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. 2013 Aug 1;319(13):2073-2080.
doi: 10.1016/j.yexcr.2013.04.011. Epub 2013 Apr 21.

Rapamycin-induced Modulation of miRNA Expression Is Associated With Amelioration of HIV-associated Nephropathy (HIVAN)

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Free PMC article

Rapamycin-induced Modulation of miRNA Expression Is Associated With Amelioration of HIV-associated Nephropathy (HIVAN)

Kang Cheng et al. Exp Cell Res. .
Free PMC article

Abstract

Recent studies suggested that miRNAs are involved in the development of the pathogenesis of HIV-associated nephropathy (HIVAN). Rapamycin, a widely used mTOR inhibitor, has been demonstrated to slow down the progression of HIVAN. However, the role of miRNA in the regulation of these processes has not been investigated so far. In the current study, we have used a microarray-based approach in combination with real-time PCR to profile the miRNA expression patterns in rapamycin-treated HIVAN mice (Tg26). Our results demonstrated that 19 miRNAs belonging to 13 different families expressed differentially in renal tissues of rapamycin-receiving Tg26 mice when compared to Tg26 mice-receiving saline only. The patterns of miRNAs expression in rapamycin-receiving Tg26 mice took a reverse turn. These miRNAs were classified into 8 functional categories. In in vitro studies, we examined the expression of specific miRNAs in HIV-1 transduced human podocytes (HIV/HPs). HIV/HPs displayed attenuation of expression of miR-99a, -100a, -199a and miR-200, whereas, rapamycin inhibited this effect of HIV. These findings suggest that rapamycin-mediated up-regulation of specific miRNAs could contribute to amelioration of renal lesions in HIVAN mice.

Keywords: Apoptosis; Epithelial mesanchymal transition; HIV-associated nephropathy; Mammalian target of rapamycin; MicroRNA.

Figures

Fig. 1
Fig. 1
Rapamycin attenuates renal lesions in HIVAN mice. (A) Representative microphotographs of FVB/N, Tg26 and rapamycin-treated Tg26 mice. Tg26 mice-receiving normal saline showed sclerosed glomerulus and tubular cyst filled with proteinaceous cast. Rapamycin-treated Tg26 mice showed minimal tubular dilatation without glomerulosclerosis. (B) Cumulative data showing percentage of normal glomeruli, sclerosed glomeruli and collapsing glomeruli.
Fig. 2
Fig. 2
Differential expression of miRNAs in kidneys of Tg26 and rapamycin-treated Tg26 mice. (A) Heatmap depicts triplicate microarray hybridizations, revealing a subset of miRNAs that are differentially expressed in rapamycin-treated Tg26 mice compared with Tg26 and control FVB/N mice. The color scale shown at the bottom: green denotes expression >40 and red denotes an expression <0. (B) The relative expression of miRNAs level is present in log2 transformation. Total 3 miRNAs were upregulated (right) and 16 miRNAs were downregulated (left) in rapamycin-treated Tg26 mice (P<0.05).
Fig. 3
Fig. 3
Validation and confirmation of miRNAs expression by real-time quantitative PCR. qPCR demonstrated that rapamycin led to inverse expression patterns of miRNAs in Tg26 mice. Of these, miR-466i, miR-467f and miR-669a were up regulated and miR-497, miR-140, miR-145, miR-331, miR-16, miR-30a, miR-22, miR-30c, miR-378, miR-99a, miR-100a, miR-199a, miR-200a, miR-141, miR-200b, miR-200c and miR-429 were down regulated at 8 wks in HIVAN mice. U6 snRNA was used as an internal control. The relative expression level of specific miRNA was calculated using modified 2−ΔΔCt method.
Fig. 4
Fig. 4
Overrepresented functional categories of the differential expressed miRNAs in HIVAN. (A) The pie-chart shows 8 functional categories and each pie represents a functional category with an overrepresentation of regulatory pathways of miRNA targets. (B) Overlapping of miRNAs in Wnt pathway, oxidative pathway, and EMT pathway.
Fig. 5
Fig. 5
MiRNAs expression in HIV-transduced human podocytes (HIVHPs). HPs were infected with HIV-1 virus for 72 h. Real-time PCR analysis showed that HIV induced up regulation of miR-99a, miR-100a, miR-199a, miR-200a, miR-200b, miR-200c, miR-429 and miR-141; whereas rapamycin led to inverse expression of these miRNAs in HIV/HPs. U6 snRNA was used as an internal control. The relative expression level of specific miRNA was calculated using modified 2−ΔΔCt method.

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