Genetic Programming in Progressive Renal Disease

进行性肾病的基因编程

• 批准号: 8010742
• 负责人: Erwin P. Bottinger
• 金额: $ 10.68万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8010742
• 关键词: Adaptor Signaling Protein; Apoptosis; Binding; CD2-associated protein; Cell Survival; Complex; Diabetic Nephropathy; Diabetic mouse; Disease susceptibility; Down-Regulation; Doxycycline; Epithelial; Epithelial Cells; Genetic Programming; Genotype; Grant; Human; In Vitro; Injury; Kidney; Kidney Diseases; Knockout Mice; Lesion; Life; Ligands; Mediating; Modeling; Molecular; Mus; Mutation; Parietal; Pathway interactions; Phosphatidylinositols; Phosphotransferases; Receptor Signaling; Regulation; Renal glomerular disease; Role; Signal Transduction; Structural Models; TGF-beta type I receptor; Technology; Testing; Transgenes; Transgenic Organisms; Tubular formation; Up-Regulation; Work; base; diabetic; glomerulosclerosis; in vivo; men; mouse model; novel; podocyte; prevent; protein complex; protein expression; protein function; receptor; slit diaphragm

Podocyte depletion is a hallmark of progressive glomerulosclerosis and diabetic nephropathy. CD2- associated protein (CD2AP) is a widely-expressed cytoplasmic adaptor protein that has previously been implicated in podocyte slit diaphragm protein complex formation. CD2AP null mice develop progressive glomerulosclerosis early in life and CD2AP haploinsufficiency is associated with glomerular disease susceptibility in humans. However, it remains unclear how loss of CD2AP function may cause glomerulosclerosis. In this renewal application, we will extend our work to test the hypothesis that CD2AP promotes renal epithelial cell survival by functioning as a molecular switch that suppresses the TGF-beta receptor type I (Tgfbr1)/Smad3-dependent apoptosis pathway and activates a Tgfbrl/phosphoinositide-3- kinase (PI3K) -dependent survival pathway. If confirmed, our studies will provide a molecular mechanism to explain how loss of CD2AP may enhance podocyte apoptosis and depletion to promote glomerular disease in mice and men based on a novel, essential functional role for CD2AP as multi-pathway regulator in TbRI- induced signal transduction. The relevance of this work is further underscored by recent observations suggesting that in the absence of direct genetic defects, functional downregulation of CD2AP expression is a common finding in mouse models and human glomerular disease. Specific Aims: 1. Identify the molecular determinants and characterize the regulation of the TGF-p-inducible interaction of CD2AP and cytoplasmic Tgfbrl. We will a) determine the sequence motif and structural model of Tgfbrl that underlies the interaction with CD2AP, and b) compare it with the Smad-binding model mediated by the GS region and L45 loop of Tgfbrl. 2. Investigate how CD2AP mediates negative regulation of Tgfbr1/Smad3 signaling. We will examine whether a) CD2AP competes with SmadS for binding to activated Tgfbrl; and b) CD2AP targets internalized Tgfbrl complexes for degradation and termination of signaling. 3. Define the role of podocyte-specific TGF-b receptor signaling in podocyte apoptosis and depletion in vivo, and determine whether this podocyte-specific mechanism underlies the progression of glomerular injury to glomerulosclerosis in CD2AP-/- mice? We will determine in vivo whether a) conditional deletion of TGF-b receptor type II (Tgfbr2) in podocytes using cre/lox technology prevents podocyte apoptosis, depletion, and/or glomerulosclerosis in CD2AP-/- mice; and b) whether podocyte-specific, controlled activation of TGF-(3 receptor signaling is sufficient to induce podocyte apoptosis, podocyte depletion and glomerulosclerosis in vivo using doxycycline-inducible expression of constitutively active Tgfbrl (Tgfbrl (AAD)).

足细胞耗竭是进行性肾小球硬化和糖尿病性肾病的标志。 CD2- 相关蛋白（CD2AP）是一种表达广泛的细胞质衔接蛋白，以前曾经是 与足细胞裂缝隔膜蛋白复合物形成有关。 CD2AP无效小鼠发展进步 肾小球硬化的生命早期和CD2AP单倍体不足与肾小球疾病有关 人类的敏感性。但是，尚不清楚CD2AP功能的损失可能导致 肾小球硬化。在此续订应用中，我们将扩展工作以测试CD2AP的假设 通过充当抑制TGF-beta的分子开关来促进肾上皮细胞存活 受体I型（TGFBR1）/SMAD3依赖性凋亡途径，并激活TGFBRL/phosphoinositide-3- 激酶（PI3K）依赖性生存途径。如果得到证实，我们的研究将提供分子机制 解释CD2AP的损失如何增强足细胞凋亡和耗竭以促进肾小球疾病 在小鼠和男性中，基于新颖的，CD2AP作为TBRI-中的多条纹调节剂的新型功能作用 诱导信号转导。最近的观察进一步强调了这项工作的相关性 表明在没有直接遗传缺陷的情况下，CD2AP表达的功能下调是A 在小鼠模型和人肾小球疾病中的共同发现。 具体目的：1。识别分子决定因素并表征TGF-P-P-P-P-P-P-P的调节 CD2AP和细胞质TGFBRL的相互作用。我们将a）确定序列基序和结构模型 与CD2AP相互作用的基础的TGFBRL，b）将其与SMAD结合模型进行比较 由TGFBRL的GS区域和L45环介导。 2。研究CD2AP如何介导负调节 TGFBR1/SMAD3信号传导。我们将检查A）CD2AP是否与SMADS竞争以结合到 活化的TGFBRL; b）CD2AP靶向内部化的TGFBRL复合物，以降解和终止 信号。 3。定义足细胞特异性TGF-B受体信号传导在足细胞凋亡和 体内耗尽 CD2AP - / - 小鼠肾小球硬化的肾小球损伤？我们将在体内确定a）条件是否有条件 使用CRE/LOX技术在足细胞中删除TGF-B受体II型（TGFBR2） CD2AP - / - 小鼠中的凋亡，耗竭和/或肾小球硬化； b）是否适合Podocyte特异性 TGF-的受控激活（3个受体信号传导足以诱导足细胞凋亡，足细胞 使用多西环素诱导的组成性活性表达在体内耗竭和肾小球硬化 TGFBRL（TGFBRL（AAD））。

项目成果

Smad2 and myocardin-related transcription factor B cooperatively regulate vascular smooth muscle differentiation from neural crest cells.

• DOI: 10.1161/circresaha.113.301921
• 发表时间: 2013-09-27
• 期刊: Circulation research
• 影响因子: 20.1
• 作者: Xie WB;Li Z;Shi N;Guo X;Tang J;Ju W;Han J;Liu T;Bottinger EP;Chai Y;Jose PA;Chen SY
• 通讯作者: Chen SY

The molecular and functional phenotype of glomerular podocytes reveals key features of contractile smooth muscle cells.

• DOI: 10.1152/ajprenal.00559.2007
• 发表时间: 2008-10
• 期刊: American journal of physiology. Renal physiology
• 作者: Saleem MA;Zavadil J;Bailly M;McGee K;Witherden IR;Pavenstadt H;Hsu H;Sanday J;Satchell SC;Lennon R;Ni L;Bottinger EP;Mundel P;Mathieson PW
• 通讯作者: Mathieson PW