A SEMINAR PRESENTATION ON

REVERSE CHOLESTEROL TRANSPORT

OUTLINE
ACKNOWLEDGEMENT INTRODUCTION CHOLESTEROL OVERVIEW CHOLESTEROL TRANSPORT HDL BIOCHEMISTRY REVERSE CHOLESTEROL TRANSPORT (RCT) TRANSPORT PROTEINS IN RCT CLINICAL SIGNIFICANCE OF RCT CONCLUSION REFERENCES

ACKNOWLEDGEMENT
Firstly, I acknowledge God Almighty for his grace and enablement to commence and complete this report. I appreciate the effort and selfless nature of my supervisor. I appreciate the Head of Department, Faculty and Staff of the Department of Biological Sciences, Covenant University, who have also contributed significantly to my knowledge in Biochemistry. I also appreciate my Parents, for their love and support.

INTRODUCTION
About 40 years ago, John Glomset outlined the reverse cholesterol hypothesis. Glomset proposed that HDL and LCAT might have antiatherogenic functions related to their ability to transport cholesterol from peripheral tissue to the liver for excretion (Tall, 2003). In order to dispose of cholesterol, it is transported to the liver and intestine from periphery and is finally excreted via the feces.

 This pathway has been traditionally referred as reverse cholesterol transport or centripetal cholesterol flux. Both free cholesterol and the esterified form are involved in RCT. Excess unesterified cholesterol is toxic to cells, and therefore, cells have developed several ways to protect themselves against cholesterol toxicity. The return of this peripheral cholesterol to the liver is necessary to balance cholesterol intake and de novo synthesis and thus to maintain whole body steady-state cholesterol metabolism (Cuchel and Rader, 2006).

What is cholesterol?
Cholesterol is a waxy, fat-like substance that is found in all cells of the body.

Cholesterol is an amphiphatic lipid.

Cholesterol is derived from dietary source and de novo synthesis.

PROPERTIES
Its molecular formula is C27H46O. Its an organic compound. It has a melting point of 148-150 0C It has a boiling point of 360 0C

STRUCTURE
cyclopentanoperhydrophenanthrene ring

IUPAC name: (3)-cholest-5-en-3- ol.

Figure.1. Cholesterol Structure (Sterols: Cholesterol and cholesterol esters. http://lipidlibrary.aocs.org/lipids/cholest/index. htm. Sourced on Sunday, 22 September 2013 at 5p.m.).

FUNCTIONS
It plays a dual role as an essential structural component of cellular membranes and a regulator of gene transcription, protein degradation, and enzyme activity (Liscum and Dahl, 1992). It is required to build and maintain membrane. It regulates membrane fluidity over the range of physiological temperature. Within the cell membrane, cholesterol also functions in intracellular transport, cell signaling and nerve conduction. It is a precursor for the synthesis of vitamin D and steroid hormones.

SYNTHESIS
The major site of synthesis of cholesterol are Liver Adrenal cortex Testes Ovaries and Intestine occurs in the endoplasmic reticulum and the cytosol

STEPS:
I. Synthesis of mevalonate occurs from acetylCoA. II. Formation of Isoprenoid units from mevalonate and loss of CO2. III. Condensation of isoprenoid units to form squalene. IV. Cyclisation of Squalene to the parent steroid (lanosterol). V. Cholesterol formation from lanosterol.

Figure.2. Cholesterol biosynthetic pathway (Vance and Vance, 2002).

CHOLESTEROL TRANSPORT
Blood is watery and cholesterol is fatty. Just like oil and water, the two do not mix. So, in order to travel in the bloodstream, cholesterol is carried in small packages called lipoproteins. The small packages are made of fat (lipid) on the inside and proteins on the outside.
WHAT THEN ARE LIPOPROTEINS?

 Lipoproteins are complex aggregates of lipids and proteins that renders the lipids compatible with the aqueous environment of the body fluids and enable their transport throughout the body of all vertebrates to tissues where they are required. Two kinds of lipoproteins carry cholesterol throughout the body.

Low density lipoprotein (LDL) High density lipoprotein (HDL)

LDL
LDL carries cholesterol from the liver to other tissues (extra-hepatic tissues). LDL is absorbed by target cells through receptor mediated endocytosis (RME). A high LDL cholesterol leads to build up of cholesterol in the arteries.

HDL
HDL carries cholesterol from other part of the body back to the liver. The liver then removes the cholesterol from the body. HDL in the liver is not taken up by RME, rather they dock in the surface receptor, deposit its cholesterol. And depart as remnant without being incorporated into the cells interior.

BIOCHEMISTRY OF HDL
HDL carries about 33% of plasma cholesterol. Since it is associated with efflux and redistribution of cholesterol in extra-hepatic tissues, it is thought to play a prominent role in RCT. HDL has a density of 1.063g/ml. It is made of mostly proteins and small amount of cholesterol. The protein component (apoprotein) includes;

 Ap0 A-1 Ap0 A-2 Apo C Apo D Apo E

Apo A-1 is quantitatively and functionally the most important (Berger, 1984). It makes up 70% of HDL protein and found in every HDL particle. Freshly formed or secreted HDL exists briefly in the form of bilayered discs containing a high proportion of protein (mainly apo A-I) and phospholipid relative to cholesterol.

