Cellular and Genetic Basis of Systemic Lupus

描述（由申请人提供）： 系统性红斑狼疮（SLE）是一种多系统疾病，发病率和死亡率高，主要影响育龄女性。遗传和环境因素在其发病中起重要作用。尽管取得了重大进展，但免疫介导的肾小球肾炎（GN）仍然是终末期肾衰竭的主要原因。PI的实验室通过研究小鼠模型NZM 2328专注于狼疮GN的遗传方面。这项竞争性更新申请是为了继续PI的研究计划，以确定1号染色体上的狼疮易感基因Cgnz 1和Adnz 2，它们分别在NZM 2328（一种研究较多的人类增殖性狼疮肾炎模型）中产生抗核抗体和相关自身抗体（Ab），并对慢性肾小球肾炎（cGN）的发展产生终末器官抗性。在本研究期间，获得了两个信息丰富的同源系290-2和507-2。该分析得出的结论是，该区域已缩小到1.34 Mb的区域，这非常适合进一步分析。自身免疫和终末器官损伤的分离是独特的，PI的实验室仍然是关注有助于终末器官抵抗损伤的基因的主要场所。一个具有互补技能的调查人员团队已经组建，以利用分子和细胞生物学以及动物遗传学的最新技术来解决这一技术要求很高的研究项目。为了实现本项目的长期目标，提出了四个具体目标：具体目标1：产生和表征NZM.C57Lc1重组同源菌株Lc 1（1.34Mb），其含有感兴趣的1.34Mb（NZM2328X507-2）F1 X（NZM 2328 X507 -2）F1;特定目标2：为了从NZM 2328 BAC文库中鉴定一组连续重叠的基因组克隆（重叠群），其覆盖了1.34Mb的感兴趣区域，并对这些重叠群进行测序，以通过将这些序列与数据库中可获得的129和B6/C57 L的序列进行比较来确定该区域中的多态性。多态性还可以鉴定Cgnz 1和Adaz 2的潜在候选基因;特异性目的3：确定该区域内45个基因在固有肾细胞和免疫细胞（包括T和B细胞、树突细胞、单核细胞和巨噬细胞以及NK细胞）中的转录谱，以鉴定负责肾抗损伤和负责增强自身免疫应答和特异性目的4的候选基因：通过基因敲入技术或等位基因转基因验证易感性。结果将提供确凿的证据表明，控制终末器官损伤的基因在狼疮肾炎中起着重要作用，它们与增强自身免疫的基因相互作用，导致各种临床表现。在基本层面上，这些结果可能会改变我们对自身免疫性疾病发病机制的看法。此外，这些结果具有重要的临床意义，因为遗传因素可能使我们能够定制狼疮肾炎的个体治疗方案，以最大限度地提高治疗目标并最大限度地减少毒副作用。

DESCRIPTION (provided by applicant): Systemic lupus erythematosus (SLE), a multi-system disorder with significant morbidity and mortality affects predominantly females in their reproductive years. Genetic and environmental factors play significant roles in its pathogenesis. Despite significant progress, immune mediated glomerulonephritis (GN) remains a major cause of end stage renal failure. The PI's laboratory has been focused on the genetic aspect of lupus GN by studying the murine model NZM2328. This competitive renewal application is to continue the PI's research program to identify lupus susceptibility genes, Cgnz1 and Adnz2, on chromosome 1 that confer the production of antinuclear and related autoantibodies (Ab) and end organ resistance to the development of chronic glomerulonephritis (cGN) respectively in NZM2328, a much studied model for human proliferative lupus nephritis. During the currrent grant period, two informative congenic lines, 290-2 and 507-2 were generated and characterized. This analysis led to the conclusion that this region has been narrowed to a 1.34Mb area that is very much amendable to further analysis. The separation of autoimmunity and end organ damage is unique and the PI's laboratory remains the major site to focus on the genes which contribute to end organ resistance to damage. A team of investigators with complementary skills have been assembled to utilize up-to- date techniques in molecular and cell biology and animal genetics to resolve this technically demanding research project. To achieve the long term objective of this program, four specific aims are proposed: Specific Aim 1: To generate and to characterize the NZM.C57Lc1 recombinant congenic strain Lc1(1.34Mb) that contains the 1.34Mb of interest by intercross breeding (NZM2328X507-2)F1 X (NZM2328X507-2)F1; Specific Aim 2: To identify a set of contiguous overlapping genomic clones (contigs) from the NZM2328 BAC library, which cover the 1.34 Mb region of interest and to sequence these contigs to define polymorphisms in this region by comparing the sequences with those of 129 and B6/C57L available in the database. The polymorphisms may also identify potential candidate genes for Cgnz1 and Adaz2; Specific Aim 3: To determine the transcriptional profiles of the 45 genes within this region in intrinsic kidney cells and in immune cells including T and B cells, dendritic cells, monocytes and macrophages and NK cells to identify candidate genes responsible for kidney resistance to damage and for those for enhanced autoimmune response and Specific Aim 4: To validate susceptibility by gene knockin technology or allele transgenesis. The results will provide conclusive evidence that genes controlling end organ damage play an important role in lupus nephritis and that they interact with genes that enhance autoimmunity resulting in various clinical presentations. On the basic level, the results may alter our thinking on the pathogenesis of autoimmune disorders. In addition, these results have significant clinical implication in that genetic factors may allow us to tailor individual therapeutic regimens for lupus nephritis to maximize therapeutic goals and to minimize toxic side effects.