<p>During oncogenesis, cancer cells go through metabolic reprogramming to maintain their high growth rates and adapt to changes in the microenvironment and the lack of essential nutrients. Several types of cancer are dependent on de novo fatty acid synthesis to sustain their growth rates by providing precursors to construct membranes and produce vital signaling lipids. Fatty acid synthase (FASN) catalyze the terminal step of de novo fatty acid synthesis and it is highly expressed in many types of cancers where it’s up-regulation is correlated with cancer aggressiveness and low therapeutic outcome. Many FASN inhibitors were developed and showed potent anticancer activity however, only one inhibitor advanced to early stage clinical trials with some dose limiting toxicities. Using a modified fluorescence-linked enzyme chemoproteomic strategy (FLECS) screen, we identified HS-106, a thiophenopyrimiden FASN inhibitor that has anti-neoplastic activity against breast cancer in vitro and in vivo. HS-106 was able to inhibit both; purified human FASN activity and cellular fatty acid synthesis activity as evaluated by radioactive tracers incorporation into lipids experiments. In proliferation and apoptosis assays, HS-106 was able to block proliferation and induce apoptosis in several breast cancer cell lines. Several rescue experiment and global lipidome analysis were performed to probe the mechanism by which HS-106 induces apoptosis. HS-106 was found to induce several changes in lipids metabolism: (i) inhibit fatty acids synthesis. (ii) Inhibit fatty acids oxidation as indicated by the ability of inhibiting Malonyl CoA accumulation to block HS-106 induced apoptosis and the increase in the abundance of ceramides. (iii) Increase fatty acids uptake and neutral lipids formation as confirmed 14C Palmitate uptake assay and neutral lipids staining. (iv)Inhibit the formation of phospholipids by inhibiting de novo fatty acid synthesis and diverting exogenous fatty acids to neutral lipids. All of these events would lead to disruption in membranes structure and function. HS-106 was also tested in Lapatinib resistant cell lines and it was able to induce apoptosis and synergizes Lapatinib activity in these cell lines. This may be due the disruption of lipid rafts based on the observation that HS-106 reduces the expression of both HER2 and HER3. HS-106 was found to be well tolerated and bioavailable in mice with high elimination rate. HS-106 efficacy was tested in MMTV neu mouse model. Although did not significantly reduced tumor size (alone), HS-106 was able to double the median survival of the mice and showed potent antitumor activity when combined with Carboplatin. Similar results were obtained when same combinations and dosing schedule was used in C3Tag mouse model except for the inability of HS-106 affect mice survival.</p><p>From the above, HS-106 represent a novel FASN inhibitor that has anticancer activity both in vivo and in vitro. Being a chemically tractable molecule, the synthetic route to HS-106 is readily adaptable for the preparation of analogs that are similar in structure, suggesting that, the pharmacological properties of HS-106 can be improved.</p> / Dissertation

No description available.

Little research has focused on the relationship between lecithin:cholesterol acyltransferase (LCAT) activity and cholesterol metabolism in dogs and cats. To study weight loss and cholesterol metabolism in dogs, four experimental weight-loss diets were fed to 12 obese female beagles for 8 wk in a partial crossover design (n = 6). High- (HGI) or low-glycemic index (LGI) starch and diacylglycerol or triacylglycerol oil were combined to compose diets with similar fatty acid (FA) profiles. Body weight was measured weekly. Fasted blood samples were drawn at wk1, wk4, and wk8 to measure plasma total (TC), unesterified (UC), and esterified cholesterol (EC) concentrations, LCAT activity, and FA composition of the phospholipid (PL) and EC fractions. All groups lost weight. UC increased from wk1 to wk4 (p < 0.05). LCAT activity increased from wk1 to wk4 and remained elevated at wk8 (p < 0.05). Plasma PL FA profiles reflected the diets fed with few diet or time effects. Plasma EC FA profiles reflected the specificity of LCAT for linoleic acid (LA) with minimal diet or time effects. We conclude that weight reduction in dogs occurs in conjunction with increased LCAT activity and altered plasma cholesterol fractions but not changes in plasma PL or EC FA profiles. To measure the activity and demonstrate the FA specificity of LCAT in felines fed varying types of fat, 29 female cats were fed diets enriched with high-oleic sunflower (n = 9), menhaden fish (n = 10), or safflower (n = 10) oil (8g oil/100g kibble) for 4 wk. Fasted blood samples were drawn at d0, d14, and d28 for determination of the blood parameters mentioned previously. LCAT and TC showed no time or diet effects. UC decreased at d28 compared to d0 and d14, while EC increased at d28 compared to d0 and d14 (all p < 0.05). Plasma EC FA profiles reflected the specificity of LCAT for LA with many diet and time effects but contained no docosahexanoic acid (DHA). We conclude that feline LCAT has no measurable affinity for DHA, but both feline and canine LCAT demonstrated specificity for LA regardless of diet fed.

