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：
10745149
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：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10745149
关键词：
ALS pathology ALS patients Address Affinity Chromatography Anatomy Animal Model Antioxidants Autopsy Biochemical Biochemistry Bioenergetics Brain Stem Candidate Disease Gene Cell Nucleus Cell Separation Cells Cerebral cortex Clinical Corticospinal Tracts Data Diagnosis Disease Disease Progression Engineering Etiology Fluorescence Functional disorder Gene Expression Gene Expression Profiling Gene Expression Regulation Genes Goals Grant Innovative Therapy Intervention Label Link Maps Measures Metabolic Mitochondria Modeling Molecular Molecular Profiling Motor Cortex Motor Neuron Disease Motor Neurons Mus Mutation Nerve Degeneration Nervous System Neurons Nuclear Onset of illness Oxidative Phosphorylation Oxidative Stress Pathologic Processes Pathology Pathway interactions Patients Pontine structure Population Pre-Clinical Model Predisposition Property Proteins Proteomics Pyramidal Cells Pyramidal Tracts RNA-Binding Protein FUS Research Rest Ribosomes Role Sorting Spinal Cord Symptoms Techniques Testing Thalamic structure Tissues Transcript Transgenic Mice Translating Up-Regulation Viral Viral Vector Vulnerable Populations Work biological adaptation to stress biomarker identification candidate identification cell cortex cell type cellular resilience comparative complex IV disease-causing mutation end stage disease gain of function human tissue innovation loss of function metabolomics molecular phenotype multiple omics neuropathology new therapeutic target novel novel marker pre-clinical preclinical study programs resilience response translational framework

项目摘要

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, especially since 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. This has posed a considerable challenge for the spinal cord projecting 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 ALS-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 dissecting the cellular role of identified candidate genes that are enriched in CSTNs and cell type-specific bioenergetic pathways, linking them to mitochondrial function and disease vulnerability. 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 to 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.

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。更好地了解病理过程背后的分子机制，特别是因为大多数与 ALS 相关的基因普遍表达，但只有特定的细胞群会退化。了解为什么某些细胞特别脆弱，并绘制细胞类型特定的途径疾病期间的失调是重要的里程碑，这对脊髓提出了相当大的挑战。脐带突出 UMN，因为它们很难与其他锥体细胞类型区分开，因此因此，它们在临床前研究中经常被忽视，这是导致 ALS 的选择性脆弱性的基础。突变仍然是一个谜。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，Aim 2 重点剖析 ALS 的细胞作用。确定了富含 CSTN 和细胞类型特异性生物能途径的候选基因，将它们对线粒体功能和疾病脆弱性的影响增加了这项工作的转化意义，目标 3 将利用 CSTN 和 CPN 的新型标记物对尸检进行详细的解剖分析来自 ALS 患者的组织，对通过荧光分离的细胞类型特异性细胞核进行转录分析这项研究的结果将产生新颖的结果。 ALS 中 UMN 选择性脆弱性的机制。