Effects of ABCD1 Deficiency on Endothelial Function and Permeability to Leukocytes in Cerebral X-linked Adrenoleukodystrophy

ABCD1 缺陷对脑 X 连锁肾上腺脑白质营养不良患者内皮功能和白细胞通透性的影响

• 批准号: 10657587
• 负责人: Patricia Leonor Musolino
• 金额: $ 39.48万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657587
• 关键词: Acute; Address; Adhesions; Adrenoleukodystrophy; Affect; Age of Onset; Animal Model; Astrocytes; Binding; Biological; Biological Assay; Biological Models; Blood - brain barrier anatomy; Blood brain barrier dysfunction; Brain; Cell Adhesion Molecules; Cell model; Cells; Cerebrum; Cessation of life; Coculture Techniques; Code; Data; Defect; Demyelinations; Development; Disease; Disease Progression; Disease model; Dose; Down-Regulation; Electrical Resistance; Endothelium; Event; Extravasation; Female; Functional disorder; Future; Gene Dosage; Gene Mutation; Gene Silencing; Gene Targeting; Genes; Genetic Transcription; Genotype; Heterozygote; Human; Individual; Infiltration; Inflammatory; Inherited; Knowledge; Laboratories; Leukocytes; Maintenance; Mediating; Modeling; Molecular; Mutation; Oncogenes; Pathway interactions; Patients; Penetrance; Pericytes; Permeability; Phenotype; Preventive; Preventive treatment; Proteins; Regulation; Regulatory Pathway; Role; Signal Pathway; Stress; TGFB1 gene; Techniques; Testing; Tight Junctions; Toxic effect; Transcriptional Regulation; Transforming Growth Factor beta; Vascular Endothelium; Vegetative States; Very Long Chain Fatty Acid; Viral; Work; blood-brain barrier disruption; blood-brain barrier function; blood-brain barrier permeabilization; brain dysfunction; brain endothelial cell; cerebral degeneration; cytokine; differential expression; early childhood; experimental study; imaging study; induced pluripotent stem cell; insight; male; migration; mortality; nervous system disorder; neuroinflammation; novel; novel therapeutic intervention; peroxisome; prevent; promoter; small molecule; targeted treatment; tool; transcription factor; transcriptome sequencing; validation studies

X-linked adrenoleukodystrophy (ALD) is a devastating neurologic disorder caused by mutations in the ABCD1 gene characterized by the accumulation of very long-chain fatty acids that affects 1:17,000 individuals in the U.S. Approximately 60% of male patients with ALD will convert to a devastating rapidly progressive form of inflammatory demyelination that leads to incapacitation or death within 2-3 years (cerebral ALD). Age of onset and phenotype varies even among individuals with the same mutation but a constant initial event in cerebral ALD is blood brain barrier (BBB) disruption with migration of leukocytes to the brain. The precise molecular and cellular mechanisms controlling BBB function during the course of ALD disease progress remain poorly understood given lack of cellular or animal models that faithfully recapitulate cerebral ALD. To address this critical knowledge gap, Dr. Musolino’s laboratory recently developed an ALD ex-vivo model system using gene-editing strategies and human brain microvascular endothelial cells. Dr. Musolino’s initial studies provide evidence that loss of ABCD1 directly impairs brain endothelial barrier integrity by increasing TGFβ1 levels in a manner correlated with severe transcriptional downregulation of Claudin 5 and increased permeability to small molecules. These alterations precede the accumulation of very-long chain fatty acids suggesting that the BBB dysfunction is not a direct consequence of their accumulation. As with patients, in addition to ABCD1 deficiency in this model, a second event, endothelial activation by inflammatory cytokines or flow sheer stress, is necessary to increase the permeability to leukocytes. Building upon these strong preliminary data Dr. Musolino’s hypothesizes that levels of ABCD1 expression in brain microvascular endothelium controls transcriptional regulation of tight junction proteins via TGFβ1-regulated pathways and determines the permeability to leukocytes during endothelial activation in a dose-dependent manner. To test this hypothesis Dr. Musolino will probe the effect of ABCD1 deficiency upon the BBB by (1) Identifying the molecular mechanisms governing tight junction disruption and increased permeability of ABCD1- deficient brain endothelium (Aim 1), (2) Determining functional consequences of downregulation of tight junction proteins and main regulatory pathways (Aim 2), and (3) Quantifying ABCD1 gene-dose effect on endothelial barrier function (Aim 3). Upon successful completion of these studies Dr. Musolino will have leverage the ability to model the impact of a single-gene mutation to unravel the mechanisms governing the traffic of cells across the BBB in ALD setting forth a strategy to identify the molecular and cell biological mechanisms underlying the conversion to cerebral disease, develop functional assays to test novel therapeutic approaches, and inform the field of neuroinflammation.

