AIM To delineate changes in miRNA expression localized to the peri-cystic local microenvironment(PLM) in an orthologous mouse model of autosomal dominant polycystic kidney disease(ADPKD)(mcwPkd1^(nl/nl)).METHODS We pr...AIM To delineate changes in miRNA expression localized to the peri-cystic local microenvironment(PLM) in an orthologous mouse model of autosomal dominant polycystic kidney disease(ADPKD)(mcwPkd1^(nl/nl)).METHODS We profiled miRNA expression in the whole kidney and laser captured microdissection(LCM) samples from PLM in mcwPkd1^(nl/nl)kidneys with Qiagen miScript 384 HC miRNA PCR arrays. The three times points used are:(1) post-natal(PN) day 21, before the development of trichrome-positive areas;(2) PN28, the earliest sign of trichrome staining; and(3) PN42 following the development of progressive fibrosis. PN21 served as appropriate controls and as the reference time point for comparison of miRNA expression profiles.RESULTS LCM samples revealed three temporally upregulated miRNAs [2 to 2.75-fold at PN28 and 2.5 to 4-fold(P ≤ 0.05) at PN42] and four temporally downregulated miRNAs [2 to 2.75 fold at PN28 and 2.75 to 5-fold(P ≤ 0.05) at PN42]. Expression of twenty-six miRNAs showed no change until PN42 [six decreased(2.25 to 3.5-fold)(P ≤ 0.05) and 20 increased(2 to 4-fold)(P ≤ 0.05)]. Many critical miRNA changes seen in the LCM samples from PLM were not seen in the contralateral whole kidney.CONCLUSION Precise sampling with LCM identifies miRNA changes that occur with the initiation and progression of renal interstitial fibrosis(RIF). Identification of the target proteins regulated by these miRNAs will provide new insight into the process of fibrosis and identify unique therapeutic targets to prevent or slow the development and progression of RIF in ADPKD.展开更多
基金Supported by the Children’s Research Institute,the Lillian Goldman Charitable TrustAmy P Goldman FoundationEllsworth Family and Children’s Foundation of Children’s’Hospital and Health System of Wisconsin
文摘AIM To delineate changes in miRNA expression localized to the peri-cystic local microenvironment(PLM) in an orthologous mouse model of autosomal dominant polycystic kidney disease(ADPKD)(mcwPkd1^(nl/nl)).METHODS We profiled miRNA expression in the whole kidney and laser captured microdissection(LCM) samples from PLM in mcwPkd1^(nl/nl)kidneys with Qiagen miScript 384 HC miRNA PCR arrays. The three times points used are:(1) post-natal(PN) day 21, before the development of trichrome-positive areas;(2) PN28, the earliest sign of trichrome staining; and(3) PN42 following the development of progressive fibrosis. PN21 served as appropriate controls and as the reference time point for comparison of miRNA expression profiles.RESULTS LCM samples revealed three temporally upregulated miRNAs [2 to 2.75-fold at PN28 and 2.5 to 4-fold(P ≤ 0.05) at PN42] and four temporally downregulated miRNAs [2 to 2.75 fold at PN28 and 2.75 to 5-fold(P ≤ 0.05) at PN42]. Expression of twenty-six miRNAs showed no change until PN42 [six decreased(2.25 to 3.5-fold)(P ≤ 0.05) and 20 increased(2 to 4-fold)(P ≤ 0.05)]. Many critical miRNA changes seen in the LCM samples from PLM were not seen in the contralateral whole kidney.CONCLUSION Precise sampling with LCM identifies miRNA changes that occur with the initiation and progression of renal interstitial fibrosis(RIF). Identification of the target proteins regulated by these miRNAs will provide new insight into the process of fibrosis and identify unique therapeutic targets to prevent or slow the development and progression of RIF in ADPKD.
