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2014 ; 86
(6
): 1116-29
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Discovery of new glomerular disease-relevant genes by translational profiling of
podocytes in vivo
#MMPMID24940801
Grgic I
; Hofmeister AF
; Genovese G
; Bernhardy AJ
; Sun H
; Maarouf OH
; Bijol V
; Pollak MR
; Humphreys BD
Kidney Int
2014[Dec]; 86
(6
): 1116-29
PMID24940801
show ga
Identifying new biomarkers and therapeutic targets for podocytopathies such as
focal segmental glomerulosclerosis (FSGS) requires a detailed analysis of
transcriptional changes in podocytes over the course of disease. Here we used
translating ribosome affinity purification (TRAP) to isolate and profile
podocyte-specific mRNA in two different models of FSGS. We expressed enhanced
green fluorescent protein-tagged to ribosomal protein L10a in podocytes under the
control of the collagen-1?1 promoter, enabling one-step podocyte-specific mRNA
isolation over the course of disease. This TRAP protocol robustly enriched known
podocyte-specific mRNAs. We crossed Col1?1-eGFP-L10a mice with the Actn4(-/-) and
Actn4(+/K256E) models of FSGS and analyzed podocyte transcriptional profiles at
2, 6, and 44 weeks of age. Two upregulated podocyte genes in murine FSGS (CXCL1
and DMPK) were found to be upregulated at the protein level in biopsies from
patients with FSGS, validating this approach. There was no dilution of
podocyte-specific transcripts during disease. These are the first
podocyte-specific RNA expression data sets during aging and in two models of
FSGS. This approach identified new podocyte proteins that are upregulated in FSGS
and defines novel biomarkers and therapeutic targets for human glomerular
disease.