Inducible Knockout of Par 1 a/b in the Kidney

在肾脏中诱导击倒标准杆 1 a/b

• 批准号: 9135837
• 负责人: Kimberly Jean Reidy
• 金额: $ 8.32万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9135837
• 关键词: Acute Kidney Tubular Necrosis; Acute Renal Failure with Renal Papillary Necrosis; Adult; Affect; Alleles; Area; Biological Markers; Biopsy; Birth; Breeding; Cadherins; Cell Culture Techniques; Cell Cycle; Cell Survival; Cell-Cell Adhesion; Cisplatin; Clustered Regularly Interspaced Short Palindromic Repeats; Creatinine; Data; Databases; Development; Diagnosis; Diet; Dose; Doxycycline; Embryo; Epithelial; Excision; Exons; Future; Generations; Genome engineering; Health Care Costs; Hematoxylin and Eosin Staining Method; Histology; Homologous Gene; Hour; Human; Individual; Injection of therapeutic agent; Injury; K-Series Research Career Programs; Kidney; Knock-out; Knockout Mice; Lead; Left; Mediating; Modeling; Morbidity - disease rate; Mus; Mutant Strains Mice; Organ; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Protein-Serine-Threonine Kinases; Proteins; Recovery; Renal function; Resected; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Severities; Signal Pathway; Signal Transduction; Site; Source; Specimen; Staining method; Stains; Testing; Tetanus Helper Peptide; Therapeutic; Tissue Model; Tissues; Transgenic Mice; Transgenic Organisms; Transplantation; Tubular formation; Western Blotting; beta catenin; clinical Diagnosis; embryonic stem cell; feeding; human disease; improved; intraperitoneal; light microscopy; mortality; mouse model; mutant; nephrogenesis; notch protein; pregnant; public health relevance; renal epithelium; repaired; tool; translational study; tumor

 DESCRIPTION (provided by applicant): This proposal involves development of a new mouse model and generation of preliminary data that will facilitate transition from the current K award t independence. Acute kidney injury (AKI) is associated with morbidity and mortality in hospitalized patients. Apico-basal polarity is required for directional transport in renal epitheli and is disrupted in AKI. Partitioning defective Par1a and 1b are serine threonine kinases that establish apico-basal polarity in cell culture, but also regulate downstream pathways that affect cell-cell adhesion, cell cycle, cell survival and other pathways that are relevant for epithelial repair. The underlying hypothesis of these studies is that Par1a/b are protective in the setting of acute kidney injury and contribute to renal epithelial repair. To test this, we will generate a mouse model for inducible kidney specific deletion of Par1a/b using CRISPR/Cas9 mediated genome engineering, and examine the effect of loss of Par1a/b on severity of acute kidney injury. In addition, we will examine the relevance of Par1a/b induction after injury, by examining expression of Par1a/b in human kidney tissue (left over, not needed for diagnosis) with pathologic evidence of AKI. The specific aims are: Aim1: Generate a model of inducible kidney specific deletion of Par1a/b. Aim2: Determine effect of Par1a/b deletion on severity of acute kidney injury in mice, and Aim3: Examine the expression of Par1a/b in human acute kidney injury.

 描述（由申请人提供）：该提案涉及开发一种新的小鼠模型和生成初步数据，这将有助于从目前的K奖过渡到独立性。急性肾损伤（阿基）与住院患者的发病率和死亡率相关。肾上皮细胞中的定向转运需要顶端-基底极性，并且在阿基中被破坏。分配缺陷型Par 1a和1b是丝氨酸苏氨酸激酶，其在细胞培养中建立顶-基底极性，但也调节影响细胞-细胞粘附、细胞周期、细胞存活的下游途径和与上皮修复相关的其他途径。这些研究的基本假设是Par 1a/B在以下情况下具有保护作用： 急性肾损伤，并有助于肾上皮修复。为了测试这一点，我们将使用CRISPR/Cas9介导的基因组工程生成用于Parla/B的可诱导肾特异性缺失的小鼠模型，并检查Parla/B的缺失对急性肾损伤严重程度的影响。此外，我们将通过检查具有阿基病理学证据的人肾组织（剩余，不需要诊断）中的Par 1a/B表达，来检查损伤后Par 1a/B诱导的相关性。具体目标是：目标1：生成可诱导的Par 1a/B肾特异性缺失模型。目标2：确定Par 1a/B缺失对小鼠急性肾损伤严重程度的影响，以及目的3：检测人急性肾损伤中Par 1a/B的表达。