"Lnc"ing XIST Ribonucleoprotein Particles to Female Sex-Attributed Biases in Autoimmunity

“Lnc”ing XIST 核糖核蛋白颗粒对自身免疫中女性性别归因的偏见

• 批准号: 10707968
• 负责人: Diana Remy Dou
• 金额: $ 10.56万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-21 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10707968
• 关键词: Address; Advisory Committees; Affect; American; Antigen-Antibody Complex; Antigens; Atlases; Authorization documentation; Autoantibodies; Autoantigens; Autoimmune; Autoimmune Diseases; Autoimmunity; Blood Tests; CRISPR interference; CRISPR screen; Cells; Chemicals; Chromosome 11; Chromosomes; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complement; Complex; Custom; Development; Developmental Biology; Diagnostic; Disease; Disease model; Dosage Compensation (Genetics); Environment; Epigenetic Process; Evaluation; Female; Foundations; Functional disorder; Gene Dosage; Genes; Genetic; Genomics; Goals; Heart Diseases; High Prevalence; Hormonal; Hormones; Human; Immune; Immune System Diseases; Immune response; Immune signaling; Immunity; Immunologic Stimulation; In Vitro; Individual; Investigation; Klinefelter' s Syndrome; Link; Malignant Neoplasms; Mentors; Mentorship; Modeling; Monitor; Multiomic Data; Mus; Organism; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Phenotypic Sex; Pilot Projects; Predisposition; Prevalence; Pristane; Production; Proteins; Proteomics; Public Health; Research; Research Personnel; Resolution; Resources; Rheumatoid Arthritis; Ribonucleoproteins; Risk; Role; Scientific Inquiry; Serum; Sex Differences; Societies; Spleen; Splenocyte; Supervision; Systemic Lupus Erythematosus; Systemic Scleroderma; T-Lymphocyte; Techniques; Technology; Testing; Tissues; Training; Transgenic Mice; Transgenic Organisms; Universities; Untranslated RNA; Women' s Health; Work; X Chromosome; authority; autoreactivity; biological sex; candidate validation; career; cell type; cost; design; effective therapy; epigenome; experimental study; human disease; immune activation; in vivo; innovation; male; mouse model; multidisciplinary; multiple omics; novel; particle; potential biomarker; preservation; protein complex; rheumatologist; screening; sex; single cell sequencing; societal costs; therapeutic target; therapeutically effective

Project Summary / Abstract Autoimmune diseases (AD) are the third most prevalent disease in America, disproportionately impacting females 4x more than males. Despite the high prevalence and cost to society, there are few effective treatments and no definitive cure because the genetic factors and environmental triggers for autoimmunity remain poorly defined. Existing studies implicating hormonal differences to explain the sex differences in autoimmunity and immune response fail to reconcile the increased susceptibility to autoimmune diseases in Kleinfelter males, who have two X chromosomes (XXY) like females (XX) and unlike most males (XY). In XX individuals, the long noncoding RNA (lncRNA), XIST (Xist in mice), is required for silencing one of the X’s to achieve gene dosage compensation. This project aims to elucidate the XX-linked preponderance for autoimmune disease development through studying the female-specific lncRNA, XIST, and the proteins associating with XIST in the XIST ribonucleoprotein complex (RNP) as a potential immune complex trigger for autoimmunity. In this proposal, Dr. Dou proposes to test the novel hypothesis that Xist RNPs increase autoimmune risk using three multi-level aims: (1) In vitro, through controlled stimulation of immune cells with Xist RNPs, (2) In vivo, through autoimmune disease modeling in a transgenic Xist mouse wherein male mice viably express Xist RNPs, (3) diagnostically, using autoimmune disease patient serum to test reactivity against XIST RNP proteins. Completion of the project will build a comprehensive model of the pathways, genes, and specific immune cell types involved using powerful and high-resolution single-cell sequencing (Aims 1 and 2), disease models in mice (Aim 2), rigorous and specific clustered regularly interspaced short palindromic repeats (CRISPR) gene perturbation experiments (Aim 1), and a sensitive protein antigen array (Aim 3). This work will be performed in a world-class environment at Stanford University under the supervision of Dr. Howard Y. Chang, a lncRNA authority who excels at developing and applying sequencing techniques to study diseases, with co-mentorship from Dr. PJ Utz, a clinically trained rheumatologist with particular expertise in autoantibodies and autoimmune disease mouse models. An advisory committee and consisting of experts in single sequencing in immune cells (Dr. Satpathy), high throughput CRISPR screens (Dr. Bassik), proteomics (Dr. Lundberg) and autoimmunity (Dr. Fiorentino) will provide additional mentorship to Dr. Dou and the resources necessary to achieve her project goals. Completing the project and associated training plan will allow Dr. Dou to meet key milestones in her transition to independent investigator. The 3 Aims are designed for parallel investigations. The first half of each aim will be completed during the mentored K99 phase to provide the platform for the R00 independent phase. This project will be the springboard for Dr. Dou to launch an independent career and achieve her long-term goal of resolving the genetic and epigenetic factors underlying autoimmune diseases.

