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Series GSE139585 Query DataSets for GSE139585
Status Public on Aug 23, 2020
Title A Uromodulin mutation resulting in kidney fibrosis due to activation of the innate immune system through mononuclear phagocyte ATF4/integrated stress response signaling
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Summary Background: Uromodulin (Tamm-Horsfall) protein is secreted by the thick ascending limb into urine and crosses the basolateral membrane into the kidney interstitium and blood. Uromodulin may engage the kidney’s innate immune system with pro and anti-inflammatory effects described. Mutations in the UMOD gene are a cause of autosomal dominant tubulointerstitial kidney disease (ADTKD) due to protein misfolding and endoplasmic reticulum stress. We identified a family with a UMOD mutation (C106F) that leads to glomerular and interstitial inflammation atypical for ADTKD.
Methods: To determine if the observed phenotype is due to mutant uromodulin induced changes in the innate or adaptive immune system, we developed a transgenic (tg) mouse with a homologous cysteine to phenylalanine mutation (C105F) in the UMOD gene using CRISPR-Cas9 gene editing.
Results: LLC-PK1 cells expressing wt and mutant protein had increased basolateral secretion of protein compared with cells expressing wt or mutant protein alone. EM examination of mutant medium showed protein aggregates in contrast to filaments in wt medium. Immunoprecipitation of plasma uromodulin from mutant mice demonstrated aggregate formation not seen in wt mice. Tg/+ mice displayed increased F4/80+ mononuclear phagocyte (MP) and podocyte inflammasome activity at baseline. Tg/+ mice developed glomerular and interstitial matrix deposition, myofibroblast proliferation and increased Creatinine with aging. Following ischemia-reperfusion injury, tg/+ mice had no increase in inflammasome activation over baseline in contrast to wt littermates and displayed improved tubular repair and renal function at 7 days. Differential gene expression analysis by RNA sequencing of kidney MP revealed activation of the ATF4 mediated stress response pathway and genes associated with the development of tissue fibrosis.
Conclusions: The C106F mutation results in glomerular and interstitial inflammatory kidney disease due to interaction of aggregated misfolded protein with mononuclear phagocytic cells resulting in ER stress response. The resulting alteration in the immune system leads to improved kidney repair following acute injury but fibrosis and chronic renal failure with aging.
Overall design To determine if changes in macrophage gene expression is a mechanism linking phagocytosis of aggregated uromodulin to the pathogenesis of disease in transgenic mice, we performed RNA-sequencing of mRNA isolated from F4/80+ cells purified from 4 mutant mice and 6 wt littermate controls at 12-16 weeks using anti-F4/80 conjugated magnetic microbeads. Total N=10.
Contributor(s) Plotkin M, Obrien C, Goellner J, Williams J, Carter W, Stone A
Citation(s) 32926855
Submission date Oct 30, 2019
Last update date Sep 17, 2020
Contact name Annjanette Stone
Phone 501-257-5178
Organization name CAVHS
Department Research
Lab Pharmacogenomics Analysis Laboratory
Street address 4300 West 7th Street
City Little Rock
State/province AR
ZIP/Postal code 72205
Country USA
Platforms (1)
GPL24247 Illumina NovaSeq 6000 (Mus musculus)
Samples (10)
GSM4143989 TG1
GSM4143990 TG2
GSM4143991 TG4
BioProject PRJNA580376
SRA SRP227445

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SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE139585_Expression_Profile.mm10.transcript.xlsx 5.6 Mb (ftp)(http) XLSX
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Raw data are available in SRA
Processed data are available on Series record

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