RATIONALE
A strong risk factor for atherosclerosis– the leading cause
of heart attacks and strokes– is the elevation of low-density
lipoprotein cholesterol (LDL-C) in blood. The LDL receptor (LDLR) is the
primary pathway for LDL-C removal from circulation, and their levels are
increased by statins --the main treatment for high blood LDL-C. However,
statins have low efficiency because they also increase PCSK9 which targets
LDLR for degradation. Since microRNAs have recently emerged as key
regulators of cholesterol homeostasis, our aim was to identify potential
microRNA-based therapeutics to decrease blood LDL-C and prevent
atherosclerosis.
METHODS AND RESULTS
We over expressed and knocked down miR-27a in HepG2 cells to assess
its effect on the expression of key players in the LDLR pathway using PCR
Arrays, Elisas, and Western blots. We found that miR-27a decreases LDLR
levels by 40% not only through a direct binding to its 3′
untranslated region but also indirectly by inducing a 3-fold increase in
PCSK9, which enhances LDLR degradation. Interestingly, miR-27a also directly
decreases LRP6 and LDLRAP1, two other key players in the LDLR pathway that
are required for efficient endocytosis of the LDLR-LDL-C complex in the
liver. The inhibition of miR-27a using lock nucleic acids induced a
70% increase in LDLR levels and, therefore, it would be a more
efficient treatment for hypercholesterolemia because of its desirable
effects not only on LDLR but also on PCSK9.
CONCLUSION
The results presented here provide evidence supporting the potential
of miR-27a as a novel therapeutic target for the prevention of
atherosclerosis.
Individuals with type 2 diabetes have an increased risk of developing nonalcoholic fatty liver disease (NAFLD), and NAFLD patients are also at greater risk for developing type 2 diabetes. Although the relationship between type 2 diabetes and NAFLD is highly interconnected, the pathogenic mechanisms linking the two diseases are poorly understood. The goal of this study was to identify genetic determinants of hepatic lipid accumulation through association analysis using histological phenotypes in obese individuals. Using the Illumina HumanOmniExpress BeadChip assay, we genotyped 2300 individuals on whom liver biopsy data were available. We analyzed total bilirubin levels, which are linked to fatty liver in severe obesity, and observed the strongest evidence for association with rs4148325 in UGT1A (P<5.0 × 10−93), replicating previous findings. We assessed hepatic fat level and found strong evidence for association with rs4823173, rs2896019, and rs2281135, all located in PNPLA3 and rs10401969 in SUGP1. Analysis of liver transcript levels of 20 genes residing at the SUGP1/NCAN locus identified a 1.6-fold change in expression of the LPAR2 gene in fatty liver. We also observed suggestive evidence for association between low-grade fat accumulation and rs10859525 and rs1294908, located upstream from SOCS2 and RAMP3, respectively. SOCS2 was differentially expressed between fatty and normal liver. These results replicate findings for several hepatic phenotypes in the setting of extreme obesity and implicate new loci that may play a role in the pathophysiology of hepatic lipid accumulation.
Nephrin, a major component of the glomerular slit diaphragm (SD), is both a structural protein as well as a signaling molecule influencing foot process (FP) formation and maintenance of podocyte integrity. Analyses of near-term embryonic kidneys showed normal cellular viability and no apoptosis in glomeruli from nephrin knockout mice. Moreover, expression and location of other SD or glomerular basement membrane components were similar in wild-type and mutant mice as was the location and levels of most podocyte-specific proteins. Transcriptional profiling showed that the lack of nephrin had minor impact on the expression of genes for FPs and SD proteins. Claudin 3, a tight-junction protein normally absent in glomeruli, was upregulated threefold in the knockout mice, suggesting a role of nephrin in claudin 3 gene expression within the glomeruli. Our results suggest that nephrin is expressed late in the process of podocyte differentiation and is a locus for the formation of SD and FP maintenance and physical integrity in vivo. Nephrin does not seem to have a primary role in cell survival but has a small impact on gene regulation during glomerular development.
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