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，神经胶质细胞激活和突触丢失。有复发和永久性残疾。尽管 针对免疫系统中的细胞的治疗的成功，对针对细胞的治疗的需求尚未得到满足 在中枢神经系统(CNS)修复残疾。四个观察为一种新的方法提供了理由 多发性硬化症患者的神经变性：1)多发性硬化症患者的残疾是异质性的，并且明显的残疾(行走， 视觉、认知、协调)由不同的中枢神经系统区域提供服务，2)即使在健康的大脑中，给定的中枢神经系统细胞 大脑不同区域的基因表达类型不同，3)女性和男性会影响残疾 病情恶化，4)衰老与残疾进展相一致。在这里，我们将使用特定于细胞、特定于区域和 寻找针对多发性硬化症女性和男性不同残疾的最佳治疗目标的具体性别方法。 多发性硬化症患者从床到床：性别差异的临床观察 临床前水平然后作为针对不同性别设计的试验重新转化到临床。女性和女性的临床前使用 实验性自身免疫性脑脊髓炎(EAE)雄性小鼠体内区域特异性磁共振成像 萎缩，每个区域的神经病理，每个区域不同的中枢神经系统细胞的RNA测序， 细胞特异性高差异表达通路中TOP基因的免疫组织化学验证 目的基因的条件性敲除(CKO)逆转表型，目的基因的敲除 逆转表型的药物治疗。遗传(CKO与WT)和/或药物的影响 (治疗与安慰剂)逆转基因表达的干预措施在不同性别之间确定。人类的MS数据 指导三个检查点的临床前研究：i)女性和男性多发性硬化症患者的MRI显示性别差异 在亚结构萎缩中，优先考虑有萎缩的EAE区域，ii)单核RNA序列分析在女性和 男性MS患者在EAE中优先选择感兴趣的基因途径，III)免疫组织化学 在女性和男性中，使用MS尸检组织验证EAE的免疫组织化学。取代使用 雌性小鼠与雄性小鼠(如上所述)使用性腺切除的小鼠与性腺完整的小鼠的对比将揭示 性激素的激活作用。使用四种核心基因型鼠将揭示性染色体效应与 发育荷尔蒙效应。使用幼鼠和老年鼠将揭示衰老的影响。 这项R35提案将：1)扩展我们星形胶质细胞的细胞特异性和区域特异性转录 和少突胶质细胞到小胶质细胞和神经元，与细胞：小鼠的细胞相互作用被双标记到 显示同一小鼠同一区域内两种不同细胞类型的基因表达变化，以及2)确定 是否存在性别和/或年龄对差异表达最多的特定细胞和特定区域的影响 小路。总而言之，R35提案将我们的研究带到了一个新的水平：通过年龄互动来识别性别 在细胞特异性和区域特异性转录，神经病理学和核磁共振亚基结构萎缩。