Cardiovascular Disease and Inflammation: What Can HDL Do ... … · Cell Membrane Translocation...

Alan T. Remaley, MD, PhD

Lipoprotein Metabolism SectionCardiovascular and Pulmonary Branch

National Heart, Lung and Blood InstituteNational Institutes of Health

Conflicts of Interests: Research grants with MedImmune, Shire, Corvidia, and Liposcience

Cardiovascular Disease and Inflammation: What Can HDL

Do About It?

Overview

• History of Cholesterol Research

• Lipoprotein Metabolism

• Pathogenesis of Atherosclerosis

• HDL and Inflammation

History of Cardiovascular Disease Research

Rudolf Virchow 1821-1902

H. Anitchkov1885-1964

Cholesterol

(factor in CHD)

LDL

(Bad - chol)

HDL

(neutral)

LDL

(Bad - chol)

HDL

(Good - chol)

Ox-LDL

(Very Bad - chol)

LDL

subfractions

Non-HDL

Lipoproteins

Dysfunctional

HDL

Bad HDL

Good HDL

1905 1920’s-1950’s 1970’s 1980’s 1990’s 2000’s

Evolution of Cholesterol as a CHDRisk Biomarker

Early Lipoprotein Research at the NIH

Circa mid 1960s

Overview





Chylomicron

VLDL

IDL

LDL

HDL

Lp(a)

CholesterolProteinTriglyceridePhospholipid

Major Classes of Lipoproteins

Lipoprotein Metabolism Pathways


Triglyceridedelivery

for energy metabolism


Cholesterol Removal

HDL Structure and Composition

Pre-beta Discoidal Shaped HDL

Alpha Spherical Shaped HDL

Initial Cholesterol Acceptor Delivers Cholesterol to Liver

HDL transports over 80 different

proteins and over 200 species

of lipids.

Pleiotropic Anti-AtherogenicEffects of HDL

Antiinflammatory

Activity

Reverse

Cholesterol

Transport

Antithrombotic

Activity

Antiapoptotic

Activity

Antioxidative

Activity

Antiinfectious

Activity

Epithelial

Repair

Vasodilatory

Activity

Curr Med Res Opin 2004:20:1253-1268

Annu Rev Med 2003:53:321-341

Antiinflammatory

Activity

Reverse

Cholesterol

Transport

Antithrombotic

Activity

Antiapoptotic

Activity

Antioxidative

Activity

Antiinfectious

Activity

Epithelial

Repair

Vasodilatory

Activity

Curr Med Res Opin 2004:20:1253-1268

Annu Rev Med 2003:53:321-341

Pleiotropic Anti-AtherogenicEffects of HDL

Reverse Cholesterol Transport Pathway


Cholesterol Esterification

C

LIVER

LCAT

DiscoidalHDL

Diffusion

CellMembrane

TranslocationStep

SphericalHDL

Dissociation

KIDNEY

Free Apo A-I

HL

CETP

ABCAI“Flippase”

2 ATP 2 ADP

Apo A-IBinding Site

Apo A-IBinding Site

C

Association

Apo A-I

C

Secretion

C

C

CE

Model of Cholesterol Efflux to HDL

Overview





M-CSFMCP-1

VCAM-1

Early Inflammatory Mediatorsof Atherosclerosis

LDL Uptake

Lumen

LipidCore

Fibrous cap

Shoulder

Intima

MediaElasticlaminæ

Internal

External

Calcification

Features of a Complex Atherosclerotic Plaque

No symptoms + Symptoms

Time (y)

Symptoms

Lesion initiation

Ischemic HeartDisease

CerebrovascularDisease

Peripheral VascularDisease

Schematic Timeline for Atherogenesis

Ruptured Atherosclerotic Plaque

Thrombus

Necrotic LipidCore Ruptured

FibrousCap

ThickenedMedia

100 160 220

HDL-C is an Independent CVD risk factor

HDL-C

(mg/dL)LDL-C (mg/dL)

25

Gordon T et al. Am J Med 1977;62:707-714.

