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

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.

项目概要/摘要 X连锁肾上腺脑白质营养不良疾病从良性表型进展到致命表型的机制 phy 仍然未知，并且对该疾病没有令人满意的治疗方法。 60% 的男性 X-ALD 患者患有 X-ALD 致命性脑病 (cALD)，而其余 40% 则出现较轻的肾上腺脊髓神经病 (AMN) 特征 因轴突病而死亡。 X-ALD 的主要遗传缺陷（ABCD1 基因突变/缺失）和生物- 化学缺陷（极长链脂肪酸的积累；血浆和组织中 C>22:0）无法预测 AMN 或 cALD 发作。长期目标是为开发新型临床有用的机械设备做出贡献 基于 Nism 的 X-ALD 预后指标和治疗选择。该应用程序的总体目标是 确定表型变异（AMN 与 cALD）背后的差异代谢能量代谢 男性 X-ALD 表型的人类星形胶质细胞。中心假设是改变代谢重编程 是 AMN 和 cALD 星形胶质细胞差异表型发育的基础。这些星形胶质细胞是不同的 源自诱导多能干细胞 (iPSC)，而 iPSC 又是通过对人类细胞进行重新编程而产生的 对照、AMN 和 cALD 患者来源的成纤维细胞。这一假设得到非靶向代谢组学的支持 试验数据确定了健康对照和 CALD 表型死后大脑之间代谢物的变化 AMN 和 cALD 星形胶质细胞之间。在 cALD 大脑白质内，发现了独特的代谢变化 在遥远的正常区域和邻近斑块的区域之间排序，表明与 疾病进展。发现人类 cALD 中的 OXPHOS 和糖酵解减少（低代谢状态） 星形胶质细胞。这种低代谢状态表明新的替代燃料来源在推动进展中发挥着作用 星形胶质细胞中的 cALD 表型。所提出的研究的基本原理是，机械模型 AMN 和 cALD 星形胶质细胞异常的能量代谢将为预测疾病进展提供基础 以及识别新型治疗药物设计靶点的新机会。中心假设将 通过追求两个具体目标进行测试：1）确定代谢重编程在新“锻造”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生殖中的贡献 AMN 和 cALD 星形胶质细胞的编程。该方法将利用对照、AMN 和 cALD 星形胶质细胞 最近在实验室中由 iPSC 生成。这个提议是创新的，因为它脱离了现状 通过首次确定人类 AMN 和 cALD 星形胶质细胞中代谢途径的差异改变。 拟议的研究意义重大，因为导致不太严重的 AMN 或致命的细胞机制 即使在鉴定后四十年，对相同 ABCD1 突变的 cALD 表型仍然未知。 X-ALD 中基因缺陷的灰化。成功完成拟议的研究预计将为 后续开发临床有效预测疾病策略的概念框架 进展和改善目前有限的治疗选择。

项目成果

MicroRNA and metabolomics signatures for adrenomyeloneuropathy disease severity.

• DOI: 10.1002/jmd2.12323
• 发表时间: 2022-11
• 期刊: JIMD reports
• 作者: Turk, Bela Rui;Poisson, Laila Marie;Nemeth, Christina Linnea;Goodman, Jordan;Moser, Ann B;Jones, Richard Owen;Fatemi, Ali;Singh, Jaspreet
• 通讯作者: Singh, Jaspreet

IPSC-Derived Astrocytes to Model Neuroinflammatory and Metabolic Responses in X-linked Adrenoleukodystrophy.

IPSC 衍生的星形胶质细胞可模拟 X 连锁肾上腺脑白质营养不良的神经炎症和代谢反应。

• DOI: 10.26502/jbb.2642-91280091
• 发表时间: 2023
• 期刊: Journal of biotechnology and biomedicine
• 作者: Parasar,Parveen;Kaur,Navtej;Singh,Jaspreet
• 通讯作者: Singh,Jaspreet