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.

项目摘要/摘要 X连锁肾上腺脑白质营养不良从良性表型向致命性表型转化的机制 PHY仍然未知，也没有令人满意的治疗方法。60%的男性X-ALD患者发生 致命的脑部疾病(CALD)，而其余40%的人发展为较轻的肾上腺脊髓神经病(AMN)特征- 以轴索病为特征。X-ALD(ABCD1基因突变/缺失)的原发遗传缺陷及其生物学意义 化学缺陷(血浆和组织中超长链脂肪酸的积累；C&gt；22：0)不能预测 AMN或CALD的发病。长期目标是为临床上有用的新型机械设备的开发做出贡献。 基于NISM的X-ALD预后指标和治疗选择。这个应用程序的总体目标是 确定表型变异(AMN与CALD)背后的不同代谢能量代谢 男性X-ALD表型的人星形胶质细胞。中心假说是改变新陈代谢重新编程 这是AMN和Cald星形胶质细胞不同表型发育的基础。这些星形细胞是不同的- 来自诱导多能干细胞(IPSCs)，而IPSCs又是通过重新编程人类产生的 对照组、AMN和CALD患者来源的成纤维细胞。这一假说得到了非靶向代谢组学的支持 鉴定在健康对照和CALD表型之间改变的代谢物的先导数据死后脑和 在AMN和Cald星形胶质细胞之间。在Cald脑白质内，记录了独特的代谢物变化。 在看起来正常的远处区域和邻近斑块的区域之间排列的，表明与 疾病的发展。在人类CALD中发现氧磷酸盐和糖酵解减少(低代谢状态) 星形胶质细胞。这种低代谢状态表明了推动进展的新的替代燃料来源(S)的作用 星形胶质细胞的Cald表型。这项拟议研究的基本原理是， AMN和Cald星形胶质细胞能量代谢异常将为预测疾病进展提供基础 以及为新的治疗药物设计确定靶点的新机会。中心假说将 通过追求两个具体目标来测试：1)确定新陈代谢重新编程在新的“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再生中的作用。 在AMN和Cald星形胶质细胞中摄取。该方法将利用Control、AMN和Cald星形胶质细胞 最近从实验室的ipscs中产生的。这项提议是创新的，因为它脱离了现状 通过首次鉴定，人类AMN和Cald星形胶质细胞的代谢途径发生了不同的变化。 提出的研究具有重要意义，因为导致AMN较轻或致命性较低的细胞机制(S) 对相同ABCD1突变的CALD表型即使在鉴定40年后仍未知- X-ALD中基因缺陷的检测。成功完成拟议的研究预计将提供必要的 随后开发临床有效的疾病预测策略的概念框架 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