Submitted on October 10, 2008
Revised on February 11, 2009
Accepted on February 24, 2009
Uncovering multiple molecular targets for caffeine by a drug target validation strategy of combined A2A receptor knockouts and microarray profiling
Liqun Yu1, Joana Coelho1, Xiaoling Zhang1, Yutao Fu2, Abigail Tillman1, Ulas Karaoz2, Bertil B Fredholm3, Zhiping Weng2, and Jiang-Fan Chen1*
1 Boston University School of Medicine
2 Boston University
3 Karolinska Institute
* To whom correspondence should be addressed. E-mail: chenjf{at}bu.edu.
Caffeine is the most widely consumed psychoactive substance and has complex pharmacological actions in brain. In this study, we employed a novel drug target validation strategy to uncover the multiple molecular targets of caffeine using combined A2A receptor (A2AR) knockouts (KO) and microarray profiling. Caffeine (10 mg/kg) elicited a distinct profile of striatal gene expression in WT mice when compared to that by A2AR gene deletion or by administering caffeine into A2AR KO mice. Thus, A2ARs are required but not sufficient to elicit the striatal gene expression by caffeine (10 mg/kg). Caffeine (50 mg/kg) induced complex expression patterns with three distinct sets of striatal genes: (1) one-subset overlapped with those elicited by genetic deletion of A2ARs; (2) the second subset elicited by caffeine in WT as well as A2AR KO mice; and (3) the third subset elicited by caffeine only in A2AR KO mice. Furthermore, striatal gene sets elicited by the phosphodiesterase (PDE) inhibitor rolipram and the GABAA receptor antagonist bicucullin, overlapped with the distinct subsets of striatal genes elicited by caffeine (50 mg/kg) administered to A2AR KO mice. Finally, Gene Set Enrichment Analysis reveals that adipocyte differentiation/insulin signaling is highly enriched in the striatal gene sets elicited by both low and high doses of caffeine. The identification of these distinct striatal gene populations and their corresponding multiple molecular targets, including A2AR, non-A2AR (possibly A1Rs and pathways associated with PDE and GABAAR) and their interactions, and the cellular pathways affected by low and high doses of caffeine, provides molecular insights into the acute pharmacological effects of caffeine in the brain.
