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: 10645207
• 负责人: Jaspreet Singh
• 金额: $ 35.37万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10645207
• 关键词: Accounting; Address; Adrenoleukodystrophy; Adrenomyeloneuropathy; Allogenic; Amino Acids; Area; Astrocytes; Autologous; Automobile Driving; Autopsy; Benign; Biochemical; Body Fluids; Brain; Brain Diseases; CRISPR/Cas technology; Cells; Cerebrum; Citric Acid Cycle; Clinical; Clinical Course of Disease; DNA Sequence Alteration; Data; Defect; Demyelinations; Development; Diagnosis; Disease; Disease Progression; Distant; Drug Design; Energy Metabolism; Exposure to; Female; Fibroblasts; Genes; Genetic Diseases; Genotype; Glucose; Glycolysis; Goals; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Inborn Errors of Metabolism; Inflammatory; Inflammatory Response; Inherited; Investigation; Knock-in; Knowledge; Laboratories; Life; Link; Membrane Proteins; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Mission; Mitochondria; Modeling; Mutation; National Institute of Neurological Disorders and Stroke; Neonatal Screening; Nerve Degeneration; Nervous System; Neurodegenerative Disorders; Neurologic; Neurons; Patients; Phenotype; Plasma; Procedures; Protein Import; Public Health; Recommendation; Regulation; Reporting; Research; Respiration; Risk; Role; Siblings; Source; Specific qualifier value; Sphingolipids; Spinal Cord Diseases; Stimulus; Supplementation; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Translating; Up-Regulation; Very Long Chain Fatty Acid; axonopathy; clinical development; clinical predictors; comparative; cytokine; disease phenotype; drug development; effective therapy; expectation; forging; improved; in vivo; induced pluripotent stem cell; innovation; male; metabolomics; mitochondrial dysfunction; motor disorder; nervous system disorder; new therapeutic target; novel; peroxisome; prognostic indicator; response; screening panel; screening program; stem cell gene therapy; therapeutically effective; transcriptome; white matter; Accounting; Address; Adrenoleukodystrophy; Adrenomyeloneuropathy; Allogenic; Amino Acids; Area; Astrocytes; Autologous; Automobile Driving; Autopsy; Benign; Biochemical; Body Fluids; Brain; Brain Diseases; CRISPR/Cas technology; Cells; Cerebrum; Citric Acid Cycle; Clinical; Clinical Course of Disease; DNA Sequence Alteration; Data; Defect; Demyelinations; Development; Diagnosis; Disease; Disease Progression; Distant; Drug Design; Energy Metabolism; Exposure to; Female; Fibroblasts; Genes; Genetic Diseases; Genotype; Glucose; Glycolysis; Goals; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Inborn Errors of Metabolism; Inflammatory; Inflammatory Response; Inherited; Investigation; Knock-in; Knowledge; Laboratories; Life; Link; Membrane Proteins; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Mission; Mitochondria; Modeling; Mutation; National Institute of Neurological Disorders and Stroke; Neonatal Screening; Nerve Degeneration; Nervous System; Neurodegenerative Disorders; Neurologic; Neurons; Patients; Phenotype; Plasma; Procedures; Protein Import; Public Health; Recommendation; Regulation; Reporting; Research; Respiration; Risk; Role; Siblings; Source; Specific qualifier value; Sphingolipids; Spinal Cord Diseases; Stimulus; Supplementation; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Translating; Up-Regulation; Very Long Chain Fatty Acid; axonopathy; clinical development; clinical predictors; comparative; cytokine; disease phenotype; drug development; effective therapy; expectation; forging; improved; in vivo; induced pluripotent stem cell; innovation; male; metabolomics; mitochondrial dysfunction; motor disorder; nervous system disorder; new therapeutic target; novel; peroxisome; prognostic indicator; response; screening panel; screening program; stem cell gene therapy; therapeutically effective; transcriptome; white matter

