1. A story about de novo lipogenesis -more than just a shunting yard for glucose TJ1

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

3. 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?
?????…

4. 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

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

6. 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.

7. 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

8. 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.

9. Acetyl-CoA related pathways

11. More…?

12. 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.

13. 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

14. 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

15. 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

16. 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…

17. Lipogenesis
 Mounier,C., et al., Int J Oncol, (2): p

19. 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.

20. 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

22. Lead optimization to ND-630

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

25. Lead optimization to ND-646

26. 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???

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