1Bruce D. Levy
Associate Professor of Medicine Pulmonary and Critical Care Medicine
Brigham and Womens Hospital
Harvard Medical School
ESPEN 2012 BarcelonaNew Horizons in Clinical Nutrition
September 9, 2012
Disclosures: Licensed Patents Bayer Healthcare, Resolvyx PharmaceuticalsConsultant: Novartis, Eisai Pharmaceuticals
Resolution of inflammation: Roles for resolvins and protectins
Cardiovascular
Aspirin
Thrombosis Hypertension Sudden death(GISSI study)
Molecular mechanisms of action?Can EPA and DHA be precursors to generate bioactive mediators?
Polyunsaturated Fatty Acids Are Essential to Health
EPA DHA
Omega-3 PUFA
Milligrams to grams daily
BrainEye Brain functions Alzheimers Stroke Depression
Dry eye Macular degeneration
DiabetesCancerArthritis and Gout
GI tractLungs
Skin disorders
Inflammatory Bowel Diseases(Colitis, Crohns disease)
Allergies Asthma
Immune systems
Oral
Tooth loss Periodontal disease
Derek Willougby et al Nature Immunology Reviews 2002
Wave Of Inflammation
InflammationResolution
2Images from C. Fanta and L. Kobzik
Resolution Of Acute Inflammation
Lipoxins ProtectinsResolvins
D-seriesE-series
COOH
OH
OH
OH
COOH
HOOH
RvE1
LXA4
PD1
HO OH
OH
COOH
COOH
HO
HO
OH
RvD1
Specialized Pro-Resolving Mediators
Monocyte/Macrophages
PMNs
Act
ivity
Edema
SPMResolvinsProtectins
timeFamilies Specialized Pro-Resolving Mediators (SPM):
A Novel Genus Endogenous Lipid Mediators
Programmed Resolution
Ideal Outcome of Inflammation : Complete Resolution Systems Approach to Mapping Resolution
Resolution
FibrosisChronic
Inflammation
Acute Inflammation
unresolved
Lipid mediator class switching
Chemical mediators:ProstaglandinsLeukotrienes
PGE2, D2, LTB4 Return to homeostasisLX
sec-min min hrs --days
C20:4 C20:5 C22:6
Pro-Resolving Mediator Signaling Circuits
AA (C20:4)
DHA (C22:6)
EPA (C20:5)
LXA4
17S-RvD117R-RvD1
RvE1
ALX/FPR2
GPR32
CMKLR1
ASA-COX-25-LO
15-LO5-LO
PUFA Mediator Receptor
Key Features in Tissue ResolutionCatabasis
Anti-InflammationBlock PMN recruitmentStop PMN ActivationDecrease vascular permeabilityDampen ROS
RvE1, RvD1, PD1
Pro-ResolutionStimulate endothelial NO and PGI2Epithelial restitutionPromote Mac phagocytosis of apoptotic PMNs, microbes and debris
RvE1, RvD1, PD1
3Acid(pH 1.5, 0.1 N, 50l)
(Time)
HarvestBALTimed intervals
(2, 12, 48, 72h)
PosteriorAnterior
NonNon--Lethal Experimental Model of ALILethal Experimental Model of ALI
Fukunaga et al. J. Immunol. 2005, 174:5033.
Treatment saline EPA RvE1salineHCl - + + +
E.coli + ++ +
*
0
0.5
1.0
1.5
Bacte
rial G
rowth
Index
RvE1 Enhances Bacterial Clearance in a Model of Aspiration Pneumonia
Mean SEM (n>12, each)*, P
4Asthma Pathology
Provided by Robert Padera
5S(6)-epoxy-18hydroxy-EPE
OOH
COOH
5-LOX Leukocytes
HOOC
O(O)HVascular Endothelial Cells
Aspirin: COX2
18R-hydroperoxy-EPEEicosapentanoic AcidEPA
Resolvin E1
OH
COOH
HOOH
Resolvin Es Derived from EPA
(5S,12R,18R-trihydroxy-EPE)
Serhan et al., J Exp Med. (2000) 192, 1197Arita et al., J Exp Med. (2005) 201, 713
EPAP450
Aspirin-independent pathway
Healthy Individuals Taking EPA (1g) and Aspirin (160mg)
ng/ml plasma RvE1 18-HEPE EPA
EPA with ASA (n=6)
0.18 0.06 0.74 0.27 15.95 4.03
EPA w/o ASA (n=3)
0.11 0.02 0.36 0.15 14.20 5.20
RvE1
0 7 2514 1815 16 17 19 20 21
OVA aerosol challenge
Time (d) OVA sensitization Resolution
Vehicle Ri~5 days
*
BALF
cel
ls (
106 )
0.00
0.25
0.50
0.75
1.00
1.25
1.50
*
RvE1 (100ng/d, iv)
Vehicle
*
Lymp
hocy
tes
Macro
phag
es
Eosin
ophil
s
Impact Of RvE1 On Airway Inflammation Resolution
RvE1 Ri~2.5 days
0
1
2
3
4
18 21 25
*
Time (d)
BALF
cel
ls (
106 )
VehicleRvE1
0
RvE1
Tota
l BAL
F ce
lls (
106 )
0.0
0.5
1.0
1.5
2.0
2.5
*
Vehic
le
RvE1
Protocol Day 21n > 8, *P < 0.05 Haworth et al. Nature Immunology. 2008; 9:873.