Sixteen horses (2 to 4 yr; 357 to 439 kg BW) were utilized in a randomized complete block design for a 140 d trial to determine effect of omega 3 PUFAs (n-3) supplementation on markers of inflammation in young horses in training. Horses were fed treatments consisting of a control diet (n = 8) fed at 1% BW (as fed) or a treatment diet (n = 8) of concentrate fed at 0.75% BW (as fed) and 350 g of a marine n-3 supplement formulated to provide 15 g of eicosapentaenoic acid (EPA) and 20 g of docosahexaenoic acid (DHA). Body weight and body condition scores (BCS) were obtained biweekly and concentrate adjusted accordingly. Horses were exercised 5 d/wk by students in an equine training course. Type of activity and duration was monitored, along with heart rate to quantify workload. Exercise protocol was divided into 2 phases: phase I (d 0 to110) consisted of ground work and early training under saddle, and phase II (d 111 to 140) consisted of advance maneuvers and moderate workload. Synovial fluid was obtained from right radial carpal joint by arthrocentesis every 28 d and was analyzed for white blood cell count (WBC), total protein (TP), and specific gravity (SG). Serum concentrations of carboxypeptide type II collagen (CPII) and chondroitin sulfate 846 (CS-846) were analyzed by ELISA kits. Dietary treatment did not affect synovial WBC, TP, or SG. Also, concentrations of WBC and TP also did not differ over time. SG increased over time (P < 0.001) as horses moved from phase I to phase II of the trial. Dietary treatment did not influence concentrations of CPII or CS-846. CS-846 tended to increase over time (P = 0.09) and CPII concentrations also increased (P < 0.001) in response to changes in exercise. Furthermore, all horses gained BW and BCS throughout the trial (P < 0.001), but values were not influenced by treatment. This data indicates further studies are needed to determine the efficacy of n-3 supplementation as a preventative measure against development of osteoarthritis.

The effects of partial or total replacement of fish oil with linseed oil or corn oil or both in diets of cobia were valuated. Basal diet was isonitrogenous and isoenergetic and contained 15% crude lipid. Results of the 8-wk feeding trial show that fish fed diet containing only fish oil grew significant better than fish fed other replacement diets (replacement level 33-100%). Fish fed diet containing only plant oil grew the least and had the lowest liver weight, condition factor and body lipid concentration. Oil replacement did not significantly change liver mRNA gene expression of fatty acid desaturase and elongase. As levels of replacement increased, tissue PUFA increased while HUFA decreased. Fish fed all fish oil diet had the highest respiratory burst activities of head kidney phagocytes. Serum of the fish fed the all vegetable oil diets had the lowest lysozyme activities. Fish fed all linseed oil diet had the highest SOD activities. Serum alternative complement pathway activity, aspartate transaminase and alanine transaminase activity did not vary among treatments. The results show that cobia juveniles had relatively high need for fish oil in their diets, and the ability to synthesized HUFA from PUFA was limited. Partial or total replacement (33-100%) of fish oil with linseed oil or corn oil or both were detrimental to fish growth and immune responses.

This research investigated the changes in body adipocyte size and density as well as parameters relating to adipogenesis in growing cobia, Rachycentron canadum. The cobia from a same batch were sampled 5 , 7 , 9 , 13 , 17 , 21 , 26 and 31 weeks after hatching, Liver activities of fatty acid synthase, (FAS), mailc enzyme (ME), and glucose-6-phosphate dehydrogenase (G6PDH) as well as serum total triacylglycerol (TG), total cholesterol (TC), phospholipid (PL), non-esterified fatty acid (NEFA) were assayed. Adipocyte abundance and size spectrum in liver, ventral muscle and dorsal muscle were determined. The cobia attained an average weight of 13.6 ¡Ó 0.5 g at week 5, and were cultured in nursery outdoor ponds until an average weight 87.5 ¡Ó 3.8 g when moved to open ocean cages. They grew to an average weight 1690.8 ¡Ó 106.6 g at week 31. The crude lipid content in liver, ventral and dorsal muscle increased significantly with fish age, and were 36.1 ¡Ó 3.2 g, 14.8 ¡Ó 0.2g, 4.6 ¡Ó 0.0 g at week 31. No significant change was found in hepatic FAS activity. Hepatic ME activity increased significantly after the fish were stocked in the cages. Whilr hepatic ME activity reduced with age, TG and PL were both increased significantly with age. While adipocyte diameter in liver showed a decreasing trend as the fish grew its abundance increased significantly at week 13 and maintained between 3070¡ã3356 x 104 cell/g tissue during week 17-31. Similarly, adipocyte abundance in ventral muscle increased significantly at week 13, and maintained between 255¡ã269 x 104 cell/g tissue afterwards. There was no significant change in size of ventral muscle adipocyte as fish grew. These results indicate that the adipocyte hyperplasia occurred during week 7- 13, when body weight was 100- 400 g. Adipocyte hypertrophy was observed when the fish reached week 31, there was a sign of adipocyte hypertrophy in liver when the fish were at the last phase of nursery outdoor ponds before being moved to the offshore cages.

