Polyunsaturated Fatty Acids Are Essential to Health Resolution of inflammation: Roles for resolvins and protectins

Bruce D. Levy Associate Professor of Medicine Pulmonary and Critical Care Medicine Brigham and Womens Hospital Harvard Medical School
ESPEN 2012 Barcelona New Horizons in Clinical Nutrition September 9, 2012 Disclosures: Licensed Patents Bayer Healthcare, Resolvyx Pharmaceuticals Consultant: Novartis, Eisai Pharmaceuticals
Thrombosis Hypertension Sudden death (GISSI study) Aspirin Allergies Asthma

Eye
Dry eye Macular degeneration

Omega-3 PUFA

Brain

EPA
Cardiovascular

DHA
Oral

Milligrams to grams daily

Brain functions Alzheimers Stroke Depression

Tooth loss Periodontal disease

Lungs

GI tract
Inflammatory Bowel Diseases (Colitis, Crohns disease)

Skin disorders Cancer

Immune systems Diabetes

Arthritis and Gout

Molecular mechanisms of action? Can EPA and DHA be precursors to generate bioactive mediators?

Wave Of Inflammation

Inflammation

Derek Willougby et al

Nature Immunology Reviews 2002

Resolution

Resolution Of Acute Inflammation

Ideal Outcome of Inflammation : Complete Resolution

Systems Approach to Mapping Resolution
Fibrosis
Chronic Inflammation
Chemical mediators: Prostaglandins Leukotrienes

Programmed Resolution

unresolved

Acute Inflammation

PGE2, D2, LTB4

Lipid mediator class switching

LX

Resolution

Return to homeostasis

Specialized Pro-Resolving Mediators C20:5

Resolvins
E-series
OH

C20:4

C22:6
Protectins

SPM Resolvins Protectins

Edema PMNs

Lipoxins
HO OH

COOH

D-series

OH

LXA4 RvE1

OH

HO COOH

OH OH

COOH

Monocyte/ Macrophages Activity

HO OH HO COOH

PD1

RvD1

Images from C. Fanta and L. Kobzik

sec-min

min hrs --days

time

Families Specialized Pro-Resolving Mediators (SPM): A Novel Genus Endogenous Lipid Mediators

Pro-Resolving Mediator Signaling Circuits

PUFA Mediator Receptor

Key Features in Tissue Resolution Catabasis

AA (C20:4) 15-LO 5-LO DHA (C22:6)

LXA4

ALX/FPR2

Anti-Inflammation
Block PMN recruitment Stop PMN Activation Decrease vascular permeability Dampen ROS

17S-RvD1 17R-RvD1 ASA-COX-2 5-LO

RvE1, RvD1, PD1

GPR32

Pro-Resolution
Stimulate endothelial NO and PGI2 Epithelial restitution Promote Mac phagocytosis of apoptotic PMNs, microbes and debris

RvE1, RvD1, PD1

EPA (C20:5)

RvE1

CMKLR1

NonNon-Lethal Experimental Model of ALI

Timed intervals
(2, 12, 48, 72h)

RvE1 Enhances Bacterial Clearance in a Model of Aspiration Pneumonia

Bacterial Growth Index 1.5 1.0

Harvest BAL
(Time)

Acid
(pH 1.5, 0.1 N, 50l) Anterior

0.5

Posterior

0 Treatment HCl

saline saline + + +

EPA RvE1 + + + +
J. Immunol. (2010);184:836-43.

E.coli
Mean SEM (n>12, each) *, P<0.05 vs saline / HCl (-) / E. coli (+) , P<0.05 vs saline / HCl (+) / E. coli (+)

Fukunaga et al. J. Immunol. 2005, 174:5033.

RvE1 Blocks Leukocyte Accumulation After Aspiration Pneumonia

1200 MPO (ng/ml) 800 400 0 RvE1 HCl + + + + +

RvE1 Improves Survival After Aspiration Pneumonia

Before ALI
HCl

RvE1 (-)

RvE1 (+)

After Pneumonia
E.coli
Observe survival for 1wk

HCl

E.coli

Observe survival for 1wk

40

30 min 12h RvE1 12h

2h RvE1

100

RvE1 saline

200

% Survival

% Survival

75 50 25 0

100 75 50 25 0 0

RvE1 saline

E.coli

Mean SEM (n>12, each) *, P<0.05 vs RvE1 (-) / HCl (-) / E. coli (-) , P<0.05 vs RvE1 (-) / HCl (+) / E. coli (+)

0 1 2 3 4 5 6 7 Time after E. coli inoculation (d)

1 2 3 4 5 6 7 Time after E. coli inoculation (d)

Mean SEM(n=10,each) *,P<0.05vs saline

J. Immunol. (2010);184:836-43.

J. Immunol. (2010);184:836-43.

Asthma Pathology

Resolvin Es Derived from EPA

Aspirin-independent pathway
Vascular Endothelial Cells
O(O)H 18R-hydroperoxy-EPE

P450 EPA
Leukocytes
COOH

Eicosapentanoic Acid EPA

Aspirin: COX2

HOOC

5-LOX

OH

OH

HO COOH

OH

5S(6)-epoxy18hydroxy-EPE

Resolvin E1

(5S,12R,18R-trihydroxy-EPE)

