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

• 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
• 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

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“下”运动神经元的结合变性以及“上” 大脑皮层中的运动神经元（UMNS）是ALS的重要标志。几乎所有的ALS案件都是 最终致命，疾病的快速发展使其特别可怕，超过80％ 诊断五年内死亡的患者。 ALS没有治疗，唯一可用的治疗速度很慢 疾病进展仅几个月。因此，存在更有效，更具体的需求 可以停止甚至逆转神经变性的疗法。这种疗法的创新只会由 更好地了解病理过程的分子机制，尤其是因为大多数 与ALS相关的基因普遍表达，但仅特定的细胞群体退化。 了解为什么某些单元是独特的脆弱性和绘制细胞类型特定途径的绘制 疾病期间失调的是至关重要的里程碑。这被认为是脊柱的挑战 电线投射UMN，因为它们很难与其他金字塔细胞类型区分开，因此 在临床前研究中经常被忽略。因此，他们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基于确定两个高度相似的最近工作来克服这一点 然而，在umns居住的运动皮层的5b层中，投影神经元的分子亚群。 这些人群对PON有重叠的项目，但是非重叠的项目到脊髓或 丘脑。在ALS的临床前检查这些细胞表明皮质脊髓投射神经元 （CSTN）容易受到变性的影响，而仅皮质桥骨的人群（CPN）不退化。 CSTN的选择性脆弱性可能是由于线粒体功能的失调，因为 观察到与氧化磷酸化和线索相关的基因的急剧上调。 疾病的症状阶段。本赠款的目的1将采用一种综合多摩学方法来解决 CSTN和CPN之间线粒体性质的差异是否驱动差异反应 使用一种新型的基于病毒的策略来分离疾病进展过程中细胞类型特异性线粒体的疾病 在两个临床前ALS模型中，SOD1G93A和FUSP525L。 AIM 2重点是剖析细胞的作用 鉴定出富含CSTN和细胞类型特异性生物能途径的候选基因，连接 它们具有线粒体功能和疾病脆弱性。为了提高这项工作的翻译意义， AIM 3将利用CSTN和CPN的新颖标记来进行详细的解剖学分析。 来自ALS患者的组织对通过荧光分离的细胞类型特异性核进行转录分析 验尸患者组织激活的核分子（风扇）。这项研究的结果将产生新颖 ALS中UMNS的选择性脆弱性的机制。