Heart-type fatty acid binding protein (H-FABP) is a major fatty acid binding factor in skeletal muscles. Genetic lack of H-FABP severely impairs the esterification and oxidation of exogenous fatty acids in soleus muscles isolated from chow-fed mice (CHOW-solei) and high fat diet-fed mice (HFD-solei), and prevents the HFD-induced accumulation of muscle triglycerides. Here, we examined the impact of H-FABP deficiency on the relationship between fatty acid utilization and glucose oxidation. Glucose oxidation was measured in isolated soleus muscles in the presence or absence of 1 mM palmitate (simple protocol) or in the absence of fatty acid after preincubation with 1 mM palmitate (complex protocol). With the simple protocol, the mutation slightly reduced glucose oxidation in CHOW-muscles, but markedly increased it in HFDmuscles; unexpectedly, this pattern was not altered by the addition of palmitate, which reduced glucose oxidation in both CHOW- and HFD-solei irrespective of the mutation. In the complex protocol, the mutation first inhibited the synthesis and accumulation of triglycerides and then their mobilization; with this protocol, the mutation increased glucose oxidation in both CHOW- and HFD-solei. We conclude: (i) H-FABP mediates a non-acute inhibition of muscle glucose oxidation by fatty acids, likely by enabling both the accumulation and mobilidoes not mediate the acute inhibitory effect of extracellular fatty acids on muscle glucose oxidation; (iii) H-FABP affects muscle glucose oxidation in opposing ways, with inhibition prevailing at high muscle triglyceride contents.zation of a critical mass of muscle triglycerides; (ii) H-FABP

This research studied the dietary effect of supplemental plant oils on growth, adiposity and lipid metabolism-related enzyme activity of juvenile cobia. The isonitrogenous and isoenergetic basal diet contained 15% crude lipid, 6% fish oil and 9% supplemental oils. The supplemental oils were varied among 5 dietary treatments, including fish oil (HUFA, n-3)(FO), perilla oil (18:3 n-3)(PE), safflower oil (18:2 n-6)(SA), olive oil (18:1 n-9)(OL), and palm oil (16:0)(PA). Results of the 10-wk feeding trial show that fish fed diet containing palm oil had the highest final weight, and was significant higher than fish fed SA diet. SA group had highest crude lipid concentration. OL group had the least crude protein concentration. PA group had the highest ash concentration. FO group had the highest crude protein and moisture content, the least crude lipid and ash content. Adipocyte density in various, tissues did not vary with time, except the ventral fat depot. Tissue adipocyte density of FO group was the least. Adipocyte density of PE group was higher than FO group, and its mean adipcoyte diameter in dorsal muscle was great than the other groups. Fatty acid synthase (FAS), as measured by specific activity, decreased with times in the PE group. Adipocyte density of SA group the highest of all groups, and its mean adipocyte diameter in dorsal muscle was also the greatest. Adipocyte density in the ventral fat depot of OL group was the highest, and its mean diameter in dorsal muscle was the smallest. Adipocyte density of PA group was only slightly lower than SA group. The tissue acid composition of the cobia was influenced by the supplemental plant oils. Tissue HUFA concentration and n-3/n-6 ratio was decreased, MUFA, PUFA and SAF composition was increased when the plant oils were supplemented. The results show that the supplementation of the plant oils could affect the density , size and tissue distribution of adipocytes, fatty acid synthesis pathway in the liver and tissue fatty acid composition. Feeding the cobia diet containing supplemental safflower or palm oil significantly increased density and cell size of adipocytes in the tissue of the cobia.

The present study cloned successfully two lipid-metabolism genes, lipoprotein lipase (LPL) and fatty acid binding protein (FABPs) from cobia and studied the mRNA expressions of the two genes and their upstream gene PPARs when the cobia were fed diets containing 15% lipid. Among the lipids, 6% was fish oil and the remaining 9% were supplemented by fish oil (FO, rich in n-3 HUFA), perilla oil (PE, rich in 18:2 n-6), safflower oil (SA, rich in 18:2 n-6), olive oil (OL, rich in 18:1 n-9) or palm oil (PA, rich in 16:0). The whole sequences of LPL, liver-FABP (L-FABP) and muscle-FABP (M-FABP) encode 520, 126 and 133 amino acids, respectively. RT-PCR and real time PCR analyses based on these gene sequences show that the mRNA expressions of L-FABP and M-FABP in the tissue of the cobia were diet-specific. The mRNA expression of LPL, on the other hand, did not respond to the treatments, except in visceral fat depot. Linear regression analysis shows that the mRNA expression of LPL in the liver and muscle was positively (P<0.05) related to dietary fatty acids and ther concentration, but that in the visceral fat depot was negatively related. The mRNA expression of FABPs was also positively correlated with dietary fatty acid levels. Among all fatty acids, the levels of C14:0, C20:1 n-9, EPA and DHA were positively correlated with the mRNA expression of PPAR£^and also with FABPs mRNA expression in the visceral fat depot and LPL mRNA expression in the muscle. Thus, LPL, L-FABP and M-FABP mRNA expression of the cobia were highly influenced by the kind and amount of dietary fatty acids. The role of PPARs was not clearly demonstrated.

Thesis (Ph. D.)--Rutgers University, 2009. / "Graduate Program in Nutritional Sciences." Includes bibliographical references (p. 149-161).