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）：该提案是为了确定小鼠pKD1诱导的肾脏囊肿的发展和逐步扩张是否需要补充C3。作为最常见的遗传疾病，常染色体显性多囊性肾脏疾病（ADPKD）影响了400至1000个活产中约1个。 ADPKD的大约85％是由PKD1突变引起的，其余15％来自PKD2基因的突变。大量囊肿的进行性扩张和扩大会破坏正常的肾脏结构，损害肾脏功能并导致临床症状。还有其他控制ADPKD发作和严重程度的因素，尤其是肾脏囊肿的侵略性进展。肾脏囊肿的扩大是缓慢的炎症过程。涉及细胞因子，生长因子和其他炎症介质，但确切的机制和关键介体仍不清楚。我们发现补体C3在小鼠和人AKPKD肾脏和囊性液中升高并激活。补体C3的局部浓度和激活可能与肾脏囊肿的发育和扩展有关。基于文献，补体C3是PKD小鼠模型中肾脏组织中最高度诱导的基因之一。它也存在于人ADPKD中的囊性液和肾脏组织中。因此，为了进一步确定补体C3在体内肾脏囊肿的发育和积极扩张中的重要作用，我们将产生双基因敲除小鼠（MX1CRE+PKD1FLOX/FLOX C3 - / - ），其中C3基因被删除，并且PKD1也可以通过条件敲除条件敲除条件敲除条件的条件下adpkd meicea adpkd鼠标。 （MX1CRE+PKD1FLOX/FLOX）用C3敲除小鼠（C3 - / - ）。我们将进行几项实验，以研究补体C3如何促进攻击性囊性发育，其中包括产生局部C3的细胞类型，C3的激活和切割片段的作用，导致局灶性炎症和囊性扩张，C3激活机制在囊性肾脏中的C3激活机制（AIM 1）。在小鼠模型中，我们预计补体C3基因块的缺失或延迟了PKD1失活诱导的肾脏囊肿的发育和进展（AIM 2）。到60岁时，大约一半的ADPKD患者会导致慢性肾衰竭。该疾病的当前治疗方法是透析或肾脏替代疗法。我们提出的研究将揭示针对这种最常见的威胁生命的遗传疾病的新型治疗策略。