Glomerular Capillary Wall--Normal and Pathologic

肾小球毛细血管壁——正常和病理

基本信息

批准号：
7459903
负责人：
Yashpal S. Kanwar
金额：
$ 36.39万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-04-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7459903
关键词：
Ablation Adult Affect Alleles Animal Model Antigens Binding Biochemical Biology Brassicaceae Breeding Cathepsins Cells Collagen Collagen Type IV Data Development Diabetes Mellitus Diabetic Nephropathy Embryo Evolution Exons Extracellular Matrix Extracellular Matrix Proteins Fibronectins Follistatin Generations Genes Genetic Transcription Genitourinary system Glomerular Capillary Glucose Guanosine Triphosphate Phosphohydrolases HK2 gene Heterozygote Human Glandular Kallikrein 2 Hyperglycemia In Vitro Injury Kidney Knock-out Knockout Mice Laminin Lesion Life Masks Matrix Metalloproteinases Metanephric structure Molecular Morbidity - disease rate Morphology Mus Neonatal Newborn Infant Pathologic Phenotype Protein Isoforms Protein Overexpression Proteins Rattus Reactive Oxygen Species Recombinants Reporting Research Personnel Role Signal Transduction Signaling Molecule Site Staging Stream Streptozocin System Tetanus Helper Peptide Tin Transforming Growth Factor beta Transgenic Mice Tubular formation Tubulointerstitial Nephritis body system db/db mouse diabetic fetal glycation glycosylation homologous recombination in vivo inhibitor/antagonist insight interstitial mRNA Expression mature animal mortality mutant nephrogenesis novel osteopontin procathepsin B progenitor programs promoter protein protein interaction research study treatment duration vector yeast two hybrid system

项目摘要

Diabetes mellitus is a major cause of mortality and morbidity as hyperglycemia adversely affects the biology of various organ systems during embryonic and adult life. Major complications in newborns include congenital anomalies, such as hypo/agenesis of the urogenital system, and diabetic nephropathy in adults. In diabetic nephropathy, both the glomerular and tubulo-interstitial compartments are involved, where an increase of extracellular matrix (ECM) is observed. ECM accumulation is due to increased synthesis of matrix proteins, e.g., collagen and fibronectin, and decreased activity of MMPs, and glycation of ECM proteins with formation of advanced glycation products (AGEs), leading to generation of reactive oxygen species (ROS), which via down-stream signals, such as TGF-beta, induce progressive glomerular and tubulo-interstitial injury. In addition, altered expression of other ECM proteins, e.g., osteopontin and fibronectin, has been reported. However, the role of a recently characterized ECM protein, tubulointerstitial nephritis antigen (TIN-ag), in diabetic nephropathy remains enigmatic. TIN-ag is a mosaic protein containing procathepsin B like domain, follistatin module, ATP binding motif, glycosylation sites and segmental homology with other ECM proteins. It is developmentally regulated, is expressed in progenitor of tubules, and modulates tubulogenesis in vitro. To decipher the role of TIN-ag in progressive renal injury during embryonic and adult life, experiments are proposed under the following specific aims: I. Role of TIN-ag in glucose-induced renal dysmorphogenesis will be investigated since treatment of fetal mice kidneys with high glucose or TIN-ag antisense yield similar phenotype, i.e., inhibited tubulogenesis, suggesting possible relevance of TIN-ag in the evolution of glucose-induced lesions. II. Role of TIN-ag in animal models overexpressing and masking TIN-ag gene will be investigated, using Tet-On/Off systems in transgenic mice and fetal Lewis rats. III. TIN-ag and its domain-specific interactions with other ECM proteins will be assessed since TIN-ag is masked in adult Lewis rats. Recombinant TIN-ag will be generated to study protein:protein interactions with isoforms of type-IV collagen and laminin, and novel interacting partners will be identified by yeast two-hybrid system. IV. Role of TIN-ag in various animal models of diabetic nephropathy, and mechanisms relevant to its increased expression will be investigated by morphologic and biochemical studies. In vitro culture system will be also employed to delineate the mechanisms of altered expression of TIN-ag in diabetic milieu. V. Role of TIN-ag in renal tubulo-interstitial pathobiology will be investigated by generating TIN-ag conditional and conventional knock out (KO) mice. For generation of the conditional KO mice, heterozygote gouty mice will be cross-bred with Ella Cre and then with Ksp Cre mice that would have lesions confined to kidney. Null alleles will be interbred to generate Null -/- mutant as a conventional KO mice with ablation of the TIN-ag gene.