400
400
Br Br
VV
Br Br
BrBr
Vehicle RvE1
RvE1 Dampens Allergic Airway ResponsesHistopathology and Airway Reactivity
H&E
PAS
Representative of n > 6
0.00
0.25
0.50
0.75
1.00
1.25
1.50
MC
h lo
g ED
200
*
Vehic
leRv
E1
n > 6, *P < 0.05
AHR
Haworth et al. Nature Immunology. 2008; 9:873.
5Resolvin Ds Derived from DHA Docosahexanoic Acid
DHA
17S-H(p)DHA
LOX
Resolvin D1 Resolvin D2 Resolvin D3 Resolvin D4
Leukocytes
COOH
H(O)O
4S(5)-epoxide intermediate
COOH
HO
HO
OH
HO
OH
COOHOH
COOH
OH
OH
HO
OH
OH
COOH
OH
4S-hydroperoxy,17S-HDHA
7S(8)-epoxide intermediate
7S-hydroperoxy,17S-HDHA
Serhan et al., 2002 J. Exp. Med. 196:1025-1037
LOX
Neutrophils Transendothelial migration
Microglia IL-1 expression
AT-RvD1 Promotes Resolution of Allergic Airway Responses: Ag Clearance
ALX/FPR2 Expression:Epithelial Cells
LeukocytesMacrophages
Journal of Immunology. 2012. in press.
Docosahexanoic AcidDHA
17S-H(p)DHA
LOX Brain, Microglial, retinal cells
COOH
H(O)O
COOH
O
16(17)-epoxy-DHA
17S-HDHA
COOH
H Oreduction
enzymatic epoxydation
enzymatic hydrolysis
second oxygenation
O2
10S,17S-diHDHA
Neuroprotectin D1Protectin D1 (PD1)
Human PBMC TH2-skewed PBMC
15-LOX-1
reduction
Protectins Derived from DHA
COOH
OH
OH
Serhan et al. J. Immunol. 2006;176:1848-1859.
Protectin D1 Dampens Airway Inflammation
Lung histopathology
Vehicle
Aerosol challenge of ovalbumin
Allergen-driven leukocyte infiltration
Vehicle Eos Lymphs
Cel
ls in
BAL
F (x
105 )
VehiclePD1 (after aerosol challenge)
J. Immunology 2007, 178:496-502.
6Non-Invasive Technique For Sampling Airway BiomarkersExhaled Breath Condensates
Generation of Protectin D1 in asthma
d4-PGE2 i.s.
J. Immunol. 2007, 178: 496-502.
d4-PGE2 i.s.
Calibration curve r2 = 0.991
Human EBC 17(S)-Hydroxy-DHA and PD1 Levels
43Sample size (n)
Trace2.23 +/- 1.55 ngPD1
TraceTrace17(S)-hydroxy-DHA
Asthma ExacerbationHealthy
*Plus-minus values are means +/- SEM.
J. Immunol. 2007, 178: 496-502.
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
In conclusion, understanding the cellular and molecular mechanisms for pro-resolvingmediators in catabasis is providing new insights into tissue responses for resolution of inflammation in health and the pathophysiology of sepsis and several other diseases ofchronic inflammation with new opportunities for therapeutic intervention.
Inflammation Resolution is a Natural Process
7Acknowledgements
Center for Experimental Therapeutics and Reperfusion InjuryCharles N. Serhan, DirectorSungwhan Oh, Fei Gao
Pulmonary and Critical Care MedicineLevy 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 KazaniMichael Wechsler, Christopher Fanta
Funding:NHLBI, NIAID, NIGMS