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.

项目总结/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）疾病仍然未知。60%的男性X-ALD患者发生致命的脑 神经炎症（cALD），而其余发展为轻度肾上腺脊髓神经病（AMN），其特征在于 无神经炎症的轴突病。X-ALD的主要遗传缺陷（ABCD 1基因突变/缺失） 和生化缺陷（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的发作。我们的长期目标是剖析 X-ALD中的差异表型发展。本申请的目的是确定代谢途径 这是AMN和cALD人星形胶质细胞中不同神经炎症反应的基础。这些星形胶质细胞 从诱导多能干细胞（iPSC）分化而来，而iPSC又是通过重编程产生的。 人对照、AMN和cALD患者来源的成纤维细胞。代谢重编程正在成为一种新的 炎症反应的调节剂。星形胶质细胞依靠线粒体呼吸（OXPHOS）进行代谢。 需要，但在神经炎症环境下切换到糖酵解，以促进生物合成途径， 炎症介质。我们的初步概念验证数据，与非靶向代谢组学，确定 代谢物在健康对照和死后脑的cALD表型之间改变。在cALD大脑中是白色 物质，独特的代谢物的变化之间记录遥远的正常外观的地区和地区相邻 斑块表明与疾病进展有关。我们发现OXPHOS和糖酵解 降低（低代谢状态）的人cALD星形胶质细胞，尽管较高的炎症反应。这种低 代谢状态提示了新的替代燃料来源在驱动神经炎症反应中的作用。 cALD星形胶质细胞。我们的中心假设是cALD星形胶质细胞中的代谢重编程驱动了它们的生长。 在cALD中，促炎性转变是神经炎性疾病进展的基础。为了验证我们的假设 我们提出了两个具体的目标：1）阐明负责炎症反应的代谢重编程， cALD星形胶质细胞的反应。2)为了确定功能障碍的线粒体是否在炎症性质中发挥作用， cALD星形胶质细胞我们将利用对照、AMN和cALD星形胶质细胞从iPSC中产生， 这些研究的实验室。这项建议是创新的，因为它通过确定 第一次，代谢途径差异调节人类AMN和cALD的炎症反应 星形胶质细胞。拟议的研究是重要的，因为导致不太严重的细胞机制 即使在40年后，对相同ABCD 1突变的AMN或致死性cALD表型的反应仍然未知。 X-ALD基因缺陷的鉴定。影响：随着X-ALD后新诊断病例率的上升 在2016年被添加到联邦新生儿筛查名单中，迫切需要确定新的目标来开发 有效的治疗AMN和cALD的方法，对于这些方法还没有令人满意的治疗方法。