项目概要/摘要 自身免疫性疾病 (AD) 是美国第三大流行疾病，对健康的影响尤为严重 尽管患病率很高且社会成本很高，但有效的治疗方法却很少。 由于自身免疫的遗传因素和环境触发因素仍然很差，因此没有明确的治疗方法 现有研究涉及荷尔蒙差异来解释自身免疫和性别差异。 免疫反应无法调和克莱因费尔特男性对自身免疫性疾病的易感性增加，他们 与女性 (XX) 一样，有两条 X 染色体 (XXY)，但与大多数男性 (XY) 不同，在 XX 个体中，长染色体。 非编码 RNA (lncRNA)、XIST（小鼠中的 Xist）是沉默 X 之一以实现基因剂量所必需的 该项目旨在阐明 XX 相关的自身免疫性疾病发展的优势。 通过研究女性特异性 lncRNA、XIST 以及 XIST 中与 XIST 相关的蛋白质 核糖核蛋白复合物（RNP）作为自身免疫的潜在免疫复合物触发因素。 在此提案中，窦博士提议检验 Xist RNP 增加自身免疫风险的新假设 使用三个多层次目标：(1) 在体外，通过使用 Xist RNP 对免疫细胞进行受控刺激，(2) 在体内 体内，通过转基因 Xist 小鼠的自身免疫性疾病模型，该小鼠携带可表达 Xist 的雄性小鼠 RNP，(3) 诊断性地，使用自身免疫性疾病患者血清来测试针对 XIST RNP 蛋白的反应性。 项目完成将构建通路、基因、特异性免疫细胞的综合模型 涉及使用强大且高分辨率的单细胞测序（目标 1 和 2）、小鼠疾病模型的类型 （目标2），严格且特异性的成簇规则间隔短回文重复（CRISPR）基因 微扰实验（目标 1）和敏感蛋白抗原阵列（目标 3）。 这项工作将在斯坦福大学的世界一流环境中进行，并在以下人员的监督下进行： Howard Y. Chang 博士，lncRNA 权威，擅长开发和应用测序技术 在 PJ Utz 博士的共同指导下研究疾病，PJ Utz 博士是一位经过临床培训、具有特殊专业知识的风湿病学家 自身抗体和自身免疫性疾病小鼠模型的咨询委员会，由专家组成。 免疫细胞单次测序（Satpathy 博士）、高通量 CRISPR 筛选（Bassik 博士）、蛋白质组学 （Lundberg 博士）和自身免疫学（Fiorentino 博士）将为窦博士提供额外的指导和资源 完成该项目和相关培训计划将使窦博士能够 实现她向独立调查员过渡的关键里程碑 这 3 个目标是为并行而设计的。 每个目标的前半部分将在指导的 K99 阶段完成，以提供平台。 该项目将成为窦博士开启独立事业的跳板。 并实现解决自身免疫性疾病的遗传和表观遗传因素的长期目标。