The expression level of the liver low density lipoprotein receptor (LDLR) is one of the most important regulators of human plasma LDL cholesterol (LDL-c). Increased hepatic LDLR expression results in an improved clearance of LDL-c from circulation, hence directly reducing the risk of coronary heart disease. Currently, in the research area of LDLR, the transcriptional regulation of the LDLR gene has been extensively investigated and most of the pharmaceutical interventions are aimed to increase LDLR gene transcription, although indirectly, in order to lower plasma LDL-c. However, the posttranscriptional modulations of LDLR are much understudied and the mechanisms controlling LDLR mRNA turnover are largely unknown. By conducting a screening of 700 compounds isolated from Chinese herbs, we have identified Berberine (BBR), a compound originally isolated from the Chinese herb Huanglian, as a novel upregulator of hepatic LDLR. BBR strongly increased the LDLR mRNA and protein expression in human hepatoma-derived cells and it also effectively reduced the serum LDL-c in hypercholesterolemic patients when tested in a placebo-controlled clinical study. Further investigations reveal that BBR increases the LDLR expression by a unique posttranscriptional mechanism involving LDLR mRNA stabilization through its 3'-untranslated region (3'UTR). We have now further localized the BBR-responsive regulatory sequences to a 905 nt segment in the 5'proximal region of LDLR 3'UTR. The overall objectives of this new proposal are to fully elucidate the molecular and cellular mechanisms by which BBR regulates LDL mRNA stability in cell culture and in animal models.
The specific aims of this new proposal are to: 1) identify the BBRresponsive cis-regulatory elements residing in the LDLR mRNA 3'UTR through a series of mutational and functional analyses of a panel of reporter constructs that contain the wild-type and mutated cis-regulatory sequences of the 905 nt BBR-responsive region of the 3'UTR;2) characterize the mRNA-binding proteins that specifically interact with the BBR-responsive elements;and 3) demonstrate the in vivo activities of BBR in stabilizing the hepatic LDLR mRNA and the consequential LDL-c lowering in animal models. The successful accomplishment of these specific aims will have profound impacts to the current research area of hypercholesterolemia. A clear understanding at the molecular level of how LDLR mRNA stability is regulated in terms of cis-acting regulatory sequences and trans-acting factors will provide an important foundation for exploiting the new therapeutic approach that increases liver LDLR expression through mRNA stabilization.

Agency
National Institute of Health (NIH)
Institute
National Center for Complementary & Alternative Medicine (NCCAM)
Type
Research Project (R01)
Project #
5R01AT002543-05
Application #
7679389
Study Section
Hepatobiliary Pathophysiology Study Section (HBPP)
Program Officer
Hopp, Craig
Project Start
2005-09-30
Project End
2012-09-29
Budget Start
2009-09-30
Budget End
2012-09-29
Support Year
5
Fiscal Year
2009
Total Cost
$327,762
Indirect Cost
Name
Palo Alto Institute for Research & Edu, Inc.
Department
Type
DUNS #
624218814
City
Palo Alto
State
CA
Country
United States
Zip Code
94304
Kan, Chin Fung Kelvin; Singh, Amar Bahadur; Dong, Bin et al. (2015) PPAR? activation induces hepatic long-chain acyl-CoA synthetase 4 expression in vivo and in vitro. Biochim Biophys Acta 1851:577-87
Dong, Bin; Li, Hai; Singh, Amar Bahadur et al. (2015) Inhibition of PCSK9 transcription by berberine involves down-regulation of hepatic HNF1? protein expression through the ubiquitin-proteasome degradation pathway. J Biol Chem 290:4047-58
Kan, Chin Fung Kelvin; Singh, Amar Bahadur; Stafforini, Diana M et al. (2014) Arachidonic acid downregulates acyl-CoA synthetase 4 expression by promoting its ubiquitination and proteasomal degradation. J Lipid Res 55:1657-67
Singh, Amar Bahadur; Kan, Chin Fung Kelvin; Shende, Vikram et al. (2014) A novel posttranscriptional mechanism for dietary cholesterol-mediated suppression of liver LDL receptor expression. J Lipid Res 55:1397-407
Dong, Bin; Singh, Amar Bahadur; Fung, Chin et al. (2014) CETP inhibitors downregulate hepatic LDL receptor and PCSK9 expression in vitro and in vivo through a SREBP2 dependent mechanism. Atherosclerosis 235:449-62
Singh, Amar Bahadur; Li, Hai; Kan, Chin Fung Kelvin et al. (2014) The critical role of mRNA destabilizing protein heterogeneous nuclear ribonucleoprotein d in 3' untranslated region-mediated decay of low-density lipoprotein receptor mRNA in liver tissue. Arterioscler Thromb Vasc Biol 34:8-16
Dong, Bin; Kan, Chin Fung Kelvin; Singh, Amar B et al. (2013) High-fructose diet downregulates long-chain acyl-CoA synthetase 3 expression in liver of hamsters via impairing LXR/RXR signaling pathway. J Lipid Res 54:1241-54
Li, Hai; Liu, Jingwen (2012) The novel function of HINFP as a co-activator in sterol-regulated transcription of PCSK9 in HepG2 cells. Biochem J 443:757-68
Wu, Minhao; Dong, Bin; Cao, Aiqin et al. (2012) Delineation of molecular pathways that regulate hepatic PCSK9 and LDL receptor expression during fasting in normolipidemic hamsters. Atherosclerosis 224:401-10
Dong, Bin; Wu, Minhao; Cao, Aiqin et al. (2011) Suppression of Idol expression is an additional mechanism underlying statin-induced up-regulation of hepatic LDL receptor expression. Int J Mol Med 27:103-10

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