title: Development of a novel, cell-based chemical screen to identify inhibitors of intraphagosomal lipolysis in macrophages.
abstract: Macrophages play a central role in tissue homeostasis and the immune system. Their primary function is to internalize cellular debris and microorganisms for degradation within their phagosomes. In this context, their capacity to process and sequester lipids such as triacylglycerides and cholesteryl esters makes them key players in circulatory diseases, such as atheroclerosis. To discover new inhibitors of lipolytic processing within the phagosomal system of the macrophage, we have developed a novel, cell-based assay suitable for high-throughput screening. We employed particles carrying a fluorogenic triglyceride substrate and a calibration fluor to screen for inhibitors of phagosomal lipolysis. A panel of secondary assays were employed to discriminate between lipase inhibitors and compounds that perturbed general phagosomal trafficking events. This process enabled us to identify a new structural class of pyrazole-methanone compounds that directly inhibit lysosomal and lipoprotein lipase activity.
authors.affiliation: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Brian C
authors.initials: BC
authors.lastname: VanderVen
authors.forename: Albin
authors.initials: A
authors.lastname: Hermetter
authors.forename: Amy
authors.initials: A
authors.lastname: Huang
authors.forename: Fredrick R
authors.initials: FR
authors.lastname: Maxfield
authors.forename: David G
authors.initials: DG
authors.lastname: Russell
authors.forename: Robin M
authors.initials: RM
authors.lastname: Yates
mesh_terms: D000818:Animals; D005453:Fluorescence; D057166:High-Throughput Screening Assays / Q000379:methods*; D008066:Lipolysis / Q000187:drug effects*; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D010588:Phagosomes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D011721:Pyrenes / Q000378:metabolism; D054852:Small Molecule Libraries / Q000032:analysis* / Q000494:pharmacology*; D002787:Sterol Esterase / Q000037:antagonists & inhibitors / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism
chemical_list: D011721:Pyrenes; D054852:Small Molecule Libraries; D014280:Triglycerides; C030984:pyrene; D002787:Sterol Esterase; D008071:Lipoprotein Lipase