Download A. The LIPID MAPS Initiative B. Example: Macrophage Eicosanoids

PERCEPTION OF THE LIPIDOME AFTER THE
FIRST DECADE OF LIPIDOMICS
A. The LIPID MAPS Initiative
B. Example: Macrophage Eicosanoids
D. Human Plasma Lipidome and Biomarkers
Edward A. Dennis
Departments of Chemistry/Biochemistry and Pharmacology
School of Medicine, University of California, San Diego
LIPID MAPS “Glue Grant” NIH U54 GM 69338
4 base side chains
20 amino acid side chains
Lipidomics Joins
the Omics Evolution
(2009), Dennis,
PNAS, 106, 2089.
nucleic acids, amino acids,
sugars and fats: >105 ?
all the fats: >105 ?
Number of Citations
Increase in Omics Citations 1985-2009
Wenk MR (2010) Lipidomics:
New Tools and Applications.
Cell, 143, 888-895.
Year
Lipidomics Publications 2002-2013
Lipidomics publications
Publications
Publications
LIPID MAPS publications
Year
Year
_____________
LIPID MAPS
TM
TM
Saccharolipids / Polyketides
_______________
“CLASS”: Comprehensive
Lipidomics Analysis using
Separation Simplification
cells or tissues
probe
cells
tissues
medium
sonicate
a “divide-and-conquer” strategy
homogenate
category specific
internal standards
(deuterated, odd-chain carbon)
category optimized
extraction
(liq-liq, SPE)
extract
GC
category specific
LC
(GC, NP-HPLC, RP-HPLC, chiral, specialty)
mass spectrometer
Harkewicz & Dennis, “Applications of mass
spectrometry to lipids and membranes”
Ann Rev Biochem, 80: 301-325 (2011)
(variables)
1. Mass spectrometer types
2. Ionization mode
3. Additives (for ionization)
4. Mass spectrometer monitoring modes
Kdo2-Lipid A (KLA, LPS subspecies)
A specific agonist of TLR-4 on RAW 264.7 macrophages
Raetz et al., 2006, J. Lipid Res. 47: 1097
Nuclei – DAPI
Mitochondria – MitoTracker Red
O-Specific
chain
Polysaccharide
Core
Lipid A
Glycophospholipid
Kdo
Dennis et al (2010)
J. Biol. Chem, 51, 39976-85
Cellular Eicosanoid Metabolism
Buczynski, Dumlao, Dennis (2009) JLR, 50, 1015-38
Cellular eicosanoid metabolism
CYP(ω)
COX
CYP(EET)
15-LOX
12-LOX
Buczynski, Dumlao, Dennis, 2009, JLR, 50: 1015-38
5-LOX
Cellular eicosanoid metabolism
COX
Directed Proteomics on Enzymes of
Eicosanoid Biosynthesis
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Sabido, Quehenberger
Dennis, Aebersold
(2012)
Mol Cell Proteomics 11:
M111.014746
Eicosanoid Genes to Proteins to Metabolites
Proteins
Quehenberger & Dennis, New Eng. J. Medicine, 365, 1812-23 (2011)
The Beneficial Lipids in Fish Oil
•  Fish oil is composed primarily of two ω-­‐3 PUFAs: ω-­‐3 ω Eicosapentaenoic Acid (EPA) (C20:5, ω-­‐3) ω-­‐3 ω Docosahexaenoic Acid (DHA) (C22:6, ω-­‐3) •  Humans synthesize from essen=al fa>y acid α-­‐linolenic acid (C18:3, ω-­‐3) •  EPA is found at barely detectable levels in humans •  DHA is more abundant than EPA; found primarily in the brain and re=na Arachidonic Acid Serves as a
Precursor for Many Eicosanoids
?
?
?
EPA/DHA
?
?
Are eicosanoids less pro-inflammatory when derived from EPA/DHA?
Effects of PUFA Supplementation on ATP
Stimulated COX-1 and 5-LOX Signaling
Norris & Dennis, PNAS (2012) 109, 8517
Omega-3 Implications for Nutrition
•  The global effects of EPA and DHA on normal lipid
metabolism can be quantitatively studied.
•  EPA and DHA affect the overall eicosadome decreasing
production of some, but not all, AA-derived eicosanoids.
•  There is a concomitant increase in specific EPA- and
DHA-derived metabolites.
•  Deciphering the role of fish oil-derived ω-3 EPA and DHA
in inflammatory eicosanoid signaling provides insight as
to their role as therapeutic/nutritional agents.
Norris & Dennis, PNAS (2012) 109, 8517
NIST collected (pooled) fasting plasma from 100 individuals
50% female and 50% male; age 40-50
15% of the total taken from
individuals of Hispanic origin!
Human Plasma Metabolites (mg/dL)
Lipids
Nucleic Acids
Amino Acids
Sugars
Human Plasma Lipid Categories (µM)
Sterol Lipids
Sphingolipids
Fatty Acyls
Glycerolipids
Glycerophospholipids
Prenols
Human Plasma Lipid Diversity
J Lipid Res, 51, 3299-3305 (2010)
Plasma Lipids in the Metabolic Syndrome
Quehenberger & Dennis, New Eng. J. Medicine, 365, 1812-23 (2011)
Charles Brown
Univ. of Missouri
George Kokotos
Univ. of Athens
Kang Zhang
Tony Yaksh
Varnavas Mouchlis
Matt Buczynski
John Burke
Yasu Kihara
Virgil Woods
Eduard Sabidó
Ruedi Aebersold
Alex Andreyev
Howard Hsu Oswald Quehenberger
Darren Dumlao
Ann Gregus
Jian Cao
Paul Norris
Chris Glass
Eoin Fahy
S. Subramaniam
Andy McCammon
Aaron Armando
Ishita Shah
The LIPID MAPS Consortium
L-R: C.R.H. Raetz, J.L. Witztum, N. Winograd, A.H. Merrill, R.C. Murphy, M.S. VanNieuwenhze,
E.A. Dennis, D.W. Russell, W.A. Shaw, C.K. Glass, J. Chin (NIH), S. Subramaniam, H.A. Brown