Cellular and Genetic Basis of Systemic Lupus

系统性狼疮的细胞和遗传基础

基本信息

批准号：
9118614
负责人：
Shu Man Fu
金额：
$ 57.98万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-09-28 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9118614
关键词：
Acute Acute Glomerulonephritis Adverse effects Affect Age Antibodies Antigen-Antibody Complex Apoptosis Autoimmune Process Autophagocytosis Biological Markers CRISPR/Cas technology Candidate Disease Gene Cell Line Cells Chromosomes, Human, Pair 1 Chromosomes, Human, Pair 17 Chromosomes, Human, Pair 4 Chronic Glomerulonephritis Clinical Clustered Regularly Interspaced Short Palindromic Repeats Complement Activation Complex Complication Data Deposition Diagnosis Dialysis procedure Disease Disease Progression Embryo End stage renal failure Family Family Sizes Female Future Gene Deletion Gene Order Gene Targeting Genes Genetic Genetic Markers Genetic Predisposition to Disease Genetic study Glomerulonephritis Human Human Chromosomes In Vitro Incidence Kidney Kidney Diseases Kidney Failure Lupus Lupus Nephritis Modeling Morbidity - disease rate Mus Nephritis Nuclear Antigens Organ Pathogenesis Pathway interactions Patients Phenotype Play Predisposition Production Recombinants Renal function Research Resistance Risk Risk Management Role Sample Size Sampling Staging Subgroup Systemic Lupus Erythematosus Translating Transplantation Tubular formation Variant Woman base body system congenic disorder risk ds-DNA genetic analysis genome sequencing lupus prone mice member mitochondrial metabolism mortality mouse model mutant novel patient population podocyte prognostic value programs public health relevance resistant strain sex systemic autoimmune disease whole genome

项目摘要

DESCRIPTION (provided by applicant): Systemic Lupus Erythematosus (SLE), a systemic autoimmune disorder, affects multiple organ systems resulting in significant morbidity and mortality. Lupus nephritis is a devastating complication of SLE, with an increased mortality and risk of progressive renal damage leading to end-stage renal disease (ESRD). To date, there remains no definitive pathway to identify the population of patients with lupus nephritis who are ultimately destined to ESRD. Identification of genes predicting ESRD risk in SLE would permit more vigorous treatment in this subgroup despite the potentially significant side effects. In our study of the genetics of lupus- prone mouse model NZM2328, we have identified separate genes controlling ANA and anti-dsDNA Ab production (Adaz1 on chromosome 4), and controlling both acute and chronic glomerulonephritis (GN) (Cgnz1 on chromosome 1 for controlling chronic GN and Agnz1 on chromosome 1, H-2 complex and Agnz2 on chromosome 17 for controlling acute GN). The phenotypes of NZM2328.Lc1 (Lc1) and Lc1 intrachromosomal recombinant congenic lines and NZM2328.Lc4 confirm our genetic analysis. A novel model for pathogenesis of SLE was proposed in which the genes contributing to lupus are classified into two classes - those affecting autoimmune responsiveness and those affecting end organ damage. These two classes of genes interact, resulting in clinical disease. The concept of genes controlling end organ resistance in SLE is novel. Additional data show that acute and chronic GN are under separate genetic control. A 1.34MB region on mouse chromosome 1 (the Cgnz1 locus) has been shown to contain gene(s) that confer resistance to the progression of acute GN (aGN) to chronic GN (cGN). In this competitive renewal application we wish to continue our research program to identify Cgnz1, the gene(s) within the 1.34Mb regions that confers end organ (kidney) susceptibility to damage. Three specific aims are proposed: Specific Aim 1: To generate podocytes and tubular cell lines from NZM2328 and NZM2328.Lc1R27 (R27) to carryout in vitro gene deletion by the CRISPR/Cas method to identify candidate gene(s) that confer(s) resistance to apoptosis or play an important role in autophagy. Both podocytes and tubular cells will be used. These candidate genes will be targeted in Specific Aim 2; Specific Aim 2: To generate mutant lines with deficiency in targeted genes in (NZM2328XR27)F1 embryos by CRISPR/Cas and to generate homozygous mutants to validate susceptibility to end stage renal damage and to generate podocyte specific deletion mutants to demonstrate the gene(s) that exert(s) effects on podocytes; and Specific Aim 3: To identify candidate gene(s) on the human Cgnz1 1.6Mb locus (Hu Cgnz1) that determine the progression of progressive GN in lupus to ESRD. The results of the proposed studies will be validated and refined in future studies involving larger samples. We anticipate that they may provide biomarkers that are of prognostic value and utility for ESRD risk management. Thus the application will generate significant information regarding the pathogenesis of lupus GN and will have significant translational potentials.

描述（由申请人提供）：系统性红斑狼疮 (SLE) 是一种系统性自身免疫性疾病，影响多个器官系统，导致显着的发病率和死亡率。狼疮性肾炎是 SLE 的一种破坏性并发症，导致死亡率增加和进行性肾损伤的风险。迄今为止，尚无确定的途径来确定最终注定会发生 ESRD 的狼疮性肾炎患者群体。尽管存在潜在的显着副作用，但预测 SLE 中 ESRD 风险的基因的鉴定将允许对该亚组进行更强有力的治疗。在我们对狼疮易发小鼠模型 NZM2328 的遗传学研究中，我们已经鉴定了控制 ANA 和抗 dsDNA Ab 产生的单独基因。（Adaz1 位于 4 号染色体上），并控制急性和慢性肾小球肾炎 (GN)（1 号染色体上的 Cgnz1 用于控制慢性 GN，Agnz1 位于染色体上） 1、H-2 复合物和 17 号染色体上的 Agnz2 用于控制急性 GN） NZM2328.Lc1 (Lc1) 和 Lc1 染色体内重组同源系和 NZM2328.Lc4 的表型证实了我们提出的 SLE 发病机制的新模型。其中导致狼疮的基因分为两类——影响自身免疫反应的基因和影响终末器官损伤的基因。控制 SLE 终末器官抵抗的基因的概念是新颖的。其他数据表明，急性和慢性 GN 受到小鼠 1 号染色体上 1.34MB 的区域（Cgnz1 基因座）的单独控制。已被证明含有能够抵抗急性肾小球肾炎 (aGN) 向慢性肾小球肾炎 (cGN) 进展的基因，在这一竞争性更新应用中，我们希望继续我们的研究计划，以识别其中的基因 Cgnz1。提出了三个具体目标：具体目标 1：从 NZM2328 和 NZM2328.Lc1R27 (R27) 生成足细胞和管状细胞系，以通过 CRISPR 进行体外基因删除。 /Cas 方法来鉴定赋予细胞凋亡抗性或在自噬中发挥重要作用的候选基因。将在特定目标 2 中使用这些候选基因；特定目标 2：通过 CRISPR/Cas 在 (NZM2328XR27)F1 胚胎中生成目标基因缺陷的突变体系，并生成纯合突变体以验证敏感性。终末期肾损伤并产生足细胞特异性缺失突变体以证明对足细胞产生影响的基因；以及具体目标 3：确定人类 Cgnz1 1.6Mb 基因座 (Hu Cgnz1) 上的候选基因，该基因决定狼疮中进展性 GN 进展为 ESRD。我们预计，拟议研究的结果将在未来涉及更大样本的研究中得到验证和完善。它们可以为 ESRD 风险管理提供具有预后价值和实用性的生物标志物，因此该应用程序将产生有关狼疮 GN 发病机制的重要信息，并将具有重大的转化潜力。