 Lipid poor Apo A-1 (bilayer disc form of HDL) interacts with ABCA1 on macrophage form cells, forming nascent HDL particle. The nascent particle interacts with ABCG1 and SR-B1 transporter to develop into a mature HDL particle (Berger, 1984). At the present time the most convenient clinical subdivision of HDL is into HDL2 and HDL3. HDL2 is larger and less dense than HDL3 enriched in cholesterol ester. Matured HDL delivers cholesterol to the liver through : Direct and Indirect pathways.

REVERSE CHOLESTEROL TRANSPORT

RCT is a pathway for plaque reduction (Cuchel and Rader, 2006). From macrophages, cholesterol can be efflux as free cholesterol either via ATP binding cassette transporter A1 (ABCA1) with poor lipidated Apo A-1 as acceptor or via ABCG1 with more mature spherical HDL particles serving as receptor (Linsel and Tall, 2005). Additional efflux capacity might be provided by scavenger receptor class B type 1 (SR-B1) or by aqueous diffusion (Wang and Rader, 2007).

 Within HDL, cholesterol is esterified by lecithincholesterol acyl-transferase (LCAT). This is done so to facilitate more uptake of free cholesterol. Via the plasma compartment the effluxed cholesterol is transported in a reverse pathway back to the liver. HDL-derived cholesterol is then de-esterified and secreted into the bile

INDIRECT PATHWAY DIRECT PATHWAY HDL interacts with SR-B1 (scavenger receptor B1) on the liver allowing cholesterol delivery. The lipid-poor HDL particle can be re-circulated to repeat the process of RCT. This is mediated by CETP (cholesteryl ester transfer protein). CETP facilitates the exchange of cholesterol in HDL for triglyceride in triglyceride-rich apo B particle: VLDL and LDL. HDL is enriched with triglyceride and LDL with cholesterol. LDL particle may go into circulation or interact with LDL-receptors at the liver cells and deposit the cholesteryl ester.

Cholesterol lacks enzyme system which can break the steroid nucleus of cholesterol. So it is not degraded. It is however converted to bile acid in the liver and eliminated through the bile.

Figure.3. A schematic diagram depicting the role of lipases in reverse cholesterol transport (Rader, 2003).

TRANSPORT PROTEINS INVOLVED IN RCT

ABCA1: Facilitates the efflux of unesterified cholesterol and phospholipid. Genetic deficiency of ABCA1 causes Tangier disease. LCAT: Necessary for the formation of matured HDL and for remodeling of HDL lipoprotein particle (Dobiasova and Frohlich, 1999). Traditionally, LCAT activity has been considered anti-atherogenic. Its protective role depends on i. concentration and quality of plasma HDL ii. LDL particles iii. availability of lipid transferring protein

 SR-B1: Mediates selective uptake of HDL cholesterol by the liver . SR-B1 is expressed in macrophages and may contribute to cholesterol efflux from macrophages under certain conditions (Dobiasova and Frohlich, 1999).

CETP: mediates transfer of cholesteryl ester from HDL to apolipoprotein B-containing lipoproteins in exchange for triglyceride (Tall, 2003).

CLINICAL SIGNIFICANCE OF RCT

Early diagnosis of hypercholesterolemia (which promotes atheroma and leads to myocardial infarction, stroke and peripheral vascular disease). Components of RCT useful in clinical trial includes; i. Enzyme activity ii. Transport proteins iii. Membrane modulators iv. Apo-proteins v. HDL classes.

CONCLUSION
From the different steps that are important in the RCT pathway, overall RCT might be differentially affected on different levels which includes; i. The macrophage ii. Transport of cholesterol through the plasma compartment iii. The uptake by the liver iv. The excretion into the intestine, and v. The excretion from the body (Annema and Tietge, 2012).

REFERENCES
Annema, W. and Tietge, J.F. (2012). Regulation of reverse cholesterol transport: A comprehensive appraisal of available animal studies. Nutrition & Metabolism 9: 1-18. Berger, G.M.B. (1984). High-density lipoproteins, reverse cholesterol transport and atherosclerosis-recent developments. South African Medical Journal 65: 503-506.

Colpo, A. (2005). LDL cholesterol: BAD cholesterol or BAD science?. American Journal of physicians and surgeons 10: 83-87.
Cuchel, M. and Rader, D.J. (2006). Macrophage reverse cholesterol transport: Key to the regression of atherosclerosis? Circulation 113: 2548-2555. Dobiasova, M. and Frohlich, J.J. (1999). Advances in understanding of the role of lecithin cholesterol acyltransferase (LCAT) in cholesterol transport. Clinica Chimica Acta 286: 257-258.

Mayes, P.A. and Botham, K.M. (2003). Cholesterol Synthesis, Transport and Excretion. In: Harpers illustrated biochemistry. (26). Lange Medical Books/McGraw-Hill., pp: 219-220. ISBN-0-07-138901-6. Linsel-Nitschke, P. and Tall, A.R. (2005). HDL as a target in the treatment of atherosclerosis cardiovascular disease. Nature Reviews Drug Discovery 4: 193-205.
Liscum, L. and Dahl, N.K. (1992). Intracellular cholesterol transport. Journal of lipid research 33: 1239-1240. Rader, D.J. (2003). Regulation of reverse cholesterol transport and clinical implications. American Journal of Cardiology 92: 42J-49J. Tall, A.R. (2003). Role of ABCA1 in cellular cholesterol efflux and reverse cholesterol transport. Arteriosclerosis Thrombosis Vascular Biology 23: 710-711. Wang, X. and Rader, D.J. (2007). Molecular regulation of macrophage reverse cholesterol transport. Current Opinion in Cardiology 22: 368-372.

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