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
PERCEPTION OF THE LIPIDOME AFTER THE FIRST DECADE OF LIPIDOMICS
Lipidomics Joins the Omics Evolution (2009), Dennis, PNAS, 106, 2089.
4 base side chains
20 amino acid side chains
nucleic acids, amino acids, sugars and fats: >105 ?
all the fats: >105 ?
Increase in Omics Citations 1985-2009
Wenk MR (2010) Lipidomics: New Tools and Applications. Cell, 143, 888-895. Year
Num
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Lipidomics Publications 2002-2013
Pub
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ions
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LIPID MAPS publications Lipidomics publications
probe
medium cells tissues
homogenate
sonicate
internal standards
extraction
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GC LC
3. Additives (for ionization) 4. Mass spectrometer monitoring modes
(variables) 1. Mass spectrometer types 2. Ionization mode
mass spectrometer
“CLASS”: Comprehensive Lipidomics Analysis using Separation Simplification
category specific (deuterated, odd-chain carbon)
category optimized (liq-liq, SPE)
category specific (GC, NP-HPLC, RP-HPLC, chiral, specialty)
a “divide-and-conquer” strategy
Harkewicz & Dennis, “Applications of mass spectrometry to lipids and membranes” Ann Rev Biochem, 80: 301-325 (2011)
Nuclei – DAPI Mitochondria – MitoTracker Red
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
O-Specific chain
Core
Lipid A
Glycophospholipid
Polysaccharide
Kdo
5-LOX
12-LOX
CYP(EET) 15-LOX
COX CYP(ω)
Cellular eicosanoid metabolism
Buczynski, Dumlao, Dennis, 2009, JLR, 50: 1015-38
Directed Proteomics on Enzymes of Eicosanoid Biosynthesis
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Sabido, Quehenberger Dennis, Aebersold (2012) Mol Cell Proteomics 11: M111.014746
Quehenberger & Dennis, New Eng. J. Medicine, 365, 1812-23 (2011)
Eicosanoid Genes to Proteins to Metabolites Proteins
The Beneficial Lipids in Fish Oil • Fish oil is composed primarily of two ω-‐3 PUFAs:
• 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
Eicosapentaenoic Acid (EPA) (C20:5, ω-‐3)
Docosahexaenoic Acid (DHA) (C22:6, ω-‐3)
ω-‐3
ω-‐3
ω
ω
Arachidonic Acid Serves as a Precursor for Many Eicosanoids
Are eicosanoids less pro-inflammatory when derived from EPA/DHA?
EPA/DHA ?
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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!
Glycerolipids
Fatty Acyls
Glycerophospholipids
Sphingolipids
Sterol Lipids
Prenols
Human Plasma Lipid Categories (µM)
Plasma Lipids in the Metabolic Syndrome
Quehenberger & Dennis, New Eng. J. Medicine, 365, 1812-23 (2011)
Alex Andreyev Howard Hsu Oswald Quehenberger Paul Norris
Darren Dumlao Ann Gregus Jian Cao Aaron Armando Ishita Shah
Varnavas Mouchlis Matt Buczynski John Burke Yasu Kihara
Eoin Fahy S. Subramaniam
Charles Brown Univ. of Missouri
George Kokotos Univ. of Athens
Tony Yaksh
Chris Glass
Kang Zhang
Eduard Sabidó
Ruedi Aebersold
Andy McCammon
Virgil Woods