO mice are responsive to M4 positive allosteric modulators (PAMs). In Aim 2, we propose to conclusively link M4-signaling to RTT in patients by perform an RNA-seq analysis on autopsy samples from patients that have been clinically diagnosed with RTT, but who do not have mutations in MECP2 (MECP2- mutation negative). Finally, in Aim 3 we propose co-administer our M4-PAM with modulators of glutamatergic and GABAergic signaling, to establish whether a complimentary symptom domains can be rescued. My career goal is to become the head of a lab with a research focus on pediatric diseases of the nervous system, and I have identified four areas of insufficient training that I propose to remedy to assist in achieving long term success in that role. The four areas are: 1) Training in electrophysiology 2) Training next generation sequencing analysis 3) Clinical exposure to RTT and 4) Mentor experience. To remove these technical and conceptual barriers, I have developed a training plan that integrates formal didactic training with hands-on mentorship, with the goal of turning these weaknesses into strengths. It is anticipated that completion of the proposed project and training plan will place me in an ideal position to receive a tenure track faculty position, and as such, I believe that this application is well suited for a K01 Career Development Award.

项目摘要和职业发展摘要雷特综合征（RTT）是一种破坏性的神经发育障碍，目前尚无治疗方法可行的治疗策略。大多数 RTT 病例是由转录功能缺失突变引起的称为甲基 CpG 结合蛋白 2 (MECP2) 的因子，敲除 Mecp2 (Mecp2-KO) 会导致 RTT- 就像啮齿动物的表型和病理学一样。 RTT 模型小鼠的大脑具有获得性特征小头畸形，这不是由神经萎缩引起的，而是由突触减少和随后的神经元乔木的简化。因此，获得最多关注的临床前治疗策略注意力是那些旨在使突触密度正常化的注意力，通常是通过替换迟发反应来实现的（LR）或直接调节其受体。在这方面，RTT 鼠标模型是指导的宝贵工具。治疗设计；然而，考虑到将小鼠数据转化为人类数据所面临的挑战，我们最近采取了不同的策略。使用 8 名 RTT 患者的皮质和小脑组织，我们进行 RNA 测序 (seq) 分析，以筛选植根于人类的新干预点患者数据，然后对小鼠的相关命中进行反向建模。令人兴奋的是，我们鉴定出了毒蕈碱乙酰胆碱 RTT 患者中受体 4 (M4) 基因显着下降。这一发现很重要，因为 M4 调节剂已被证明可以挽救精神分裂症等与 RTT 和 M4 重叠的疾病的症状域 KO 小鼠具有与 Mecp2-KO 小鼠类似的认知和社交缺陷。这提出了令人兴奋的可能性： M4 表达和信号传导减弱可能是 RTT 患者认知和社会表型的基础。在目标 1 中，我们建议进一步开发初步数据，表明认知和社会表型 Mecp2-KO 小鼠对 M4 正变构调节剂 (PAM) 有反应。在目标 2 中，我们建议通过对尸检样本进行 RNA 序列分析，最终将 M4 信号传导与患者的 RTT 联系起来已被临床诊断为 RTT 但 MECP2 没有突变的患者（MECP2- 突变阴性）。最后，在目标 3 中，我们建议将 M4-PAM 与谷氨酸能调节剂共同给药和 GABAergic 信号传导，以确定是否可以挽救互补的症状域。我的职业目标是成为一个专注于儿科疾病研究的实验室负责人神经系统，我已经确定了四个训练不足的领域，我建议对其进行补救，以帮助在此职位上取得长期成功。这四个领域是： 1) 电生理学培训 2) 接下来的培训世代测序分析 3) 临床接触 RTT 和 4) 导师经验。要删除这些由于技术和概念上的障碍，我制定了一个培训计划，将正式的教学培训与实践指导，旨在将这些弱点转化为优势。预计完成拟议的项目和培训计划将使我处于获得终身教职的理想位置职位，因此，我相信这个应用程序非常适合 K01 职业发展奖。