X连锁的肾上腺素肌营养不良（ALD）是由突变引起的毁灭性神经系统疾病 ABCD1基因的特征是影响1：17,000个个体的长链脂肪酸的积累 在美国，大约60％的ALD患者转化为毁灭性的迅速编程的男性患者 在2-3内（大脑ALD）内形成脱后脱髓鞘或死亡 发作和表型的偶数甚至在具有相同突变的个体之间有所不同，但在 小脑ALD是血液屏障（BBB）的破坏，白细胞向大脑迁移 在ALD疾病进步过程中控制BBB功能的分子和细胞机制 鉴于缺乏忠实地缩写ALD的细胞或动物模型，因此仍然很糟糕。 为了解决这个关键的知识差距，Musolino博士的实验室最近开发了一个ALD Ex-Vivo模型 使用基因编辑策略和人脑微血管内皮细胞的系统 研究提供了证据表明，ABCD1的丧失直接通过增加而直接损害脑室内治疗屏障完整性 tgfels的方式与claudred的转录下调相关 对小分子的渗透性。 与BBB功能障碍一样，与患者的积累直接奉献在一起。 在该模型中增加了ABCD1缺乏症，第二个事件，炎症细胞因子的内皮激活 或流动的压力是增加对牢固的渗透性的必要条件。 初步数据Musolino博士的假设是脑微血管内皮中ABCD1表达的水平 通过TGFβ1调控途径来控制紧密连接蛋白的转录调控，并确定您 在剂量依赖性方式中对白细胞激活期间白细胞的渗透性。 为了检验该假设 管理紧密连接学和ABCD1-渗透性提高的分子机制 缺乏大脑内皮（AIM 1），（2）确定紧密下调的功能结构。 封闭蛋白和主要规律性途径（AIM 2）和（3）量化ABCD1基因剂量对 内皮屏障功能（AIM 3）。 利用建模单基因突变的影响的能力，以揭示控制您的机制 细胞在ALD中跨BBB的流量制定了识别分子和细胞生物学的策略 趋于趋于脑疾病的机制，开发功能性测定以测试新型治疗疗法 接近并告知神经炎症领域。

项目成果

Neurofilament light chain as a potential biomarker for monitoring neurodegeneration in X-linked adrenoleukodystrophy.

• DOI: 10.1038/s41467-021-22114-2
• 发表时间: 2021-03-22
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Weinhofer I;Rommer P;Zierfuss B;Altmann P;Foiani M;Heslegrave A;Zetterberg H;Gleiss A;Musolino PL;Gong Y;Forss-Petter S;Berger T;Eichler F;Aubourg P;Köhler W;Berger J
• 通讯作者: Berger J

Hematopoietic stem-cell gene therapy is associated with restored white matter microvascular function in cerebral adrenoleukodystrophy.

• DOI: 10.1038/s41467-023-37262-w
• 发表时间: 2023-04-05
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Lauer, Arne;Speroni, Samantha L.;Choi, Myoung;Da, Xiao;Duncan, Christine;McCarthy, Siobhan;Krishnan, Vijai;Lusk, Cole A.;Rohde, David;Hansen, Mikkel Bo;Kalpathy-Cramer, Jayashree;Loes, Daniel J.;Caruso, Paul A.;Williams, David A.;Mouridsen, Kim;Emblem, Kyrre E.;Eichler, Florian S.;Musolino, Patricia L.
• 通讯作者: Musolino, Patricia L.

The Genetic Landscape of Ischemic Stroke in Children - Current Knowledge and Future Perspectives.

• DOI: 10.1016/j.spen.2022.100999
• 发表时间: 2022-09
• 期刊: Seminars in pediatric neurology
• 影响因子: 2.7
• 作者: M. Hausman-Kedem;Rachelle Herring;M. D. Torres;Jonathan D. Santoro;M. Kaseka;C. Vargas;Giulia Amico-Giulia-Am