Neurodegeneration Underlying Distinct Disabilities in Multiple Sclerosis Using a Cell-Specific, Region-Specific, and Sex-Specific Approach

使用细胞特异性、区域特异性和性别特异性方法研究多发性硬化症中明显残疾的神经退行性变

• 批准号: 10663020
• 负责人: RHONDA R VOSKUHL
• 金额: $ 88.15万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2031-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10663020
• 关键词: Age; Aging; Astrocytes; Atrophic; Autoimmune; Autopsy; Back; Brain; Brain region; Cell Communication; Cells; Central Nervous System; Clinic; Clinical; Cognition; Data; Demyelinations; Development; Experimental Autoimmune Encephalomyelitis; Female; Four Core Genotypes; Gene Expression; Genes; Genetic; Gonadal Steroid Hormones; Hormones; Human; Immune system; Immunohistochemistry; Inflammation; Intervention; Label; Magnetic Resonance Imaging; Microglia; Modeling; Multiple Sclerosis; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Oligodendroglia; Pathway interactions; Pharmacological Treatment; Phenotype; Placebos; Process; Relapsing-Remitting Multiple Sclerosis; Research; Secondary Progressive Multiple Sclerosis; Sex Chromosomes; Sex Differences; Synapses; Tissues; Translating; Validation; Vision; Walking; Woman; age effect; axon injury; bench to bedside; cell type; conditional knockout; differential expression; disability; disability impact; glial activation; in vivo; interest; knock-down; male; men; multiple sclerosis patient; neuropathology; neuroprotection; novel; novel strategies; optimal treatments; pharmacologic; pre-clinical; pre-clinical research; repaired; sex; single nucleus RNA-sequencing; success; transcriptome sequencing; transcriptomics; trial design; young adult

Multiple sclerosis (MS) is an autoimmune, neurodegenerative disease with inflammation, demyelination, axonal damage, glial activation and synaptic loss. There are relapses and permanent disabilities. Despite success of treatments targeting cells in the immune system, there is an unmet need for treatments targeting cells in central nervous system (CNS) to repair disabilities. Four observations provide rationale for a new approach to neurodegeneration in MS: 1) MS patients are heterogenous in their disabilities, and distinct disabilities (walking, vision, cognition, coordination) are served by different CNS regions, 2) Even in healthy brain, a given CNS cell type differs in gene expression from one brain region to another, 3) Being female versus male impacts disability worsening, and 4) Aging aligns with disability progression. Here, we will use a cell-specific, region-specific, and sex-specific approach to discover optimal treatment targets for distinct disabilities in MS women and men. Bedside to Bench to Bedside in MS: Clinical observations of sex differences are investigated at the preclinical level then translated back to the clinic as trials designed for each sex. Preclinical use of female and male mice with experimental autoimmune encephalomyelitis (EAE) entails in vivo MRI for region-specific atrophy, neuropathology of each region, RNA-sequencing of distinct CNS cells in each region, immunohistochemistry validation of top genes in highly differentially expressed pathways, cell-specific conditional knockout (CKO) of target genes to reverse phenotype, and knockdown of target genes with pharmacologic treatment to reverse phenotype. The effect of genetic (CKO vs WT) and/or pharmacologic (treatment vs placebo) interventions on reversal of gene expression is determined in each sex. Human MS data guide preclinical research at three checkpoints: i) MRI in females and males with MS revealing sex differences in substructure atrophy prioritize regions in EAE with atrophy, ii) Single nuclei RNA-seq analyses in females and males with MS revealing gene pathways of interest prioritize gene pathways in EAE, iii) immunohistochemistry in females and males using MS postmortem tissues validate immunohistochemistry in EAE. Substitution of use of female versus male mice (as above) with use of gonadectomized versus gonadally intact mice will reveal activational effects of sex hormones. Use of Four Core Genotype mice will reveal sex chromosome effects versus developmental hormone effects. Use of young versus old mice will reveal the effect of aging. This R35 proposal will: 1) Extend our cell-specific and region-specific transcriptomics in astrocytes and oligodendrocytes to microglia and neurons, with cell:cell interactions revealed in mice double-labelled to show gene expression changes in two distinct cell types in the same region in the same mouse, and 2) Determine if there are effects of sex and/or age on the most differentially expressed cell-specific and region-specific pathways. In summary, this R35 proposal takes our research to the next level: Identifying sex by age interactions in cell-specific and region-specific transcriptomics, neuropathology, and substructure atrophy on MRI.

多发性硬化症 (MS) 是一种自身免疫性神经退行性疾病，伴有炎症、脱髓鞘、 轴突损伤、神经胶质激活和突触损失。有复发和永久性残疾。尽管 针对免疫系统中的细胞的治疗取得了成功，但针对细胞的治疗的需求尚未得到满足 中枢神经系统（CNS）修复残疾。四项观察结果为新方法提供了理由 MS 中的神经退行性变：1) MS 患者的残疾情况各不相同，且残疾程度不同（行走、 视觉、认知、协调）由不同的 CNS 区域提供服务，2）即使在健康的大脑中，给定的 CNS 细胞 不同大脑区域的基因表达类型不同，3) 女性与男性会影响残疾 恶化，以及 4) 衰老与残疾进展一致。在这里，我们将使用特定于细胞、特定于区域和 针对多发性硬化症女性和男性不同残疾的最佳治疗目标。 MS 的床边到工作台到床边：性别差异的临床观察在 然后，临床前水平转化回临床，作为针对每种性别设计的试验。临床前使用女性和 患有实验性自身免疫性脑脊髓炎 (EAE) 的雄性小鼠需要进行体内 MRI 检测区域特异性 萎缩、每个区域的神经病理学、每个区域不同 CNS 细胞的 RNA 测序、 对高度差异表达途径、细胞特异性的顶级基因进行免疫组织化学验证 目标基因的条件性敲除（CKO）以逆转表型，以及用 药物治疗以逆转表型。遗传（CKO 与 WT）和/或药理学的影响 （治疗与安慰剂）对基因表达逆转的干预措施是在每个性别中确定的。人类 MS 数据 在三个检查点指导临床前研究：i) 女性和男性 MS 患者的 MRI 显示性别差异 在亚结构萎缩中，优先考虑 EAE 中萎缩的区域，ii) 女性中的单核 RNA-seq 分析和 患有 MS 的男性揭示了感兴趣的基因通路优先考虑 EAE 中的基因通路，iii) 免疫组织化学 使用 MS 死后组织在女性和男性中验证 EAE 中的免疫组织化学。替代使用 使用去性腺切除小鼠与性腺完整小鼠的雌性小鼠与雄性小鼠（如上所述）的比较将揭示 性激素的激活作用。使用四核心基因型小鼠将揭示性染色体效应 发育激素的影响。使用年轻小鼠与年老小鼠将揭示衰老的影响。 该 R35 提案将： 1) 扩展我们在星形胶质细胞中的细胞特异性和区域特异性转录组学 和少突胶质细胞到小胶质细胞和神经元，在双标记小鼠中揭示了细胞与细胞之间的相互作用 显示同一小鼠同一区域中两种不同细胞类型的基因表达变化，以及 2) 确定 性别和/或年龄是否对差异最大的细胞特异性和区域特异性表达有影响 途径。总之，这个 R35 提案将我们的研究提升到了一个新的水平：通过年龄相互作用识别性别 细胞特异性和区域特异性转录组学、神经病理学和 MRI 亚结构萎缩。