title: Chemical inhibition of fatty acid absorption and cellular uptake limits lipotoxic cell death.
abstract: Chronic elevation of plasma free fatty acid (FFA) levels is commonly associated with obesity, type 2 diabetes, cardiovascular disease and some cancers. Experimental evidence indicates FFA and their metabolites contribute to disease development through lipotoxicity. Previously, we identified a specific fatty acid transport inhibitor CB16.2, a.k.a. Lipofermata, using high throughput screening methods. In this study, efficacy of transport inhibition was measured in four cell lines that are models for myocytes (mmC2C12), pancreatic β-cells (rnINS-1E), intestinal epithelial cells (hsCaco-2), and hepatocytes (hsHepG2), as well as primary human adipocytes. The compound was effective in inhibiting uptake with IC50s between 3 and 6μM for all cell lines except human adipocytes (39μM). Inhibition was specific for long and very long chain fatty acids but had no effect on medium chain fatty acids (C6-C10), which are transported by passive diffusion. Derivatives of Lipofermata were evaluated to understand structural contributions to activity. Lipofermata prevented palmitate-mediated oxidative stress, induction of BiP and CHOP, and cell death in a dose-dependent manner in hsHepG2 and rnINS-1E cells, suggesting it will prevent induction of fatty acid-mediated cell death pathways and lipotoxic disease by channeling excess fatty acids to adipose tissue and away from liver and pancreas. Importantly, mice dosed orally with Lipofermata were not able to absorb (13)C-oleate demonstrating utility as an inhibitor of fatty acid absorption from the gut.
authors.affiliation: Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0664, United States.
authors.forename: Constance
authors.initials: C
authors.lastname: Ahowesso
authors.affiliation: Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0664, United States.
authors.forename: Paul N
authors.initials: PN
authors.lastname: Black
authors.affiliation: Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0664, United States.
authors.forename: Nipun
authors.initials: N
authors.lastname: Saini
authors.affiliation: Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0664, United States.
authors.forename: David
authors.initials: D
authors.lastname: Montefusco
authors.affiliation: Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0664, United States.
authors.forename: Jessica
authors.initials: J
authors.lastname: Chekal
authors.affiliation: Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232, United States; Vanderbilt Specialized Chemistry Center for Probe Development (MLPCN), Nashville, TN 37232, United States.
authors.forename: Chrysa
authors.initials: C
authors.lastname: Malosh
authors.affiliation: Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232, United States; Vanderbilt Specialized Chemistry Center for Probe Development (MLPCN), Nashville, TN 37232, United States; Department of Chemistry, Vanderbilt University, Nashville, TN 37232, United States.
authors.forename: Craig W
authors.initials: CW
authors.lastname: Lindsley
authors.affiliation: Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232, United States; Vanderbilt Specialized Chemistry Center for Probe Development (MLPCN), Nashville, TN 37232, United States; Department of Chemistry, Vanderbilt University, Nashville, TN 37232, United States.
authors.forename: Shaun R
authors.initials: SR
authors.lastname: Stauffer
authors.affiliation: Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0664, United States. Electronic address: cdirusso2@unl.edu.
authors.forename: Concetta C
authors.initials: CC
authors.lastname: DiRusso
mesh_terms: D017667:Adipocytes / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D045744:Cell Line, Tumor; D005227:Fatty Acids / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D015394:Molecular Structure; D054852:Small Molecule Libraries; D013141:Spiro Compounds / Q000494:pharmacology*; D013830:Thiadiazoles / Q000494:pharmacology*
chemical_list: C000604698:5'-bromo-5-phenyl-spiro(3H-1,3,4-thiadiazole-2,3'-indoline)-2'-one; D005227:Fatty Acids; D054852:Small Molecule Libraries; D013141:Spiro Compounds; D013830:Thiadiazoles
keywords: 5′-bromo-5-phenyl-spiro[3H-1,3,4-thiadiazole-2,3′-indoline]-2′-one; FATP2 inhibitor; Fatty acid transport; Lipid droplet; Lipotoxicity