Healthy Individuals Taking EPA (1g) and Aspirin (160mg)

ng/ml plasma EPA with ASA (n=6) EPA w/o ASA (n=3) RvE1 0.18 0.06 0.11 0.02 18-HEPE 0.74 0.27 0.36 0.15 EPA 15.95 4.03 14.20 5.20
Serhan et al., J Exp Med. (2000) 192, 1197 Arita et al., J Exp Med. (2005) 201, 713

Provided by Robert Padera

Impact Of RvE1 On Airway Inflammation Resolution

RvE1

RvE1 Dampens Allergic Airway Responses Histopathology and Airway Reactivity

Vehicle H&E
Br Br Br 1.50 Br 1.25

0 Time (d) 2.5

14

15

16

17

18

19 20 21 Resolution

25

RvE1

AHR

OVA sensitization 1.50

OVA aerosol challenge

Vehicle 2.0 Total BALF cells ( 106) 1.25 RvE1 (100ng/d, iv)

400

MCh log ED200

V RvE1 V
4 Vehicle RvE1

1.00 0.75 0.50 0.25

1.5

BALF cells ( 106)

1.00

*
1.0

0.75

* *

BALF cells ( 106)

0.50

0.5

*
1 RvE1 Ri~2.5 days Vehicle Ri~5 days 0 0 18 21 Time (d) 25

400 Br Br

0.25

ic le Ve h

0.0
cl e R vE 1 Ve hi

0.00
Eo si no ph il s M ac ro ph ag es es yt oc ph

PAS
Representative of n > 6 Haworth et al. Nature Immunology. 2008; 9:873.

Protocol Day 21
n > 8, *P < 0.05

m Ly

Haworth et al. Nature Immunology. 2008; 9:873.

n > 6, *P < 0.05

vE 1

0.00

Resolvin Ds Derived from DHA

Docosahexanoic Acid DHA
LOX
H(O)O

AT-RvD1 Promotes Resolution of Allergic Airway Responses: Ag Clearance

ALX/FPR2 Expression: Epithelial Cells Leukocytes Macrophages

Leukocytes

COOH

17S-H(p)DHA

LOX 7S-hydroperoxy,17S-HDHA 7S(8)-epoxide intermediate 4S-hydroperoxy,17S-HDHA 4S(5)-epoxide intermediate

HO OH COOH OH

COOH

OH COOH OH HO

COOH OH OH OH

OH HO HO

Resolvin D1

Resolvin D2

Resolvin D3

Resolvin D4

Neutrophils Transendothelial migration Microglia IL-1 expression

Journal of Immunology. 2012. in press.

Serhan et al., 2002 J. Exp. Med. 196:1025-1037

Protectins Derived from DHA

Docosahexanoic Acid DHA
15-LOX-1
Human PBMC TH2-skewed PBMC

Protectin D1 Dampens Airway Inflammation

Lung histopathology
Vehicle Allergen-driven leukocyte infiltration

LOX

Brain, Microglial, retinal cells

H(O)O

COOH

O2
second oxygenation reduction

HO

17S-H(p)DHA
COOH

Cells in BALF (x105)

reduction enzymatic epoxydation

O

Vehicle PD1
(after aerosol challenge)

10S,17S-diHDHA
COOH

17S-HDHA

16(17)-epoxy-DHA enzymatic hydrolysis

Vehicle Neuroprotectin D1 Protectin D1 (PD1)

OH COOH OH

Eos

Lymphs

Serhan et al. J. Immunol. 2006;176:1848-1859.

J. Immunology 2007, 178:496-502.

Aerosol challenge of ovalbumin

Generation of Protectin D1 in asthma

Non-Invasive Technique For Sampling Airway Biomarkers
Exhaled Breath Condensates

d4-PGE2 i.s. J. Immunol. 2007, 178: 496-502.

Human EBC 17(S)-Hydroxy-DHA and PD1 Levels

Inflammation Resolution is a Natural Process

Healthy
17(S)-hydroxy-DHA PD1 Sample size (n) Trace 2.23 +/- 1.55 ng 3

Asthma Exacerbation
Trace Trace 4
In summary, we have presented evidence for: Resolution of inflammation is an active process that is orchestrated by specific cells and signals, including PUFA-derived lipid mediators. RvE1, RvD1 and PD1 are natural compounds, enzymatically derived from EPA and DHA that display pro-resolving actions Pro-resolving mediators (RvE1) enhance host defense

*Plus-minus

values are means +/- SEM.

d4-PGE2 i.s. Calibration curve r2 = 0.991 In conclusion, understanding the cellular and molecular mechanisms for pro-resolving mediators in catabasis is providing new insights into tissue responses for resolution of inflammation in health and the pathophysiology of sepsis and several other diseases of chronic inflammation with new opportunities for therapeutic intervention.

J. Immunol. 2007, 178: 496-502.

Acknowledgements

Pulmonary and Critical Care Medicine

Levy Lab Troy Carlo, Oliver Haworth, Michael Pfeffer, Anna Planaguma, Hiroyuki Seki, Mohib Uddin, Guangli Zhu, Jennifer Colby, Emiko Ono, Cindy Barnig, Alex de Paula Rogerio, Raja Abdulnour, Cindy Hana Okuma, Jack Timmons Asthma Research Center - Elliot Israel, Gautham Marigowda, Shamsah Kazani Michael Wechsler, Christopher Fanta

Center for Experimental Therapeutics and Reperfusion Injury

Charles N. Serhan, Director Sungwhan Oh, Fei Gao

Funding: NHLBI, NIAID, NIGMS