PROJECT SUMMARY/ABSTRACT The mechanism of disease progression from benign to fatal phenotypes in X-linked adrenoleukodystro- phy remains unknown and there is no satisfactory cure for the disease. 60% of male X-ALD patients develop fatal cerebral disease (cALD) while the remaining 40% develop milder adrenomyeloneuropathy (AMN) charac- terized by axonopathy. The primary genetic defect in X-ALD (mutation/deletion in ABCD1 gene) and the bio- chemical defect (accumulation of very long chain fatty acid; C>22:0 in plasma and tissues) cannot predict the onset of AMN or cALD. The long-term goal is to contribute to the development of novel clinically useful, mecha- nism-based prognostic indicators and therapeutic options for X-ALD. The overall objective for this application is to determine differential metabolic energy metabolism underlying phenotype variability (AMN vs cALD) in the human astrocytes of male X-ALD phenotypes. The central hypothesis is that altered metabolic reprogramming underlies the differential phenotype development in AMN and cALD astrocytes. These astrocytes were differen- tiated from induced pluripotent stem cells (iPSCs), which, in turn, were generated by reprogramming of human control, AMN and cALD patient-derived fibroblasts. This hypothesis is supported by untargeted metabolomics pilot data identifying metabolites altered between healthy-control and cALD phenotype postmortem brain and between AMN and cALD astrocytes. Within the cALD brain white matter, unique metabolite changes were rec- orded between distant normal looking areas and areas adjacent to the plaque suggesting an association with disease progression. OXPHOS and glycolysis were found to be decreased (low metabolic state) in human cALD astrocytes. This low metabolic state suggests a role for novel alternative source(s) of fuel driving the progression to cALD phenotype in astrocytes. The rationale for the proposed research is that a mechanistic modelling of aberrant energy metabolism in AMN and cALD astrocytes will provide a basis for predicting disease progression and new opportunities for identification of targets for novel therapeutic drug design. The central hypothesis will be tested by pursuing two specific aims: 1) Determine the role of metabolic reprogramming in newly “forged” AMN and cALD astrocytes; and 2) Determine the contribution of mitochondrial dysfunction in metabolic repro- gramming in AMN and cALD astrocytes. The approach will take advantage of control, AMN and cALD astrocytes recently generated from iPSCs in the laboratory. This proposal is innovative because it departs from the status quo by identifying for the first time, metabolic pathways differentially altered 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 identifica- tion of gene defect in X-ALD. Successful completion of the proposed research is expected to provide a necessary conceptual framework for the subsequent development of clinically effective strategies for predicting disease progression and improving current limited treatment options.

项目摘要/摘要 X连锁性肾上腺素的疾病进展的机制从良性到致命表型的发展 PHY仍然未知，没有满意厂治疗该疾病。 60％的男性X-ALD患者发展 致命的大脑疾病（CALD），而其余40％的米勒肾上腺病（AMN）特征 由轴突病变。 X-ald（ABCD1基因中的突变/缺失）和Bio-的主要遗传缺陷 化学缺陷（在血浆和组织中的长链脂肪酸的积累； C> 22：0）无法预测 AMN或CALD的发作。长期的目标是为新型临床有用的机甲发展​​做出贡献 X-Ald的基于Nism的预后指标和治疗选择。该应用程序的总体目标是 确定差异代谢能量代谢基础表型的可变性（AMN与CALD） 男性X-Ald表型的人类星形胶质细胞。中心假设是改变了代谢重编程 基础是AMN和CALD星形胶质细胞中的差异表型发育。这些星形胶质细胞不同 由诱导的多能干细胞（IPSC）产生的，而这些干细胞是通过重新编程而产生的 对照，AMN和CALD患者衍生的成纤维细胞。该假设得到了不靶向的代谢组学的支持 识别健康控制和CALD表型事后大脑和CALD表型之间发生变化的代谢物的试验数据和 在AMN和CALD星形胶质细胞之间。在CALD脑白质中，独特的代谢物变化是恢复的 在遥远的正常面积和与斑块附近的区域之间进行命令，这表明与 疾病进展。在人CALD中发现了Oxphos和Glycolysis改善（低代谢状态） 星形胶质细胞。这种低代谢状态表明了新型替代来源的燃料驱动进展的作用 在星形胶质细胞中表型。拟议研究的理由是 AMN和CALD星形胶质细胞中的异常能量代谢将为预测疾病进展提供基础 以及确定新型热药设计目标的新机会。中心假设将 通过追求两个具体目标来测试：1）确定代谢重编程在新“伪造”中的作用 AMN和CALD星形胶质细胞； 2）确定线粒体功能障碍在代谢再生中的贡献 在AMN和CALD星形胶质细胞中进行语法。该方法将利用控制，AMN和CALD星形胶质细胞 最近由IPSC在实验室中产生。该提议具有创新性，因为它偏离了身份 通过首次识别人AMN和CALD星形胶质细胞中的代谢途径不同，代谢途径有所不同。 拟议的研究很重要，因为导致AMN或致命不太严重的细胞机制 CALD表型响应相同的ABCD1突变，即使在鉴定后四十年 - 基因缺陷的X-ald。预计拟议研究的成功完成将提供必要的 随后开发临床有效策略来预测疾病的概念框架 进展并改善当前有限的治疗选择。