糖尿病是死亡率和发病率的主要原因，因为高血糖在胚胎和成人生活期间对各种器官系统的生物学产生不利影响。新生儿的主要并发症包括先天性异常，例如泌尿生殖系统的低/发育性和成人糖尿病性肾病。在糖尿病性肾病中，涉及肾小球和肾小管间隙室，其中观察到细胞外基质（ECM）的增加。 ECM积累是由于基质蛋白的合成增加，例如胶原蛋白和纤连蛋白，以及 MMPS的活性降低，ECM蛋白的糖素与高级糖基化产物（AGES）的形成，导致活性氧（ROS）产生，通过下游信号（例如TGF-beta），例如TGF-beta，例如诱导进行性肾小球和小管相关性损伤。此外，已经报道了其他ECM蛋白的表达改变，例如骨桥蛋白和纤连蛋白。但是，最近表征的ECM蛋白质TubuloInterstitial的作用糖尿病性肾病中的肾炎抗原（TIN-AG）仍然神秘。 Tin-AG是一种镶嵌蛋白，含有类似域的proathepsin B，Follistatin模块，ATP结合基序，糖基化位点和与其他ECM蛋白的分段同源性。它在发育中受到调节，在小管的祖细胞中表达，并在体外调节小管生成。为了破译锡ag在胚胎和成人生活中进行性肾脏损伤中的作用，在以下具体目的下提出了实验：I。锡Ag在葡萄糖诱导的肾脏肾上腺异常中的作用将是研究以来，由于治疗高葡萄糖或锡抗反义的胎儿肾脏的治疗肾脏会产生相似的表型，即抑制的肾小管生成，表明锡AG在葡萄糖诱导的病变的进化中可能相关性。 ii。将使用转基因小鼠和胎儿路易斯大鼠中的TET-ON/OFF系统研究锡Ag在过表达和掩盖锡Ag基因的动物模型中的作用。 iii。 TIN-AG及其与其他ECM蛋白的域特异性相互作用将是由于在成年刘易斯大鼠中掩盖了锡ag，因此评估。将生成重组TIN-AG来研究蛋白质：与IV型胶原蛋白和层粘连蛋白的同工型的蛋白质相互作用，并且新型相互作用伴侣将通过酵母两杂交系统鉴定。 iv。 Tin-Ag在糖尿病性肾病的各种动物模型中的作用，以及与它的表达增加将通过形态学和生化研究研究。还将采用体外培养系统来描述糖尿病环境中锡Ag表达改变的机制。 V. Tin-ag在肾小管间隙病理生物学中的作用将通过产生条件和常规敲除（KO）小鼠来研究。为了生成条件的KO小鼠，杂合子痛风小鼠将与Ella Cre杂交，然后将其与肾脏限制在肾脏的病变相交。无效等位基因将被杂交以产生零 - / - 突变体作为传统的KO小鼠，并消融了锡 - Ag基因。

项目成果

期刊论文数量（64）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cloning of mouse c-ros renal cDNA, its role in development and relationship to extracellular matrix glycoproteins.

小鼠 c-ros 肾 cDNA 的克隆、其在发育中的作用以及与细胞外基质糖蛋白的关系。

DOI：
10.1038/ki.1995.460
发表时间：
1995
期刊：
Kidney international
影响因子：
19.6
作者：
Kanwar,YS;Liu,ZZ;Kumar,A;Wada,J;Carone,FA
通讯作者：
Carone,FA

Identification of a renal-specific oxido-reductase in newborn diabetic mice.

DOI：
10.1073/pnas.160266197
发表时间：
2000-08
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Qiwei Yang;Bharat L. Dixit;Jun Wada;Yufeng Tian;E. Wallner;Satish K. Srivastva;Yashpal S Kanwar
通讯作者：
Qiwei Yang;Bharat L. Dixit;Jun Wada;Yufeng Tian;E. Wallner;Satish K. Srivastva;Yashpal S Kanwar

Ultrastructural changes of extracellular matrices in diabetic nephropathy revealed by high resolution scanning and immunoelectron microscopy.