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，因为高血糖会对胚胎和成年期间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，例如泌尿生殖系统发育不全/发育不全，以及成人糖尿病肾病。在糖尿病肾病中，肾小球和肾小管间质室均受累，观察到细胞外基质（ECM）增加。 ECM 积累是由于基质蛋白（例如胶原蛋白和纤连蛋白）合成增加所致 MMP 活性降低，ECM 蛋白糖化，形成高级糖化产物 (AGE)，导致活性氧 (ROS) 的产生，活性氧通过下游信号，如 TGF-β，诱导进行性肾小球和肾小管-间质损伤。此外，还报道了其他 ECM 蛋白（例如骨桥蛋白和纤连蛋白）表达的改变。然而，最近鉴定的 ECM 蛋白（肾小管间质蛋白）的作用糖尿病肾病中的肾炎抗原（TIN-ag）仍然是个谜。 TIN-ag 是一种镶嵌蛋白，含有组织蛋白酶 B 样结构域、卵泡抑素模块、ATP 结合基序、糖基化位点以及与其他 ECM 蛋白的片段同源性。它受到发育调节，在肾小管祖细胞中表达，并在体外调节肾小管发生。为了破译 TIN-ag 在胚胎和成年期间进行性肾损伤中的作用，根据以下具体目标提出了实验： I. TIN-ag 在葡萄糖诱导的肾脏畸形发生中的作用将被研究自从用高葡萄糖或 TIN-ag 反义物处理胎儿小鼠肾脏后进行了研究，产生了相似的表型，即抑制肾小管发生，表明 TIN-ag 在葡萄糖诱导病变的演变中可能存在相关性。二.将使用Tet-On/Off系统在转基因小鼠和胎儿Lewis大鼠中研究TIN-ag在过度表达和掩蔽TIN-ag基因的动物模型中的作用。三． TIN-ag 及其与其他 ECM 蛋白的域特异性相互作用将由于 TIN-ag 在成年 Lewis 大鼠中被掩盖，因此进行了评估。将产生重组 TIN-ag 以研究蛋白质：蛋白质与 IV 型胶原和层粘连蛋白亚型的相互作用，并且酵母双杂交系统将鉴定新的相互作用伙伴。四． TIN-ag在多种糖尿病肾病动物模型中的作用及相关机制将通过形态学和生化研究来研究其表达增加。体外培养系统也将用于描述糖尿病环境中 TIN-ag 表达改变的机制。 V. 将通过生成 TIN-ag 条件敲除和常规敲除 (KO) 小鼠来研究 TIN-ag 在肾小管间质病理学中的作用。为了产生条件性 KO 小鼠，杂合子痛风小鼠将与 Ella Cre 杂交，然后与病变仅限于肾脏的 Ksp Cre 小鼠杂交。 Null 等位基因将进行杂交，产生 Null -/- 突变体，作为传统 KO 小鼠，并消除 TIN-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）

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

DOI：
发表时间：
1993
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
0
作者：
H. Makino;Y. Yamasaki;T. Haramoto;K. Shikata;K. Hironaka;Z. Ota;Y. Kanwar
通讯作者：
H. Makino;Y. Yamasaki;T. Haramoto;K. Shikata;K. Hironaka;Z. Ota;Y. Kanwar

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

Influence of genetics on the nephritogenic potential of proteoglycans.

遗传学对蛋白多糖肾炎发生潜力的影响。