Preparation and activity analysis of recombinant human high-density lipoprotein
Manman Su1, Weiqin Chang, Kaiyao Shi
1Department of Regenerative Medicine, College of Pharmacy, Jilin University, Changchun, People's Republic of China.
Insights
Recombinant human high-density lipoprotein (rhHDL) mimics native HDL's function. This new therapeutic agent effectively removes cholesterol and combats atherosclerosis.
Area of Science:
- Biochemistry
- Cardiovascular Research
- Biotechnology
Background:
- Plasma high-density lipoprotein (HDL) levels inversely correlate with atherosclerotic cardiovascular disease risk.
- HDL is a key therapeutic target, but native HDL has application limitations.
Purpose of the Study:
- To develop a recombinant human HDL (rhHDL) with native HDL's composition and function.
- To evaluate rhHDL's therapeutic potential as an HDL-replacement agent.
Main Methods:
- Expressed and purified six recombinant human apolipoproteins (rhapo) in Pichia pastoris.
- Assembled rhapo with phosphatidylcholine and cholesterol into rhHDL using cholate and density-gradient centrifugation.
- Conducted in vitro and in vivo comparative studies between native HDL and rhHDL.
Main Results:
- rhHDL successfully formed micellar complexes with lipids.
- In vitro assays demonstrated rhHDL promotes cholesterol efflux from macrophages.
- In vivo studies showed rhHDL effectively modulates blood lipid metabolism in hyperlipidemic mice, similar to native HDL.
Conclusions:
- rhHDL exhibits anti-atherosclerotic properties comparable to native HDL.
- These properties include promoting reverse cholesterol transport, antioxidative, and antithrombotic effects.
- rhHDL shows promise as a therapeutic agent for cardiovascular diseases.
Abstract:
Population studies have consistently shown a highly inverse correlation between plasma concentration of high-density lipoprotein and the risk of atherosclerotic cardiovascular disease in humans. High-density lipoprotein (HDL) as a therapeutic target is an intense area of ongoing investigation. Aiming to solve the shortcomings of native HDL application, we prepared recombinant human HDL (rhHDL) that contains a similar composition and has similar functions with native HDL. Six kinds of recombinant human apolipoproteins (rhapo)-rhapoA-I, rhapoA-II, rhapoA-IV, rhapoC-I, rhapoC-II, and rhapoE-were expressed in Pichia pastoris and purified with chromatography. By the facilitation of cholate, six kinds of rhapo penetrated among the phosphatidylcholine acyl chains. After purification by density-gradient centrifugation, rhHDL was acquired. Based on morphological observation, we confirmed that the micellar complexes of rhapo with phosphatidylcholine and cholesterol were prepared. We carried on comparative studies in vitro and in vivo between native HDL and rhHDL. Cellular cholesterol efflux assays showed that rhHDL could promote the efflux of excess cholesterol from macrophages. Furthermore, rhHDL has similar effects with native HDL on the blood lipid metabolism in hyperlipidemic mice. In conclusion, rhHDL has similar effects on antiatherosclerosis with native HDL through reverse cholesterol transport, antioxidative, and antithrombotic properties. It could be used as a therapeutic HDL-replacement agent.
