Many lipids and their derivatives are potent ligands for cellular receptors. The action of lipid second messengers is regulated by lipases and related enzymes that limit the lifetime and sphere of action of these bioactive molecules. The products of these hydrolytic reactions can themselves be second messengers and precursors to other bioactive molecules. For example, the serine hydrolase monoacylglycerol lipase (MAGL) is responsible for limiting the lifetime and sphere of action of the endocannabinoid 2-arachidonoylglycerol (2-AG) and for generating the fatty acid product, arachidonic acid. Inhibitors of these enzymes are potential therapeutics for the treatment of pain, anxiety, addiction, inflammation, and a myriad of diseases where inflammation or lipid regulation play central roles. However, the tools currently available for the evaluation of serine hydrolase activity and inhibition are primarily limited to use in vitro. In this grant, we will develop luminogenic sensors for serine hydrolases (Aim 1), and apply them to the imaging of serine hydrolase activity in live cells and identification of novel inhibitors for this class of enzymes (im 2). Finally, we will perform real-time imaging of serine hydrolase activity in vivo, to determine where and when these enzymes are active, and how the action of drug pumps affect the tissue distribution of small molecule inhibitors of their function (Aim 3).
Many signaling molecules in the body are derived from fats and play prominent roles in the perception of pain, inflammation, and the progression of a myriad of diseases. The goal of this research is to create light-emitting sensors of the enzymes that regulate these lipid signaling molecules. Our sensors will literally illuminate the cells and tissues where these enzymes are active, helping us to study these enzymes and to develop drugs to treat conditions resulting from their misregulation.
| Miller, Stephen C; Mofford, David M; Adams Jr, Spencer T (2018) Lessons Learned from Luminous Luciferins and Latent Luciferases. ACS Chem Biol 13:1734-1740 |
| Sharma, Deepak K; Adams Jr, Spencer T; Liebmann, Kate L et al. (2017) Rapid Access to a Broad Range of 6'-Substituted Firefly Luciferin Analogues Reveals Surprising Emitters and Inhibitors. Org Lett 19:5836-5839 |
| Adams Jr, Spencer T; Mofford, David M; Reddy, G S Kiran Kumar et al. (2016) Firefly Luciferase Mutants Allow Substrate-Selective Bioluminescence Imaging in the Mouse Brain. Angew Chem Int Ed Engl 55:4943-6 |
| Mofford, David M; Miller, Stephen C (2015) Luciferins behave like drugs. ACS Chem Neurosci 6:1273-5 |
| Mofford, David M; Adams Jr, Spencer T; Reddy, G S Kiran Kumar et al. (2015) Luciferin Amides Enable in Vivo Bioluminescence Detection of Endogenous Fatty Acid Amide Hydrolase Activity. J Am Chem Soc 137:8684-7 |
