"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: 10525045
• 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/10525045
• 关键词: Address; Advisory Committees; Affect; American; Americas; Antigen-Antibody Complex; Antigens; Atlases; 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; Data; 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; Immunization; In Vitro; Individual; Investigation; Klinefelter' s Syndrome; Link; Malignant Neoplasms; Mentors; Mentorship; Modeling; Monitor; Mouse Protein; 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; Testing; Tissues; Training; Transgenic Mice; Transgenic Organisms; Universities; Untranslated RNA; Women' s Health; Work; X Chromosome; X Inactivation; XX male; authority; autoreactivity; biological sex; career; cell type; cost; design; effective therapy; epigenome; experimental study; human disease; immune activation; in vivo; innovation; male; mouse model; multidisciplinary; 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.

项目摘要 /摘要 自身免疫性疾病（AD）是美国第三大流行的疾病，影响不成比例 女性比男性高4倍。尽管社会的流行率很高和成本，但有效的治疗很少 而且没有确定的治疗，因为自身免疫的遗传因素和环境触发因素仍然很差 定义。现有的研究暗示了马差异，以解释自身免疫性的性别差异和 免疫反应无法调和Kleinfelter雄性自身免疫性疾病的易感性增加，他们 具有两个X染色体（XXY），例如女性（XX），与大多数男性（XY）不同。在XX个人中，长期 沉默的X剂量之一需要实现基因剂量，需要一个非编码RNA（lncRNA），xist（小鼠中的xist）（xist） 赔偿。该项目旨在阐明自身免疫性疾病发展的XX连接优势 通过研究女性特异性lncRNA，xist和与Xist相关的蛋白 核糖核蛋白复合物（RNP）是自身免疫性的潜在免疫复合触发。 在此提案中，Dou博士的提议测试了新的假设，即XIST RNP会增加自身免疫风险 使用三个多层次的目标：（1）在体外，通过用XIST RNP对免疫细胞进行控制刺激，（2） 体内，通过转基因Xist小鼠中的自身免疫性疾病建模，其中雄性小鼠通过表达Xist RNP，（3）诊断，使用自身免疫性疾病患者血清测试针对XIST RNP蛋白的反应性。 该项目的完成将建立一个全面的途径，基因和特定的免疫电池模型 使用强大和高分辨率的单细胞测序涉及的类型（目标1和2），小鼠疾病模型 （AIM 2），严格而特定的定期插入短的短质体重复序列（CRISPR）基因 扰动实验（AIM 1）和敏感的蛋白质抗原阵列（AIM 3）。 这项工作将在斯坦福大学的世界一流环境中进行 霍华德·Y. Chang博士（Lncrna Authority），擅长开发和应用测序技术 研究疾病，由临床训练的风湿病学家PJ Utz博士的冠军制度，具有特殊的专业知识 在自身抗体和自身免疫性疾病小鼠模型中。咨询委员会，由专家组成 免疫细胞中的单个测序（Dr. Satpathy），高通量CRISPR筛选（Bassik博士），蛋白质组学 （Lundberg博士）和自动免疫（Fiorentino博士）将为Dou博士和资源提供额外的精神训练 实现她的项目目标所必需的。完成项目和相关的培训计划将使Dou博士能够 在过渡到独立调查员时，遇到了关键里程碑。 3个目标是为平行设计的 调查。每个目标的前半部分将在修订的K99阶段完成，以提供平台 对于R00独立阶段。该项目将是Dou博士开展独立职业的跳板 并实现她解决自身免疫性疾病的遗传和表观遗传因素的长期目标。