Use of IPSC to define role of astrocytes in specifying risk for onset of cerebral adrenoleukodystrophy

使用 IPSC 来定义星形胶质细胞在确定脑肾上腺脑白质营养不良发作风险中的作用

• 批准号: 10118513
• 负责人: Jaspreet Singh
• 金额: $ 37.63万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10118513
• 关键词: Adrenoleukodystrophy; Adrenomyeloneuropathy; Allogenic; Area; Astrocytes; Autologous; Automobile Driving; Autopsy; Biochemical; Body Fluids; Brain; Brain Diseases; CRISPR/Cas technology; Carbon; Cells; Cerebrum; Citric Acid Cycle; Clinical; Clinical Course of Disease; Data; Defect; Deletion Mutation; Development; Diagnosis; Disease; Disease Progression; Distant; Drug Screening; Electron Transport; Energy Metabolism; Environment; Exposure to; Eye; Female; Fibroblasts; Genes; Genetic Diseases; Genotype; Glucose; Glycolysis; Goals; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Immune; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Inherited; Investigation; Knock-in; Laboratories; Laboratory Study; Lead; Life; Measures; Mediating; Membrane Proteins; Metabolic; Metabolic Pathway; Mitochondria; Modeling; Molecular; Mutation; Nature; Neonatal Screening; Neurologic; Newly Diagnosed; Pathway interactions; Patients; Phenotype; Plasma; Play; Procedures; Protein Import; Public Health; Reporting; Research; Respiration; Risk; Role; Siblings; Source; Specific qualifier value; Spinal Cord Diseases; Stem Cell Research; Stimulus; Supplementation; Testing; Time; Tissues; Translating; Up-Regulation; Very Long Chain Fatty Acid; axonopathy; clinical predictors; cytokine; drug development; effective therapy; extracellular; gene therapy; improved; in vivo; induced pluripotent stem cell; innovation; male; metabolomics; mitochondrial dysfunction; motor disorder; mouse model; neuroinflammation; novel; peroxisome; response; screening panel; screening program; transcriptome; white matter

PROJECT SUMMARY/ABSTRACT The mechanism of onset of neuroinflammation in fatal phenotypes in males with inherited X-linked adrenoleukodystrophy (X-ALD) disease remains unknown. 60% of male X-ALD patients develop fatal cerebral neuroinflammation (cALD) while remaining develop milder adrenomyeloneuropathy (AMN) characterized by axonopathy without neuroinflammation. The primary genetic defect in X-ALD (mutation/deletion in ABCD1 gene) and the biochemical defect (accumulation of very long chain fatty acid; C>22:0 in plasma and tissues) cannot predict the onset of AMN or cALD. Our long-term goal is to dissect the molecular mechanism underlying differential phenotype development in X-ALD. The objective of this application is to identify metabolic pathways that underlie the differential neuroinflammatory response in AMN and cALD human astrocytes. These astrocytes were differentiated from induced pluripotent stem cells (iPSCs), which in turn were generated by reprogramming of human control, AMN and cALD patient-derived fibroblasts. Metabolic reprogramming is emerging as a novel regulator of inflammatory response. Astrocytes rely on mitochondrial respiration (OXPHOS) for their metabolic needs but switch to glycolysis under neuroinflammatory environment to boost biosynthetic pathways to produce inflammatory mediators. Our preliminary proof-of-concept data, with untargeted metabolomics, identified metabolites altered between healthy-control and cALD phenotype postmortem brain. Within the cALD brain white matter, unique metabolite changes were recorded between distant normal looking areas and areas adjacent to the plaque suggesting an association with disease progression. We found both OXPHOS and glycolysis decreased (low metabolic state) in human cALD astrocytes despite higher inflammatory response. This low metabolic state suggests role of novel alternative source(s) of fuel driving the neuroinflammatory response in cALD astrocytes. Our central hypothesis is that metabolic reprogramming in cALD astrocytes drives their proinflammatory shift that underlies the neuroinflammatory disease progression in cALD. To test our hypothesis we propose two specific aims: 1) To elucidate the metabolic reprogramming responsible for inflammatory response in cALD astrocytes. 2) To determine if dysfunctional mitochondria play a role in inflammatory nature of cALD astrocytes? We will take advantage of control, AMN and cALD astrocytes generated from iPSC’s in our laboratory for these studies. This proposal is innovative, because it departs from the status quo by identifying for the first time, metabolic pathways differentially regulating inflammatory response in human AMN and cALD astrocytes. The proposed research is significant because the cellular mechanism(s) that lead to less severe AMN or fatal cALD phenotype in response to same ABCD1 mutation remain unknown even four decades after the identification of gene defect in X-ALD. Impact: With the rising rate of newly diagnosed cases after X-ALD was added to the federal newborn screening list in 2016, there is urgent need to identify novel targets to develop effective therapies for AMN and cALD for which no satisfactory therapy exists.

项目概要/摘要 X连锁遗传性男性致命表型中神经炎症的发病机制 肾上腺脑白质营养不良 (X-ALD) 疾病仍不清楚，60% 的男性 X-ALD 患者会出现致命性脑病。 神经炎症 (cALD)，同时仍发展为轻度肾上腺脊髓神经病 (AMN)，其特征为 无神经炎症的轴突病。X-ALD 的主要遗传缺陷（ABCD1 基因突变/缺失）。 生化缺陷（极长链脂肪酸的积累；血浆和组织中C>22:0）不能 预测 AMN 或 cALD 的发生 我们的长期目标是剖析潜在的分子机制。 X-ALD 中的差异表型发育 本应用的目的是确定代谢途径。 这是 AMN 和 cALD 人类星形胶质细胞中不同神经炎症反应的基础。 是从诱导多能干细胞（iPSC）分化而来，而诱导多能干细胞又是通过重新编程产生的 人类对照、AMN 和 cALD 患者来源的成纤维细胞的代谢重编程正在成为一种新型药物。 星形胶质细胞的代谢依赖于线粒体呼吸（OXPHOS）。 需要但在神经炎症环境下转向糖酵解以促进生物合成途径产生 我们通过非靶向代谢组学的初步概念验证数据确定了炎症介质。 在 cALD 脑白质中，健康对照和 cALD 表型之间的代谢物发生了变化。 物质，在遥远的正常区域和邻近区域之间记录了独特的代谢变化 我们发现 OXPHOS 和糖酵解都与斑块有关。 尽管炎症反应较高，但人类 cALD 星形胶质细胞的代谢水平降低（低代谢状态）。 代谢状态表明新的替代燃料来源在驱动神经炎症反应中的作用 我们的中心假设是 cALD 星形胶质细胞的代谢重编程驱动它们的生长。 促炎症转变是 cALD 神经炎症疾病进展的基础，以检验我们的假设。 我们提出两个具体目标：1）阐明导致炎症的代谢重编程 cALD 星形胶质细胞的反应 2) 确定功能失调的线粒体是否在炎症中发挥作用。 cALD 星形胶质细胞？我们将利用 iPSC 生成的对照、AMN 和 cALD 星形胶质细胞 该提案是创新的，因为它通过确定 首次发现代谢途径差异调节人类 AMN 和 cALD 的炎症反应 所提出的研究具有重要意义，因为导致不太严重的细胞机制。 即使在四十年后，AMN 或致命的 cALD 表型对相同 ABCD1 突变的反应仍然未知 X-ALD 中基因缺陷的识别影响：随着 X-ALD 后新诊断病例的增加。 2016年被添加到联邦新生儿筛查清单中，迫切需要确定新的目标来开发 对于目前尚无令人满意的治疗方法的 AMN 和 cALD，有有效的治疗方法。