Molecular Phenotyping of Cortical Cell Types in ALS Related Neurodegeneration

ALS 相关神经变性中皮质细胞类型的分子表型

基本信息

批准号：
10592732
负责人：
Eric F Schmidt
金额：
$ 42.38万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10592732
关键词：
ALS pathology ALS patients Address Affinity Chromatography Anatomy Animal Model Autopsy Biochemical Biochemistry Brain Stem Candidate Disease Gene Cell Nucleus Cells Cerebral cortex Clinical Corticospinal Tracts Data Diagnosis Disease Disease Progression Engineering Etiology Fluorescence Functional disorder Funding Future Gene Expression Gene Expression Profiling Genes Goals Grant Innovative Therapy Intervention Label Lead Link Measures Metabolic Mitochondria Modeling Molecular Molecular Profiling Morphology Motor Cortex Motor Neuron Disease Motor Neurons Mus Nerve Degeneration Nervous system structure Neurodegenerative Disorders Neurons Nuclear Onset of illness Oxidative Phosphorylation Pathologic Processes Pathology Pathway interactions Patients Pontine structure Population Pre-Clinical Model Predisposition Property Proteins Proteomics Pyramidal Cells Research Rest Ribosomes Role Sorting - Cell Movement Spinal Cord Symptoms Techniques Testing Thalamic structure Tissues Transcript Transgenic Mice Translating Up-Regulation Viral Viral Vector Vulnerable Populations Work base biological adaptation to stress cell type comparative disease-causing mutation experimental study familial amyotrophic lateral sclerosis human tissue innovation loss of function metabolic profile metabolomics molecular phenotype motor neuron degeneration multiple omics neuropathology new therapeutic target novel novel marker overexpression pre-clinical preclinical study programs response

项目摘要

SUMMARY The combined degeneration of both “lower” motor neurons in the brainstem and spinal cord and “upper” motor neurons (UMNs) in the cerebral cortex is an important hallmark of ALS. Almost all cases of ALS are eventually fatal, and the rapid progression of the disease makes it particularly terrible, with over 80% of patients dying within five years of diagnosis. No cure exists for ALS and the only available treatments slow disease progression by merely a few months. Therefore, a great need exists for more effective and specific therapies that can stop or even reverse neurodegeneration. Innovation for such therapies will only arise from a better understanding of the molecular mechanisms underlying the pathological process. Most genes linked to ALS are ubiquitously expressed yet only specific populations of cells degenerate. Understanding why certain cells are uniquely vulnerable and mapping cell type specific pathways that are dysregulated during disease are crucial milestones for developing innovative therapies. This has posed a considerable challenge for the spinal cord-projecting upper motor neurons (UMNs) since they are difficult to distinguish from other pyramidal cell types and are therefore often overlooked in preclinical studies. Because of this, the basis for their selective vulnerability to disease-causing mutations has remained a mystery. The proposed study aims to overcome this by building on recent work that identified two highly similar yet molecularly distinct subpopulations of projection neurons in layer 5b of motor cortex, where UMNs reside. These populations have overlapping projections to pons, but non-overlapping projections to the spinal cord or thalamus. Examining these cells in preclinical models of ALS revealed that the corticospinal projecting neurons (CSTNs) were vulnerable to degeneration, while the corticopontine-only population (CPN) did not degenerate. The selective vulnerability of the CSTNs was likely due to dysregulation of mitochondrial function since a dramatic upregulation of genes related to oxidative phosphorylation and mitophagy was observed at symptomatic stages of disease. Aim 1 of this grant will employ an integrative multi-omics approach to address whether differences in the properties of mitochondria between CSTNs and CPNs drive differential responses to disease using a novel, viral-based strategy to isolate cell type specific mitochondria during disease progression in two preclinical ALS models, SOD1G93A and FUSP525L. Aim 2 focuses on characterizing the cellular role of identified candidate genes that are enriched in CSTNs, shown to localize to mitochondria, and have been directly linked to clinical cases of ALS and other neurodegenerative disorders. To increase the translational significance of this work, Aim 3 will leverage novel markers for CSTNs and CPNs for a detailed anatomical analysis of postmortem tissue from ALS patients perform transcriptional profiling on cell type specific nuclei isolated by fluorescence activated nuclear sorting (FANS) from postmortem patient tissue. Results from this study will yield novel mechanisms underlying selective vulnerability of UMNs in ALS.

概括脑干和脊髓中的“下”运动神经元和“上”运动神经元的联合变性大脑皮层的运动神经元 (UMN) 是 ALS 的重要标志，几乎所有 ALS 病例都是如此。最终致命，而且疾病的快速进展使其变得尤为可怕，超过80% 患者在诊断后五年内死亡，目前尚无治愈 ALS 的方法，唯一可用的治疗方法缓慢。因此，非常需要更有效和特异性的治疗。能够阻止甚至逆转神经退行性变的疗法只能来自于创新。更好地了解与病理过程相关的大多数基因的分子机制。 ALS 普遍表达，但只有特定细胞群会退化。细胞特别脆弱，绘制疾病期间失调的细胞类型特异性途径是开发创新疗法的关键里程碑这对脊柱提出了相当大的挑战。脊髓投射上运动神经元 (UMN)，因为它们很难与其他锥体细胞区分开来类型，因此在临床前研究中经常被忽视，因此，它们是选择性的基础。致病突变的脆弱性仍然是一个谜。拟议的研究旨在通过最近的工作来克服这个问题，该工作确定了两个高度相似的然而，UMN 所在的运动皮层 5b 层中的投射神经元亚群在分子上却截然不同。这些人群对脑桥有重叠的投影，但对脊髓或脊髓的投影不重叠在 ALS 临床前模型中检查这些细胞发现，皮质脊髓投射神经元（CSTN）很容易退化，而仅皮质桥蛋白群体（CPN）则不会退化。 CSTN 的选择性脆弱性可能是由于自线粒体功能失调以来观察到与氧化磷酸化和线粒体自噬相关的基因急剧上调该赠款的目标 1 将采用综合多组学方法来解决疾病的症状阶段。 CSTN 和 CPN 之间线粒体特性的差异是否会导致不同的反应使用基于病毒的新型策略在疾病进展过程中分离细胞类型特异性线粒体在两个临床前 ALS 模型 SOD1G93A 和 FUSP525L 中，目标 2 侧重于表征 ALS 的细胞作用。鉴定出富含 CSTN 的候选基因，显示其定位于线粒体，并且已被与 ALS 和其他神经退行性疾病的临床病例直接相关为了体现这项工作的重要性，Aim 3 将利用 CSTN 和 CPN 的新颖标记来进行详细的解剖学研究对 ALS 患者死后组织进行分析，对细胞类型特异性细胞核进行转录分析通过荧光激活核分选（FANS）从死后患者组织中分离得到此结果。研究将产生 ALS 中 UMN 选择性脆弱性的新机制。