The Extent of Human Apolipoprotein A-I Lipidation Strongly Affects the β-Amyloid Efflux Across the Blood-Brain Barrier in vitro.

Roberta Dal Magro,Francesco Mantegazza,Gianvito Grasso,Laura Calabresi,Marco Agostino Deriu,Alysia Cox,Domenico Salerno,Sara Simonelli,Valeria Cassina,Francesca Re,Roberta Corti,Beatrice Formicola,Luca Nardo,Andrea Danani

doi:10.3389/fnins.2019.00419

Abstract

Much evidence suggests a protective role of high-density lipoprotein (HDL) and its major apolipoprotein apoA-I, in Alzheimer’s disease (AD). The biogenesis of nascent HDL derived from a first lipidation of apoA-I, which is synthesized by the liver and intestine but not in the brain, in a process mediated by ABCA1. The maturation of nascent HDL in mature spherical HDL is due to a subsequent lipidation step, LCAT-mediated cholesterol esterification, and the change of apoA-I conformation. Therefore, different subclasses of apoA-I-HDL simultaneously exist in the blood circulation. Here, we investigated if and how the lipidation state affects the ability of apoA-I-HDL to target and modulate the cerebral β-amyloid (Aβ) content from the periphery, that is thus far unclear. In particular, different subclasses of HDL, each with different apoA-I lipidation state, were purified from human plasma and their ability to cross the blood-brain barrier (BBB), to interact with Aβ aggregates, and to affect Aβ efflux across the BBB was assessed in vitro using a transwell system. The results showed that discoidal HDL displayed a superior capability to promote Aβ efflux in vitro (9 × 10-5 cm/min), when compared to apoA-I in other lipidation states. In particular, no effect on Aβ efflux was detected when apoA-I was in mature spherical HDL, suggesting that apoA-I conformation, and lipidation could play a role in Aβ clearance from the brain. Finally, when apoA-I folded its structure in discoidal HDL, rather than in spherical ones, it was able to cross the BBB in vitro and strongly destabilize the conformation of Aβ fibrils by decreasing the order of the fibril structure (-24%) and the β-sheet content (-14%). These data suggest that the extent of apoA-I lipidation, and consequently its conformation, may represent crucial features that could exert their protective role in AD pathogenesis.

Highlights

Apolipoprotein A-I is involved in the generation and metabolism of high-density lipoproteins (HDL) (Zannis et al, 2006)
Left smear represents the subclass of discoidal Apolipoprotein A-I (apoA-I)-HDL and right smear represents the subclass of spherical apoAI-HDL, as previously described (Didichenko et al, 2016)
This sample was treated with chymase in order to selectively degrade discoidal apoA-I-HDL, obtaining a preparation containing mature spherical apoA-I-HDL

Summary

Introduction

Apolipoprotein A-I (apoA-I) is involved in the generation and metabolism of high-density lipoproteins (HDL) (Zannis et al, 2006). The 28 kDa protein is synthesized in the small intestine and liver, and lipidated with cholesterol and phospholipids by the membrane-bound ATP-binding cassette transporter A1 (ABCA1) to create different subclasses of plasma HDL particles, including discoidal ones. During the HDL maturation process the secondary structure of apoA-I slightly changes (Sevugan Chetty et al, 2012). In humans, HDL consist of heterogeneous subclasses, characterized according to charge, size, density, protein, and/or lipid composition. HDL are involved in removing excess cholesterol from peripheral tissues, e.g., the arterial wall, and transporting it to the liver for secretion by reverse cholesterol transport. HDL exert other protective effects, including anti-oxidative, anti-inflammatory, anti-apoptotic, and anti-infective actions (Kingwell et al, 2014)

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