The overall goal of this K99/R00 career development award proposal is to integrate innovate chemical probes, including in vivo-active small-molecule inhibitors and specialized chemoproteomic assays, with well- established genetic knockout and behavioral mouse models to obtain a global view of the pathophysiological consequence of disrupting 2-arachidonoylglycerol (2-AG) biosynthesis in mouse models of obesity and inflammation. The endogenous cannabinoid (endocannabinoid) 2-AG is a lipid transmitter that activates the G- protein-coupled receptors CB1 and CB2, which are also targets for the psychoactive ingredient in marijuana, 9-tetrahydrocannabinol. The importance of the endocannabinoid system in obesity-associated metabolic disorders is highlighted by the clinical activity of CB1 antagonists as anti-obesity and anti-diabetic drugs. Complementary studies in preclinical models have since shown that many of the beneficial effects are due to blocking CB1 receptors at peripheral sites. These studies highlight the need for a deeper understanding of the central and/or peripheral contributions of the endocannabinoid system in regulating energy homeostasis as well as obesity-related disease states. 2-AG biosynthesis in vivo is differentially regulated by two sequence- related enzymes, diacylglycerol lipase-alpha and beta (DAGL alpha and DAGL beta, respectively), providing an experimental (and, eventually translational) opportunity to uncouple central and peripheral endocannabinoid signaling through selective inactivation of DAGL isoforms. This proposal will test the hypothesis that DAGL alpha and DAGL beta biosynthesize 2-AG involved in energy homeostasis and perform complementary functions in energy balance through regulation of CB1-dependent signaling at anatomically-distinct sites.
The specific aims are to 1) measure energy homeostasis parameters and body mass composition in vivo in DAGL-disrupted mice 2) measure adipose tissue macrophage accumulation in DAGL-disrupted mice and 3) Determine the physiological effects of inactivating DAGLs in the development and progression of metabolic syndrome. Our preliminary studies show that DAGL alpha display a lean phenotype despite increased food intake, providing evidence of altered energy balance upon central and/or peripheral disruption of 2-AG biosynthesis. Our preliminary studies also show the feasibility of using DAGL-selective inhibitors to uncouple central and peripheral 2-AG signaling pathways in vivo. The candidate aims to combine previous training in chemoproteomics, synthetic chemistry, and quantitative proteomics and metabolomics with new mentored training in mouse models of metabolism and inflammation including non-invasive measurements of metabolic parameters governing energy homeostasis and body mass composition as well as in vivo tracking of adipose tissue macrophage accumulation using adoptive cell transfer and flow cytometry.

Public Health Relevance

Obesity and associated metabolic disorders, including type 2 diabetes have been associated with increased signaling activity of the principal endogenous cannabinoid lipid, 2-arachidonoylglycerol (2-AG). The importance of the endogenous cannabinoid system in obesity-associated metabolic disorders is highlighted by the clinical activity of cannabinoid receptor type 1 (CB1) antagonists as anti-obesity and anti-diabetic drugs. The goal of this proposal is to determine the tissue-specific functions of 2-AG biosynthetic pathways in energy balance and test whether blockade of 2-AG production in vivo can produce therapeutic effects akin to systemic CB1 antagonists in mouse models of obesity.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Career Transition Award (K99)
Project #
1K99DA035864-01A1
Application #
8701001
Study Section
Special Emphasis Panel (ZDA1-EXL-T (02))
Program Officer
Sorensen, Roger
Project Start
2014-06-15
Project End
2016-05-31
Budget Start
2014-06-15
Budget End
2015-05-31
Support Year
1
Fiscal Year
2014
Total Cost
$128,250
Indirect Cost
$9,500
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Manna, Joseph D; Wepy, James A; Hsu, Ku-Lung et al. (2014) Identification of the major prostaglandin glycerol ester hydrolase in human cancer cells. J Biol Chem 289:33741-53
Grim, Travis W; Ghosh, Sudeshna; Hsu, Ku-Lung et al. (2014) Combined inhibition of FAAH and COX produces enhanced anti-allodynic effects in mouse neuropathic and inflammatory pain models. Pharmacol Biochem Behav 124:405-11
Inloes, Jordon M; Hsu, Ku-Lung; Dix, Melissa M et al. (2014) The hereditary spastic paraplegia-related enzyme DDHD2 is a principal brain triglyceride lipase. Proc Natl Acad Sci U S A 111:14924-9