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、神经胶质活化和突触丢失。有复发和永久性残疾。尽管 尽管靶向免疫系统中的细胞的治疗取得了成功，但对靶向细胞的治疗存在未满足的需求 在中枢神经系统（CNS）修复残疾。四个观察结果为采取新的方法提供了依据， MS中的神经变性：1）MS患者的残疾是异质性的，并且不同的残疾（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 类型不同的基因表达从一个大脑区域到另一个，3）女性与男性影响残疾 恶化，以及4）衰老与残疾进展一致。在这里，我们将使用细胞特异性、区域特异性和 性别特异性方法，以发现MS女性和男性不同残疾的最佳治疗目标。 MS中的床边到长凳到床边：在研究中研究了性别差异的临床观察结果。 临床前水平，然后翻译回临床试验设计为每个性别。临床前使用女性和 患有实验性自身免疫性脑脊髓炎（EAE）的雄性小鼠需要在体内MRI进行区域特异性 萎缩，每个区域的神经病理学，每个区域中不同CNS细胞的RNA测序， 高度差异表达途径中顶级基因的免疫组织化学验证，细胞特异性 靶基因的条件性敲除（CKO）以逆转表型，以及用 逆转表型的药物治疗。遗传（CKO vs WT）和/或药理学的影响 在每种性别中确定对逆转基因表达的干预（治疗与安慰剂）。人MS数据 在三个检查点指导临床前研究：i）患有MS的女性和男性的MRI显示性别差异 在亚结构萎缩中，优先考虑EAE中具有萎缩区域，ii）雌性中的单核RNA-seq分析， 显示感兴趣的基因途径的MS男性优先考虑EAE中的基因途径，iii）免疫组织化学 在雌性和雄性中使用MS死后组织验证EAE中的免疫组织化学。替代使用 使用性腺切除小鼠与性腺完整小鼠的雌性与雄性小鼠（如上所述）的比较将揭示 性激素的激活作用。使用四个核心基因型小鼠将揭示性染色体效应， 发育激素的影响。使用年轻小鼠与老年小鼠将揭示衰老的影响。 这项R35提案将：1）在星形胶质细胞中扩展我们的细胞特异性和区域特异性转录组学 和少突胶质细胞的小胶质细胞和神经元，与细胞：细胞的相互作用，揭示了在小鼠双标记， 显示同一小鼠中相同区域中两种不同细胞类型的基因表达变化，以及2）确定 如果性别和/或年龄对最差异表达的细胞特异性和区域特异性 途径。总之，R35提案将我们的研究带到了一个新的水平：通过年龄相互作用识别性别 在细胞特异性和区域特异性转录组学、神经病理学和MRI上的亚结构萎缩中。