Complement C3 is Essential for Development of Kidney Cysts in ADPKD Mice

补体 C3 对于 ADPKD 小鼠肾囊肿的发育至关重要

• 批准号: 8512389
• 负责人: Tianqing Kong
• 金额: $ 7.9万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8512389
• 关键词: Affect; Age; Antibodies; Autosomal Dominant Polycystic Kidney; Biology; C3 gene; Cell Line; Cells; Chronic; Chronic Kidney Failure; Cleaved cell; Clinical; Complement; Complement 3; Cyst; Cystic kidney; Development; Dialysis procedure; Disease; End stage renal failure; Enzyme-Linked Immunosorbent Assay; Gene Deletion; Genes; Genetic Polymorphism; Genotype; Growth Factor; Hereditary Disease; Histology; Human; Immunology; In Situ Hybridization; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Inherited; Interleukin-6; Kidney; Knock-out; Knockout Mice; Laboratories; Life; Liquid substance; Literature; Live Birth; Maine; Mass Spectrum Analysis; Mediator of activation protein; Mus; Mutation; Outcome Study; PKD2 gene; Patients; Play; Prevalence; Primary Cell Cultures; Process; Production; Proteins; Publishing; Quality of life; Reaction; Renal Replacement Therapy; Renal function; Reporting; Research; Research Project Grants; Role; Severities; Staging; Structure; Symptoms; Testing; Therapeutic; Time; Tissues; Tumor Necrosis Factor-alpha; Western Blotting; Work; animal facility; base; cell type; cytokine; expectation; human TNF protein; improved; in vivo; kidney epithelial cell; mouse model; novel; novel strategies; novel therapeutics; public health relevance; research study

DESCRIPTION (provided by applicant): This proposal is to determine whether complement C3 is required for development and progressive expansion of kidney cysts induced by Pkd1 inactivation in mice. As the most common inherited disease, autosomal dominant polycystic kidney disease (ADPKD) affects approximately 1 in 400 to 1000 live births. Approximately 85% of ADPKD results from mutation of PKD1, and the remaining 15% are from mutation of PKD2 gene. It is the progressive expansion and enlargement of numerous cysts that destroys normal kidney structure, impairs kidney function and causes clinical symptoms. There are also other factors that control onset and severity of ADPKD, especially aggressive progression of kidney cysts. Enlargement of kidney cysts is a slowly inflammatory process. Cytokines, growth factors and other inflammatory mediators are involved, but the exact mechanism and key mediators remain unclear. We found that complement C3 was elevated and activated in mouse and human AKPKD kidneys and cystic fluid. The local concentration and activation of complement C3 may be involved in development and expansion of kidney cysts. Based on the literature, complement C3 is one of the most highly induced genes in kidney tissue from a PKD mouse model. It is also present and highly expressed in cystic fluid and kidney tissue in human ADPKD. Thus, in order to further determine the essential role of complement C3 in development and aggressive expansion of kidney cysts in vivo, we will generate a double knockout mice (Mx1Cre+Pkd1flox/flox C3-/-) in which the C3 gene is deleted, and Pkd1 can also be conditionally knocked-out by crossing conditional ADPKD mice (Mx1Cre+Pkd1flox/flox) with C3 knockout mice (C3-/-). We will carry out several experiments to investigate how complement C3 promotes the aggressive cystic development, which includes the cell type producing local increased C3, the role of activated and cleaved fragments of C3 leading to focal inflammation and cystic expansion, the mechanism of C3 activation in cystic kidney (Aim 1). In mouse models, we expect that deletion of the complement C3 gene blocks or delays development and progression of kidney cysts induced by Pkd1 inactivation (Aim 2). Approximately one-half of ADPKD patients progress to chronic renal failure by age 60. Current treatment for this disease is dialysis or renal replacement therapy. Our proposed study would reveal a novel therapeutic strategy for this most common life-threatening genetic disease.

描述(由申请人提供)：这项建议是为了确定补体C3是否在由PKD1失活引起的小鼠肾囊肿的发展和进行性扩张中所必需。常染色体显性遗传性多囊肾病(ADPKD)是最常见的遗传性疾病，约占活产儿的1/400~1/1000。约85%的ADPKD是由PKD1基因突变引起的，其余15%是由PKD2基因突变引起的。它是大量的囊肿进行性扩张和增大，破坏正常的肾脏结构，损害肾功能，并导致临床症状。还有其他因素控制ADPKD的发病和严重程度，特别是肾囊肿的侵袭性进展。肾囊肿大是一个缓慢的炎症过程。细胞因子、生长因子和其他炎症介质参与其中，但具体机制和关键介质尚不清楚。我们发现补体C3在小鼠和人AKPKD肾脏和囊液中升高并被激活。补体C3的局部聚集和激活可能参与了肾囊肿的发生发展。根据文献，补体C3是PKD小鼠模型肾组织中诱导作用最强的基因之一。在ADPKD患者的囊液和肾组织中也有高表达。因此，为了进一步确定补体C3在体内肾囊肿发育和侵袭性扩张中的重要作用，我们将构建一个C3基因缺失的双基因敲除小鼠(Mx1Cre+Pkd1flx/Flox C3-/-)，并将条件性ADPKD小鼠(Mx1Cre+Pkd1flx/Flox)与C3基因敲除小鼠(C3-/-)杂交，也可以有条件地敲除PKD1。我们将开展几个实验来研究补体C3如何促进囊性侵袭性发育，包括产生局部增加的C3的细胞类型，C3的激活和裂解片段导致局灶性炎症和囊性扩张的作用，以及C3在囊性肾脏中的激活机制(目标1)。在小鼠模型中，我们预计补体C3基因的缺失阻止或延缓了由PKD1失活引起的肾囊肿的发展和进展(目标2)。大约一半的ADPKD患者在60岁前进展为慢性肾功能衰竭。目前治疗这种疾病的方法是透析或肾脏替代疗法。我们提议的研究将揭示一种新的治疗策略，以治疗这种最常见的威胁生命的遗传病。