Specific classes of long chain fatty acids have been implicating in promoting various diseases including obesity, metabolic syndrome, diabetes, and cardiovascular disease. Additionally, there is a strong correlation between dietary intake of saturated and trans-unsaturated fatty acids with certain cancers including especially, colorectal and breast. The focus of the present work is to devise high throughput methods to identify compounds that selectively inhibit fatty acid uptake into cells. Fatty acid uptake is a protein-mediated event requiring a fatty acid transport protein (FATP) and an acyl-CoA synthetase (ACSL). Our laboratory has identified and extensively characterized yeast strains deficient in this uptake system. The transport defect of yeast is complemented by mammalian orthologs of the yeast genes and the mammalian transport system reconstituted in yeast is the target for the proposed automated inhibitor screening method development.
The specific aims of the proposal are to:  demonstrate the sensitivity and specificity of the mammalian transport proteins expressed in the yeast live cell system for analysis in a 96-well high throughput format;  to conduct a proof or principle screen of a diversified small molecule library for compounds that inhibit fatty acid uptake; and  to devise secondary screens which employ human cells in culture. Our ultimate goal is to identify compounds that inhibit fatty acid uptake into cells without cytotoxic effects. Such compounds are expected to be useful to prevent intestinal absorption of dietary lipid and/or uptake into other cell types to prevent cell damage caused by excessive concentrations of fatty acids, which lead to disease.
|Black, Paul N; Ahowesso, Constance; Montefusco, David et al. (2016) Fatty Acid Transport Proteins: Targeting FATP2 as a Gatekeeper Involved in the Transport of Exogenous Fatty Acids. Medchemcomm 7:612-622|
|Saini, Nipun; Black, Paul N; Montefusco, David et al. (2015) Fatty acid transport protein-2 inhibitor Grassofermata/CB5 protects cells against lipid accumulation and toxicity. Biochem Biophys Res Commun 465:534-41|
|Ahowesso, Constance; Black, Paul N; Saini, Nipun et al. (2015) Chemical inhibition of fatty acid absorption and cellular uptake limits lipotoxic cell death. Biochem Pharmacol 98:167-81|
|Sandoval, Angel; Chokshi, Aalap; Jesch, Elliot D et al. (2010) Identification and characterization of small compound inhibitors of human FATP2. Biochem Pharmacol 79:990-9|
|Black, Paul N; Sandoval, Angel; Arias-Barrau, Elsa et al. (2009) Targeting the fatty acid transport proteins (FATP) to understand the mechanisms linking fatty acid transport to metabolism. Immunol Endocr Metab Agents Med Chem 9:11-17|
|Li, Hong; Black, Paul N; Chokshi, Aalap et al. (2008) High-throughput screening for fatty acid uptake inhibitors in humanized yeast identifies atypical antipsychotic drugs that cause dyslipidemias. J Lipid Res 49:230-44|