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，发病率和死亡率显著，主要影响育龄女性。遗传和环境因素在其发病机制中起重要作用。尽管取得了重大进展，但免疫介导性肾小球肾炎(GN)仍然是终末期肾功能衰竭的主要原因。PI的实验室通过研究小鼠模型NZM2328，专注于狼疮GN的遗传方面。这一竞争性更新应用是为了继续PI的研究计划，以确定1号染色体上的狼疮易感基因Cgnz1和Adnz2，这些基因分别在NZM2328中产生抗核抗体(Ab)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抵抗。NZM2328是一种广泛研究的人类增殖性狼疮性肾炎模型。在目前的授权期内，产生了两个信息丰富的同源系290-2和507-2，并对其进行了鉴定。这一分析得出的结论是，该区域已缩小到1.34Mb的区域，这一区域非常值得进一步分析。自身免疫和终末器官损伤的分离是独一无二的，PI的实验室仍然是专注于促进终末器官抵抗损伤的基因的主要场所。一组具有互补技能的研究人员已经聚集在一起，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高的研究项目。为了实现该计划的长期目标，提出了四个具体目标：具体目标1：通过杂交育种(NZM2328X507-2)F1 X(NZM2328X507-2)F1，产生并鉴定包含1.34Mb的NZM.C57Lc1重组同源菌株LC1(1.34Mb)；特定目标2：从NZM2328BAC文库中鉴定一组连续重叠的基因组克隆(重叠)，覆盖1.34Mb的感兴趣区域，并对这些重叠序列进行测序，通过与数据库中现有的129和B6/C57L的序列比较来确定该区域的多态性。这些多态性还可能识别Cgnz1和Adaz2的潜在候选基因；特定目的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、树突状细胞、单核细胞和巨噬细胞以及NK细胞)中的转录谱，以确定与肾脏抵抗损伤和增强自身免疫反应有关的候选基因，以及特定目的4：验证基因敲击技术或等位基因转基因的易感性。这些结果将提供确凿的证据，证明控制终末器官损害的基因在狼疮性肾炎中发挥重要作用，并且它们与增强自身免疫的基因相互作用，从而导致不同的临床表现。从根本上讲，这一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，因为遗传因素可能允许我们为狼疮性肾炎量身定做个性化的治疗方案，以最大限度地实现治疗目标，并将毒副作用降至最低。 公共卫生相关性： 系统性红斑狼疮是一种影响许多器官的疾病。它造成了大量的痛苦和早逝。它主要影响处于生育年龄的女性。遗传因素起着重要作用。尽管研究取得了重大进展，但肾脏疾病仍然是终末期肾功能衰竭的主要原因，需要进行长期透析或肾移植。这项提议是为了寻求资金来继续一项研究计划，以确定影响自身免疫的遗传因素，并使肾脏抵抗自身抗体和/或自身反应性淋巴细胞的损害。该研究计划利用了在研究人员实验室建立的独特的小鼠模型。研究计划的结果可以很容易地转化为患者护理，因为个别患者可以使用单独的方案治疗他们的肾脏疾病，并保持他们的病情缓解。这将显著降低发病率和死亡率，使患者能够保持活跃和多产。因此，研究计划应该对公共健康产生重大影响。

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. PUBLIC HEALTH RELEVANCE: Systemic lupus erythematosus is a disease affecting many organs. It causes a significant amount of suffering and early death. It affects predominantly females at their reproductive ages. Genetic factors play a significant role. Despite significant progress in research, renal disease remains a major cause of end stage renal failure that requires either chronic dialysis or renal transplantation. This proposal is to seek funding to continue a research program to identify genetic factors that affect autoimmunity and render the kidney resistant to damage by autoantibodies and/or autoreactive lymphocytes. The research program is taking advantage of the unique mouse models established in the investigators' laboratory. The results of the research program can be readily translated to patient care in that individual patients can be treated with individual regimens for their renal disease and to maintain them in remission. This will reduce the morbidity and mortality significantly so that patients can remain active and productive. Thus the research program should have significant impact on public health.