Search

Human FASN is the key enzyme required for de novo synthesis of fatty acids. Up-regulated FASN expression has been reported in various human cancers and was thought to contribute to poor prognosis and recurrence of these cancers. Studies using model cell lines have indicated the role of FASN in both intrinsic and acquired drug and radiation resistance. Recent studies suggest that FASN may play an important role in regulating gene expression such as pro-apoptotic proteins and cellular processes such as DNA repair pathways, which in turn contribute to resistance to drug and radiation-induced apoptosis. In this review, we will highlight our recent progress in understanding the mechanism of FASN-induced resistance.

Fatty acid synthase (FASN), the sole cytosolic mammalian enzyme for de novo lipid synthesis, is crucial for cancer cell survival and associates with poor prognosis. FASN overexpression has been found to cause resistance to genotoxic insults. Here we tested the hypothesis that FASN regulates DNA repair to facilitate survival against genotoxic insults and found that FASN suppresses NF-κB but increases specificity protein 1 (SP1) expression. NF-κB and SP1 bind to a composite element in the poly(ADP-ribose) polymerase 1 (PARP-1) promoter in a mutually exclusive manner and regulate PARP-1 expression. Up-regulation of PARP-1 by FASN in turn increases Ku protein recruitment and DNA repair. Furthermore, lipid deprivation suppresses SP1 expression, which is able to be rescued by palmitate supplementation. However, lipid deprivation or palmitate supplementation has no effect on NF-κB expression. Thus, FASN may regulate NF-κB and SP1 expression using different mechanisms. Altogether, we conclude that FASN regulates cellular response against genotoxic insults by up-regulating PARP-1 and DNA repair via NF-κB and SP1.

Fatty acid synthase (FASN), the enzyme responsible for de novo synthesis of free fatty acids, is up-regulated in many cancers. FASN is essential for cancer cell survival and contributes to drug resistance and poor prognosis. However, it is not expressed in most nonlipogenic normal tissues. Thus, FASN is a desirable target for drug discovery. Although different FASN inhibitors have been identified, none has successfully moved into clinical use. In this study, using in silico screening of an FDA-approved drug database, we identified proton pump inhibitors (PPIs) as effective inhibitors of the thioesterase activity of human FASN. Further investigation showed that PPIs inhibited proliferation and induced apoptosis of cancer cells. Supplementation of palmitate, the end product of FASN catalysis, rescued cancer cells from PPI-induced cell death. These findings provide new evidence for the mechanism by which this FDA-approved class of compounds may be acting on cancer cells.

Fatty acid synthase (FASN), the sole protein capable of de novo synthesis of free fatty acids, is overexpressed in a wide variety of human cancers and is associated with poor prognosis and aggressiveness of these cancers. Orlistat, an FDA-approved drug for obesity treatment that inhibits pancreatic lipases in the GI tract, also inhibits the thioesterase (TE) of human FASN. The cocrystal structure of TE with orlistat shows a pseudo TE dimer containing two different forms of orlistat in the active site, an intermediate that is covalently bound to a serine residue (Ser2308) and a hydrolyzed and inactivated product. In this study, we attempted to understand the mechanism of TE-catalyzed orlistat hydrolysis by examining the role of the hexyl tail of the covalently bound orlistat in water activation for hydrolysis using molecular dynamics simulations. We found that the hexyl tail of the covalently bound orlistat undergoes a conformational transition, which is accompanied by destabilization of a hydrogen bond between a hydroxyl moiety of orlistat and the catalytic His2481 of TE that in turn leads to an increased hydrogen bonding between water molecules and His2481 and increased chance for water activation to hydrolyze the covalent bond between orlistat and Ser2308. Thus, the conformation of the hexyl tail of orlistat plays an important role in orlistat hydrolysis. Strategies that stabilize the hexyl tail may lead to the design of more potent irreversible inhibitors that target FASN and block TE activity with greater endurance.

Fatty acid synthase (FASN), a seven domain enzyme, is the sole protein capable of de novo synthesis of free fatty acids, most commonly 16-carbon palmitate. The fatty acid synthesis cycle begins with the condensation of acetyl-CoA and malonyl-CoA, and continues with the elongation of the fatty acid chain, which is tethered to an acyl carrier protein domain (ACP), via a repeating cycle. At the end of elongation, the thioesterase (TE) domain of FASN, a member of the serine hydrolase family with an Asp-His-Ser catalytic triad, cleaves the bond between palmitate and ACP, and releases free palmitate. FASN has been found to be over-expressed in a wide variety of human cancers, and this over-expression is correlated to a higher metastatic potential and poorer prognosis in cancer patients. FASN over-expression is also associated with increased resistance toward cancer chemotherapeutics. Orlistat, an FDA approved drug for obesity treatment that prevents uptake of dietary fats by inhibiting pancreatic lipases in the gastrointestinal tract, is a compound found to act as a reversible inhibitor of FASN TE. A previous study using the co-crystal structure of the TE domain with orlistat found that orlistat was covalently bound to the active site Ser within the TE domain, indicating that inhibition of FASN by orlistat halts the fatty acid synthesis cycle by blocking the release of free palmitate from the ACP. A hydrolyzed form of orlistat was also observed in the active site of TE, demonstrating that orlistat is not a stable inhibitor of FASN. In this study, we examined the mechanism of orlistat hydrolysis within the TE domain of FASN using molecular dynamics simulations. We found that the hexanoyl tail of orlistat is capable of shifting while covalently bound to the active site Ser, and that this shift is accompanied by the destabilization of a hydrogen bond that exists between a hydroxyl moiety of orlistat and the active site His. Once this hydrogen bond is destabilized, a catalytic water molecule can enter the active site of TE with the proper orientation for catalysis of the covalent bond between orlistat and Ser. These findings suggest that the hexanoyl tail of orlistat plays an important role in its hydrolysis and may guide the future design of new inhibitors that target the TE domain of FASN with greater endurance for potential use in the treatment of cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3978. doi:1538-7445.AM2012-3978