4565

85

”Bad Cholesterol”

”Good Cholesterol”

Multiple Anti-atherogenic Effects of HDL

HDL

LDL

LDL

Ox

LDL

Inhibition of

LDL-oxidation

Inhibition of

monocyte-

adhesion

Cholesterol-

efflux

HDL

HDL

HDL

Endothelium

mono-

cyte

macro-

phagefoam

cell

Inhibition of

LDL Infiltration

Modulation of LDL-C versus HDL-C for CHD Risk Reduction

First Major Crack in HDL Hypothesis

Torcetrapib

Illuminate Phase 3 trialhalted for increased CVD events

Forbes, April 2007 Bartrer P et al. NEJM 2007

HDL-C: 72% increaseLDL-C: 25% decrease

Overview





Potential Anti-inflammatory Mechanisms of HDL

• Cholesterol Removal-Both cellular and extracellular

• Sequestration of oxidized lipids/LPS/cytokines

• Delivery of Anti-inflammatory Proteins and Lipid Cargo

5A peptide

Alpha-helix

cross-section

Kidney Int. 89 (2106) 809JIMD Rep. 2 Dec (2016)J Lipid Res. 56 (2015) 1727PLoS One. 8 (2013) e68802J Biol Chem. 287 (2012)43730J Pharm Exp Ther. 344 (2013) 50Am J Respir Cell Mol Biol. 47 (2012) 186

5A ApoA-I Mimetic Peptide

5A-POPCControl

5A Inhibits Cytokine Release by

Macrophages

Anna Schwendeman et al. J. Lipid Res. 2015;56:1727-1737

TNF-alpha release murine

macrophagesCytokine release PHA-treated

whole blood

Ctrl-hatched

5A-POPC-open

5ASM-grey

LpX

Bilayer or multi-lamellar

complex of phospholipids

Aqueous core

Features of LCAT Deficiency

Cloudy Cornea Renal Lipid Deposits

Partial deficiency: Fish Eye Disease (FED)• Cloudy cornea

Complete deficiency: Familial LCAT Deficiency (FLD)• Cloudy cornea• Normochromic normocytic anemia• Proteinuria/ESRD• Low HDL-C• Presence of LpX

Synthetic Fluorescent LpX

Agarose gelPlasma PE fluorescence

LpX Causes Renal Disease in LCAT Deficiency

Plasma Clearance of LpX

is Delayed in Lcat-/- Mice

-origin

PLoSOne 26 (2016) e0150083

Synthetic Fluorescent LpX

Agarose gelPlasma PE fluorescence

LpX Causes Renal Disease in LCAT Deficiency

WT

Lcat-/-

Glomerular Deposition of LpX

Basement membrane

Mesangium

Plasma Clearance of LpX

is Delayed in Lcat-/- Mice

-origin

PLoSOne 26 (2016) e0150083

LpX Induces Podocyte Effacement

TEM SEM

LpX Causes Glomerulosclerosis

LpX Causes Podocyte Effacement and Proteinuria

LpX Causes Proteinuria

LCAT-KO

WT

PLoSOne 26 (2016) e0150083

0

10

20

30

40

50

60

70

80

90

Baseline week 2 week 3 week 4 week 5

Uri

ne

Alb

um

in t

o C

reat

inin

eR

atio

(u

g/m

g)

