Laser capture microdissection and multiplex-tandem PCR analysis of proximal tubular epithelial cell signaling in human kidney disease

Wilkinson, Ray, Wang, Xiangju, Kassianos, Andrew J., Zuryn, Steven, Roper, Kathrein E., Osborne, Andrew, Sampangi, Sandeep, Francis, Leo, Raghunath, Vishwas and Healy, Helen (2014) Laser capture microdissection and multiplex-tandem PCR analysis of proximal tubular epithelial cell signaling in human kidney disease. PLoS One, 9 1: e87345.1-e87345.10. doi:10.1371/journal.pone.0087345


Author Wilkinson, Ray
Wang, Xiangju
Kassianos, Andrew J.
Zuryn, Steven
Roper, Kathrein E.
Osborne, Andrew
Sampangi, Sandeep
Francis, Leo
Raghunath, Vishwas
Healy, Helen
Title Laser capture microdissection and multiplex-tandem PCR analysis of proximal tubular epithelial cell signaling in human kidney disease
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2014-01
Sub-type Article (original research)
DOI 10.1371/journal.pone.0087345
Open Access Status DOI
Volume 9
Issue 1
Start page e87345.1
End page e87345.10
Total pages 10
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Collection year 2015
Language eng
Abstract Interstitial fibrosis, a histological process common to many kidney diseases, is the precursor state to end stage kidney disease, a devastating and costly outcome for the patient and the health system. Fibrosis is historically associated with chronic kidney disease (CKD) but emerging evidence is now linking many forms of acute kidney disease (AKD) with the development of CKD. Indeed, we and others have observed at least some degree of fibrosis in up to 50% of clinically defined cases of AKD. Epithelial cells of the proximal tubule (PTEC) are central in the development of kidney interstitial fibrosis. We combine the novel techniques of laser capture microdissection and multiplex-tandem PCR to identify and quantitate “real time” gene transcription profiles of purified PTEC isolated from human kidney biopsies that describe signaling pathways associated with this pathological fibrotic process. Our results: (i) confirm previous in-vitro and animal model studies; kidney injury molecule-1 is up-regulated in patients with acute tubular injury, inflammation, neutrophil infiltration and a range of chronic disease diagnoses, (ii) provide data to inform treatment; complement component 3 expression correlates with inflammation and acute tubular injury, (iii) identify potential new biomarkers; proline 4-hydroxylase transcription is down-regulated and vimentin is up-regulated across kidney diseases, (iv) describe previously unrecognized feedback mechanisms within PTEC; Smad-3 is down-regulated in many kidney diseases suggesting a possible negative feedback loop for TGF-β in the disease state, whilst tight junction protein-1 is up-regulated in many kidney diseases, suggesting feedback interactions with vimentin expression. These data demonstrate that the combined techniques of laser capture microdissection and multiplex-tandem PCR have the power to study molecular signaling within single cell populations derived from clinically sourced tissue.
Keyword Gene-expression profiles
Injury molecule-1 Kim-1
Mesenchymal transition
Diabetic-nephropathy
Renal-disease
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
School of Medicine Publications
 
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