A story about de novo lipogenesis -more than just a shunting yard for glucose TJ 1

Sampled data at routine work ACC1 enzymic assay ACC2 enzymic assay C14-acetate incorporation assay

Why more and more Fasyn project coming? What is ACC? How it works? What did we do in the past? Where is Acetate-CoA from? Where to go? Any other targets? How does fatty acid synthesis (FAsyn) induce diseases? What is ACL? What two protocol used for, Chloroform/methanol VS. Saponification? The relationship between FAsyn and cancer? ?????…

Inhibition of ACC1 and ACC2 activity Continuous Fatty Acid Oxidation and Reduced Fat Storage in Mice Lacking Acetyl-CoA Carboxylase 2 Abuelheiga,L., et al., Science, 2001.  Tong, L. and H.J. Harwood, Jr., J Cell Biochem, 2006. Harwood, H. J., Jr., et al. (2003) J Biol Chem

Distinct roles of ACC1 and ACC2 in FA metabolism  Tong, L. and H.J. Harwood, Jr., J Cell Biochem, 2006.

Structures of biotin carboxylase (BC) domain of ACC Harriman, G., et al. (2016). PNAS Tong, L. (2005). Cell Mol Life Sci Svensson, R. U., et al. (2016). Nat Med.

ACC activity and FAsyn regulation Wang, C., et al. (2015). "Expert Rev Anticancer Ther SREBP-1c: regulatory element-binding protein-1c ChREBP: carbohydrate response element-binding protein AMPK: AMP-activated protein kinase LXRs: liver X receptors There are three SREBP family members, designated SREBP-1a, -1c, and -2. The SREBP-1 isoforms predominantly regulate genes involved in FA and TG synthesis while the SREBP-2 isoform activates genes involved in cholesterol biosynthesis and cellular homeostasis Loubiere, C., et al. (2015). Oncotarget ACC activity regulation Phosphorylation/dephosphorylation by AMPK Transcriptional, mediated by SREBP-1c, ChREBP and liver X receptors (LXRs) Allosteric activation by high concentrations of citrate induced polymerization Kim, C. W., et al. (2010). PNAS

ACC inhibitors ND-630 ACC2 selective neurological and cardiovascular safety events ACC1/2 unselective increased lipid oxidation, can’t decrease TG and weight Harriman, G., et al. (2016). PNAS Griffith, D.A., et al., J Med Chem,2014 Svensson, R. U., et al. (2016). Nat Med.

Acetyl-CoA related pathways

More…?

From the very beginning Lipids Glucose The majority of adult mammalian tissues satisfy their lipid requirements through the uptake of free fatty acids (FFAs) and lipoproteins, such as low-density lipoprotein (LDL), from the bloodstream. Fatty acid (FA) and cholesterol biosynthesis are restricted to a subset of tissues, including liver, adipose and lactating breast tissues.

Lipid biosynthesis Baenke, F., et al. (2013). Dis Model Mech Detailed reaction refers to: Sanders, F. W. and J. L. Griffin (2016) Biol Rev Camb Philos Soc

Wnt/LRP6 regulation of LDL uptake and DNL Song, K., et al. (2014) Oncotarget LRP6 forms a complex with LDLR and other proteins of endocytic machinery involved in LDL LRP6 inhibits IGF1-IGF1R dependent activation of AKT/SREBP, DNL and PPARγ-dependent adipogenesis

Fat cell Most normal cells use dietary and/or exogenous lipids for synthesis of new structural lipids . Active FAsyn pathway in adipocytes, hepatocytes, hormone-sensitive cells and fetal lung tissue. In general, de novo fatty acid synthesis is suppressed in most normal cells. Sanders, F. W. and J. L. Griffin (2016). Biol Rev Camb Philos Soc

Fatty acid and cancer Cancer tissues can generate lipids Tumor takes up lipids from tissue environment Medes, G., et al. (1953). Cancer Res Tumor specific antigen OA-519 is FASN Kuhajda, F. P., et al. (1994). PNAS FAsyn inhibitors in different preclinical models Kuhajda, F. P., et al. (2000). PNAS FAsyn inhibition can block cancer growth Abramson, H. N. (2011). J Med Chem Zaidi, N., et al. (2012). Cancer Res THEN, ACC, SCD, ACLY…

Lipogenesis  Mounier,C., et al., Int J Oncol, 2014. 45(2): p. 485-92.

DNL in the control of metabolic homeostasis Solinas, G., et al. (2015). Mol Metab Sanders, F. W. and J. L. Griffin (2016). Biol Rev Camb Philos Soc DNL produces lipids by disposing of glucose and calories. DNL supports the synthesis of several signaling molecules implicated in the control of metabolic homeostasis.

The roles of lipids in the tumor microenvironment Hypoxia means low oxygen HIF, hypoxia inducible factor, a transcription factor mediates cellular response to hypoxia Baenke, F., et al. (2013). Dis Model Mech

Lead optimization to ND-630

Acute in vivo efficacy of ND-630 in rats Chow-fed male Sprague-Dawly rats HSD-induced obesity ZDF rats

Lead optimization to ND-646

Final answer for ACC phosphorylation inhibition Svensson, R. U., et al. (2016). Nat Med. calyculin A (Cal A), protein phosphatase inhibitor To test how ND-646 inhibit p-ACC: Did not prevent the ability of AMPK to phosphorylate hACC2 at Ser222 No effect on other AMPK substrates Might promote dephosphorylation of p-Ser hACC1 R172A mutation, p-Ser was gone; +Cal A, p-Ser restored SO WHAT???

Fat isn’t just something we eat: it may also lie at the heart of a new approach to treating cancer.