Time

LCAT-KO + LPx

WT + LpX

LCAT-KO + saline

LCAT-KOSaline

LpX Activates the Inflammasome and Fixes Complement

42

L p -X

N o c h o le s te r o l

L p -X

C h o le s te r o l

0

2 0 0 0

4 0 0 0

6 0 0 0

8 0 0 0

C3

a n

g/m

l

*

Un

treate

d

Lp

X25

Lp

X33

Lp

X50

Lp

X65

0

1 0 0

2 0 0

3 0 0

M o n o c y te D e r iv e d M a c r o p h a g e s

IL-1

p

g/m

l

**

**

Cholesterol content

of Lp-X

% D

es

tab

liz

ed

Ly

so

so

me

Un

treate

d

Lp

X

Lp

X H

DL

0

5

1 0

1 5

**

C 3

b

0 2 4.5 8.5 30mol% cholesterol

IL-1 Secretion C3 Fixation

Lysosome Destabilization C3 Fixation

Complement Fixation of CholesterolCholesterol

Cholestane

C3 Fixation

Cholesterol crystals Induce Inflammation

• Crystals can be seen as early as 2 weeks

NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals. Peter Duewell Eicke Latz

H and E Crystals Hoechst Macrophages

Immunol 149 (2016) 306

HDL Suppresses InflammasomeActivation by Cholesterol Crystals

45 of 22

Immunol 149 (2016) 306

HDL Dampens LPS Inflammatory Response

ATVB 27(2007) 1153

https://microbeonline.com/lipopolysaccharide

HDL-Proteome

J Clin Invest 2007; 117: 746

Functional Map Interactome Map

Rosuvastatin Increases A1AT on HDL

Mol Cell Proteomics 14 (2015) 3247

Molecular Modeling

A1AT (Red), HDL (Green)

LipidInterface

Cryoelectron Microscopy

Protein Spectral Count Changes

LipidInterface

Hphob.Loop

Role of PAR in Inflammation

Mol Cell Proteomics 14 (2015) 3247

Elastase Induced Inflammation HDL Delivers A1AT to Lung

A1AT-HDL Inhibits Elastase-induced Inflammation

Am J Resp Cell Mol Biol 51 (2015) 4

Protease Activated ReceptorsProposed Role of A1AT-HDL in Atherogenesis

HDL-Lipidome: Major Lipid Classes

SterolsHDL

>200 species of specific lipids

Akt

HDLS1P1/S1P3

Ga

eNOS

NO

Vasodilation

SR-B1

N C

A-I

g

S1P

Migration

Rac1

BAD

Bcl-XL

PP

Caspase 9/3

Apoptosis

NF-kB

E-selectin

VCAM-1

Adhesion

p38MAPK

COX-2

PGI2

TGF-2

Ga

g

Smad2/3

MMP9PLA2

Inflammation

RhoA

S1P2

Rac1

Smad2/3NADPH

oxidase

ROS

HDL-S1P Inversely Related CHD

Subjects with CHD have decreased S1P on HDL.

Lipids Health Disease 2011; 10: 70

S1P Content of HDL

HDL-S1P Inversely Related CHD

Subjects with CHD have decreased S1P on HDL.

Lipids Health Disease 2011; 10: 70

S1P Content of HDL Endothelial Integrity

S1P on HDL improves endothelial barrier function.

Major Questions in HDL Field• Is it involved in the pathogenesis of CVD?

• Should we measure it?

• Is it a target of therapy or just a CVD biomarker?

• What is the future of HDL research?Composition studies and how relate to function.

Most likely, but maybe not RCT.

Yes, but need better assays.

No for HDL-C, but can be under some conditions.

HDL research is still valuable despite recent clinical trial failures and more research not less is needed to harness its potential.

National Institutes of HealthMarcelo Amar

Lita Freeman

Ed Neufeld

Nori Hosaka

Takafumi Nishida

Rafique Islam

Milton Pryor

Aki Sakurai

Toshi Sakurai

Lufan Sun

Denis Sviridov

Robert Shamburek

Mike Stagliano

Jingrong Tang

Boris Vaisman

Ania Wolska

Zhihong Yang

Contact Information

Alan T. Remaley, MD, PhD

National Institutes of Health

NHLBI

Bethesda, MD

[email protected]

Cardiovascular Disease and Inflammation: What Can HDL Do ... … · Cell Membrane Translocation...

Documents

Transcript of Cardiovascular Disease and Inflammation: What Can HDL Do ... … · Cell Membrane Translocation...