TGF-beta superfamily in neonatal obstructive nephropathy and recovery

TGF-β超家族在新生儿梗阻性肾病及康复中的作用

• 批准号: 7633522
• 负责人: ROBERT L. CHEVALIER
• 金额: $ 34.65万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-17 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7633522
• 关键词: Adult; Animal Model; Animals; Apoptosis; Architecture; Binding; Binding Proteins; Biological Markers; Birth; Cell Culture Techniques; Cell Death; Cell Survival; Cell membrane; Cells; Cellular biology; Chemotaxis; Child; Chronic; Classification; Clinical Trials; Collagen; Complex; Congenital Abnormality; Contralateral; Cytoskeleton; Deposition; Development; Disease; Epithelial; Epithelial Cells; Event; Experimental Designs; Extracellular Matrix; Fetus; Fibroblasts; Fibrosis; Figs - dietary; Financial compensation; Genes; Genetic Transcription; Hour; Human; Immunohistochemistry; Impairment; In Vitro; Infant; Infiltration; Inflammation; Injury; Investigation; Kidney; Kidney Diseases; Kidney Failure; Knockout Mice; Lead; Lesion; Mediating; Membrane; Mesenchymal; Modeling; Molecular; Morphogenesis; Mus; Natural History; Neonatal; Newborn Infant; Nuclear; Obstruction; Operative Surgical Procedures; Pathologic; Pathway interactions; Peptides; Play; Proteins; Rattus; Receptor Activation; Recombinants; Recovery; Regulatory Element; Regulatory Pathway; Research; Role; Second Pregnancy Trimester; Severities; Signal Transduction; Signaling Molecule; Solid; Staging; Stretching; System; TGF-beta type I receptor; Testing; Therapeutic; Therapeutic Intervention; Time; Transforming Growth Factor beta; Transforming Growth Factors; Tubular formation; Ureteral obstruction; Urinary tract; Urine; Wild Type Mouse; Wound Healing; bone morphogenetic protein 7; bone morphogenetic protein receptor type II; cell injury; chordin; decorin; extracellular; follow-up; genetic manipulation; in vitro Model; indexing; inhibitor/antagonist; interstitial; macrophage; mature animal; member; metaplastic cell transformation; mouse model; mutant; partial recovery; postnatal; prevent; public health relevance; receptor; research study; response; transcription factor; urinary tract obstruction

DESCRIPTION (provided by applicant): Congenital obstructive nephropathy is the most important identifiable cause of renal impairment in infants and children. Despite this, indications for surgical intervention are controversial, and renal recovery is often disappointing. While mechanisms responsible for renal injury resulting from urinary tract obstruction are elucidated in models of complete unilateral ureteral obstruction (UUO) in adult animals, the developing kidney responds very differently to obstruction. The transforming growth factor-2 (TGF-2) superfamily plays a central role in renal development, wound healing, cell survival, phenotypic epithelial-mesenchymal transition (EMT), and fibrosis. This research plan will utilize a newly developed murine model to study the renal cellular response of the TGF-2 superfamily in regulating injury and recovery from partial UUO in the neonatal mouse, whose renal development is comparable to that of a 20-week human fetus. The model permits study of injury and recovery by the obstructed kidney, and compensation by the contralateral kidney. A combination of mutants and inhibitors of critical components of TGF-21 signaling, as well as of counter-regulatory bone morphogenetic protein-7 (BMP- 7) will be used to elucidate molecular mechanisms. Extracellular TGF-21 will be decreased by administration of decorin; TGF-21 receptor activity will be investigated by selective inhibition of ALK5; while TGF-21 intracellular signaling will be studied in Smad3 null mice. Extracellular BMP-7 will be manipulated in 3 ways: reduced endogenous BMP-7 activity in the kielin/chordin-like protein (KCP) null mouse; increased endogenous BMP-7 in the uterine sensitization-associated gene-1 (USAG-1) null mouse; and exogenous recombinant BMP-7. Indices of renal maturation, proliferation, apoptosis, and EMT, as well as macrophages, fibroblasts, and collagen deposition will be studied by immunohistochemistry and quantitative morphometrics. In vitro cell culture studies will be performed in parallel, and information gained (stretch-induced modulation of intracellular or junctional proteins) will be applied to the animal studies. The combination of surgical relief of obstruction with manipulation of TGF-21 may lead to new biomarkers or therapies to test in children with obstructive nephropathy. PUBLIC HEALTH RELEVANCE: Half of all children with kidney failure are born with birth defects of the urinary tract, and the most common of these are due to obstruction to urine flow. The timing and indications for surgical intervention are unclear, and recovery is unpredictable. A new animal (mouse) model is proposed to study the major cellular mechanisms for injury to the newborn kidney, as well as to evaluate new therapies to enhance recovery.

描述(申请人提供)：先天性梗阻性肾病是导致婴儿和儿童肾脏损害的最重要的可识别原因。尽管如此，手术干预的适应症仍存在争议，肾脏恢复也往往令人失望。尽管在成年动物单侧输尿管完全性梗阻(UUO)模型中阐明了尿路梗阻导致肾损伤的机制，但发育中的肾脏对梗阻的反应截然不同。转化生长因子-2超家族在肾脏发育、创伤愈合、细胞存活、表型上皮-间充质转化(EMT)和纤维化等过程中发挥重要作用。这项研究计划将利用新开发的小鼠模型来研究转化生长因子-2超家族在调节新生小鼠部分UUO的损伤和恢复中的肾脏细胞反应，其肾脏发育与20周的人类胎儿相当。该模型允许研究梗阻肾的损伤和恢复，以及对侧肾的代偿。转化生长因子-21信号关键成分的突变体和抑制剂以及骨形态发生蛋白-7(BMP-7)的逆调控物的组合将被用于阐明分子机制。在Smad3基因缺失的小鼠中，细胞外的转化生长因子-21将被降低；转化生长因子-21受体的活性将通过选择性抑制Alk5来检测；而转化生长因子-21的细胞内信号转导将被研究。细胞外BMP-7将通过三种方式进行操作：在Kielin/Chordin-like Protein(KCP)缺失小鼠中降低内源性BMP-7活性；在子宫致敏相关基因-1(USAG-1)缺失小鼠中增加内源性BMP-7活性；以及外源性重组BMP-7。通过免疫组织化学和定量形态计量学研究肾脏成熟、增殖、细胞凋亡和EMT的指标，以及巨噬细胞、成纤维细胞和胶原沉积。体外细胞培养研究将并行进行，所获得的信息(拉伸诱导的细胞内或连接蛋白的调节)将应用于动物研究。结合手术解除梗阻和操作转化生长因子-21可能会导致新的生物标记物或治疗方法用于梗阻性肾病儿童的检测。公共卫生相关性：所有肾衰竭儿童中有一半出生时就有尿路先天缺陷，其中最常见的是尿流受阻。手术干预的时机和适应症尚不清楚，恢复也是不可预测的。提出了一种新的动物(小鼠)模型，以研究新生儿肾脏损伤的主要细胞机制，以及评估促进恢复的新疗法。