"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)是美国第三大流行疾病，其影响不成比例 女性是男性的4倍。尽管发病率高，社会成本高，但有效的治疗方法寥寥无几。 由于自身免疫的遗传因素和环境触发因素仍然很差，因此没有确定的治疗方法 已定义。现有研究表明荷尔蒙的差异可以解释自身免疫的性别差异和 免疫反应未能解决Kleinfelter男性自身免疫性疾病易感性增加的问题， 有两条X染色体(XXY)，像女性(XX)，不像大多数男性(XY)。在XX个人中，长的 非编码RNA(LncRNA)，XIST(小鼠中的Xist)，是沉默X的其中一个以实现基因剂量所必需的 补偿。本项目旨在阐明XX基因在自身免疫性疾病发病中的优势。 通过对XIST中雌性特异的lncRNA、XIST以及与XIST相关的蛋白质的研究 核糖核蛋白复合体(RNP)作为一种潜在的免疫复合体触发自身免疫。 在这项提案中，窦博士建议检验Xist RNPs增加自身免疫风险这一新假说 使用三个多水平的目标：(1)在体外，通过Xist RNPs对免疫细胞的控制刺激，(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，其中雄性小鼠可见XIST表达 RNPs，(3)诊断性，使用自身免疫病患者血清测试对XIST RNP蛋白的反应性。 该项目的完成将建立一个途径、基因和特定免疫细胞的全面模型 使用强大和高分辨率单细胞测序的类型(目标1和2)，小鼠疾病模型 (目标2)，严谨和特异的规则间隔短回文重复序列(CRISPR)基因 扰动实验(目标1)和敏感的蛋白质抗原阵列(目标3)。 这项工作将在斯坦福大学世界级的环境中进行，由 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博士是一位受过临床培训的具有特殊专业知识的风湿病专家 在自身抗体和自身免疫性疾病小鼠模型中。咨询委员会，由以下方面的专家组成 免疫细胞中的单一测序(萨蒂希博士)、高通量CRISPR筛查(巴西克博士)、蛋白质组学 (Lundberg博士)和自身免疫(Fiorentino博士)将为Dou博士和资源提供额外的指导 是实现她的项目目标所必需的。完成该项目和相关的培训计划将使窦博士能够 满足她向独立调查员过渡的关键里程碑。这三个目标是并行设计的 调查。每个目标的前半部分将在指导K99阶段期间完成，以提供平台 用于R00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 并实现她解决自身免疫性疾病背后的遗传和表观遗传因素的长期目标。