"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.

项目总结/摘要 自身免疫性疾病（AD）是美国第三大流行病，不成比例地影响着 女性比男性多4倍。尽管患病率高，社会成本高，但有效的治疗方法很少 由于自身免疫的遗传因素和环境触发因素仍然很差， 定义了现有的研究暗示激素差异可以解释自身免疫性的性别差异， 免疫反应不能调和Kleinfelter男性对自身免疫性疾病的易感性增加， 有两个X染色体（XXY），像女性（XX），而不像大多数男性（XY）。在XX个体中，长 非编码RNA（lncRNA），XIST（小鼠中的Xist），是沉默X之一以达到基因剂量所必需的 赔偿这个项目的目的是阐明XX-连锁优势的自身免疫性疾病的发展 通过对XIST中女性特异性lncRNA、XIST及其相关蛋白的研究， 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剂。 在这项提案中，Dou博士提出测试Xist RNP增加自身免疫风险的新假设。 使用三个多层次的目标：（1）在体外，通过用Xist RNP控制刺激免疫细胞，（2）在 体内，通过在转基因Xist小鼠中建立自身免疫性疾病模型，其中雄性小鼠可存活地表达Xist RNP，（3）诊断上，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模型的途径，基因和特定的免疫细胞 使用强大的高分辨率单细胞测序（目的1和2），小鼠疾病模型 (Aim 2）、严谨特异的成簇规则间隔短回文重复序列（CRISPR）基因 微扰实验（Aim 1）和敏感蛋白抗原阵列（Aim 3）。 这项工作将在斯坦福大学的世界级环境中进行， 博士霍华德Chang是lncRNA权威，擅长开发和应用测序技术， 研究疾病，与PJ乌茨博士共同指导，他是一位经过临床训练的风湿病学家，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。一个咨询委员会，由以下方面的专家组成： 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 (Dr. Lundberg）和自身免疫（Fiorentino博士）将为Dou博士提供额外的指导和资源 以实现其项目目标。完成项目和相关培训计划将使Dou博士能够 完成她向独立调查员过渡的关键里程碑。三个目标旨在并行 调查事务所每个目标的前半部分将在辅导K99阶段完成，以提供平台 对于R 00独立阶段。这个项目将成为窦博士开始独立职业生涯的跳板 并实现她的长期目标，解决自身免疫性疾病的遗传和表观遗传因素。