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Motor protein Myo1c participates in Nephrin and Neph1 signaling

运动蛋白 Myo1c 参与 Nephrin 和 Neph1 信号传导

基本信息

批准号：
9094533
负责人：
DEEPAK NIHALANI
金额：
$ 49.01万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9094533
关键词：
Actins Acute Adriamycin PFS Affect Agreement Attenuated Binding Biology Breeding Cell membrane Cells Chronic Clathrin Collaborations Communities Complex Cytoskeleton Development Diabetic Nephropathy Disease Emotional End stage renal failure Endocytosis Endosomes Event Goals Hair Cells Health Human Impairment Injury Intracellular Membranes Kidney Failure Knock-out Knockout Mice Lead Life Link LoxP-flanked allele Lysosomes Measures Mediating Membrane Modeling Molecular Motor Movement Mus Nephrology Phosphorylation Play Positioning Attribute Protamine Sulfate Proteins Proteinuria Recycling Renal function Renal glomerular disease Reporting Resistance Role Signal Transduction Staging Stimulus Sushi Domain System Testing Therapeutic Vesicle Zebrafish aged base coated pit disease phenotype effective therapy efficacy testing extracellular glomerular filtration glomerular function hearing impairment in vivo inhibitor/antagonist late endosome mouse development mouse model nephrin novel podocyte prevent response response to injury slit diaphragm therapeutic target trafficking trans-Golgi Network

项目摘要

DESCRIPTION (provided by applicant): We identified motor protein Myo1c as a novel component of slit diaphragm that interacts with Nephrin and Neph1 and regulates their movement. To study the in vivo function of Myo1c, we generated Myo1c floxed mice that were used to construct homozygous null and podocyte-specific Myo1c null mice. While Myo1c homozygous null mice were reported to die prenatally, complete Myo1c deletion in 12 week old mice increased their sensitivity to adriamycin-induced proteinuria (on a C57BL/6J background), which is in agreement with our zebrafish studies. Interestingly, the analysis of podocyte specific Myo1c knockout mouse showed no proteinuria or functional abnormality when aged to 8 months. However, when bred to an adriamycin-sensitive background, these mice were resistant to adriamycin-induced glomerulopathy; they did not develop proteinuria as compared to the control mice suggesting multiple functions for Myo1c. Although surprising, these results are consistent with loss of proteins in podocytes that are involved in signaling and trafficking such as Rac1 and Crk. Podocytes response to injury is commonly assessed through increased phosphorylation of Nephrin and Neph1 that initiates their redistribution and the assembly of an intracellular signaling cascade leading to podocyte effacement. Impairment of these events may attenuate podocytes ability to respond to injury thus inducing protection. Indeed, our recent study demonstrated that inhibiting Neph1 signaling protected podocytes from injury. Since Myo1c has a tethering function that associates its cargo proteins with membranes and actin, we hypothesized that Myo1c participates in a mechanism that regulates movement of these proteins at the membrane that is critical for directing the assembly of a signaling complex by Nephrin and Neph1 and initiating intracellular signaling and trafficking events. Indeed, loss of Myo1c binding attenuated the dynamic movement of Neph1 at the membrane as demonstrated using live FRAP analysis. Furthermore, we treated cultured podocytes with a Myo1c specific inhibitor pentachloropseudilin (PCIP) that arrested membrane and intracellular vesicles movements suggesting the involvement of Myo1c in actin dependent cellular events that are critical for generating cellular response to injury. This further suggested that these cells will hve impaired injury response. Indeed, these cells resisted injury by protamine sulphate (PS) as measured by actin cytoskeleton reorganization. In the Specific Aim 1, we will investigate the hypothesis that Myo1c due to its membrane and actin binding functions, participates in generating an appropriate injury response by podocytes. This involves regulating injury-induced redistribution of Nephrin and Neph1 proteins to intracellular compartments and the assembly of signaling complexes that drives their intracellular signaling and trafficking. In the second Aim, we will investigate how Myo1c depletion at various stages of mouse development affects glomerular function. In addition, we will investigate if Myo1c is a therapeutic target by determining whether podocyte-specific deletion of Myo1c attenuates the disease phenotype in various acute and chronic glomerular injury models.