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Kidney Int DOI:10.1038/ki.2014.204

Discovery of new glomerular disease-relevant genes by translational profiling of podocytes in vivo.

Publication TypeJournal Article
Year of Publication2014
AuthorsGrgic, I, Hofmeister, AF, Genovese, G, Bernhardy, AJ, Sun, H, Maarouf, OH, Bijol, V, Pollak, MR, Humphreys, BD
JournalKidney Int
Volume86
Issue6
Pages1116-29
Date Published2014 Dec
ISSN1523-1755
KeywordsActinin, Aging, Animals, Biomarkers, Chemokine CXCL1, Collagen Type I, Disease Models, Animal, Gene Expression Profiling, Glomerulosclerosis, Focal Segmental, Green Fluorescent Proteins, Humans, Mice, Mice, Knockout, Myotonin-Protein Kinase, Neoplasm Proteins, Oligonucleotide Array Sequence Analysis, Podocytes, Protein Biosynthesis, Ribosomal Proteins, RNA, Messenger, Transcriptome
Abstract

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-1α1 promoter, enabling one-step podocyte-specific mRNA isolation over the course of disease. This TRAP protocol robustly enriched known podocyte-specific mRNAs. We crossed Col1α1-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.

URLhttp://dx.doi.org/10.1038/ki.2014.204
DOI10.1038/ki.2014.204
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/24940801?dopt=Abstract

Alternate JournalKidney Int.
PubMed ID24940801
PubMed Central IDPMC4245460
Grant ListR01 DK088923 / DK / NIDDK NIH HHS / United States
R01 DK054931 / DK / NIDDK NIH HHS / United States
DK54931 / DK / NIDDK NIH HHS / United States
R56 DK054931 / DK / NIDDK NIH HHS / United States
R37 DK059588 / DK / NIDDK NIH HHS / United States
DK087389 / DK / NIDDK NIH HHS / United States
DK88923 / DK / NIDDK NIH HHS / United States
RC1 DK087389 / DK / NIDDK NIH HHS / United States
P30 DK081943 / DK / NIDDK NIH HHS / United States
T32 DK007527 / DK / NIDDK NIH HHS / United States