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Open Full Page Published OnlineFirst July 7, 2015; DOI: 10.1158/1078-0432.CCR-14-3300 Molecular Pathways Clinical Cancer Research Molecular Pathways: Is AMPK a Friend or a Foe in Cancer? D. Grahame Hardie Abstract The AMP-activated protein kinase (AMPK) is a sensor of cellular including drugs in long use such as salicylate and metformin, energy status expressed in essentially all eukaryotic cells. Once and there is evidence that regular use of either of the latter activated by energetic stress via a mechanism that detects increases provides protection against development of cancer. Tumor cells in AMP:ATP and ADP:ATP ratios, AMPK acts to restore energy appear to be under selection pressure to downregulate AMPK, homeostasis by switching on catabolic pathways that generate thus limiting its restraining influenceoncellgrowthand ATP, while switching off ATP-consuming processes, including proliferation, and several interesting mechanisms by which anabolic pathways required for cell growth and proliferation. this occurs are discussed. Paradoxically, however, a complete AMPK activation promotes the glucose-sparing, oxidative metab- loss of AMPK function, which appears to be rare in human olism utilized by most quiescent cells, rather than the rapid cancers, may be deleterious to survival of tumor cells. AMPK glucose uptake and glycolysis used by most proliferating can therefore be either a friend or a foe in cancer, depending cells. Numerous pharmacologic activators of AMPK are known, on the context. Clin Cancer Res; 21(17); 3836–40. Ó2015 AACR. Background cellular energy status by detecting increases in these ratios. In all species, it is activated >100-fold by phosphorylation of a con- The AMP-activated protein kinase (AMPK) is a sensor of cellular served threonine residue (Thr172 in rat a2; ref. 5) located within energy status and a key regulator of energy homeostasis, which the "activation loop" of the a subunit kinase domain. The primary exists universally in eukaryotes as heterotrimeric complexes con- upstream kinase phosphorylating this site in mammalian cells is a taining catalytic a and regulatory b, and g subunits (1, 2). In complex comprising the protein kinase LKB1 and two accessory humans, multiple isoforms of each subunit (AMPK-a1, -a2; -b1, subunits, STRAD and MO25 (6). Heterozygous mutations in -b2; -g1, -g2, -g3) are encoded by distinct genes (PRKAA1, STK11, the human gene encoding LKB1, had been identified as PRKAA2; PRKAB1, PRKAB2; PRKAG1, PRKAG2, PRKAG3), gen- the cause of Peutz–Jeghers syndrome, an inherited susceptibility erating up to 12 heterotrimeric combinations. In the yeast Sac- to cancer (7, 8). Thus, LKB1 is a tumor suppressor, and the charomyces cerevisiae, the AMPK ortholog is required for the findings that it acted upstream of AMPK introduced the first link response to glucose starvation, especially for the switch from between AMPK and cancer. rapid growth in high glucose using fermentative metabolism The g subunits of AMPK contain three binding sites for AMP, (i.e., glycolysis) to the slower growth using oxidative metabolism with ADP and ATP binding in competition with AMP, at least at that occurs when glucose becomes limiting (3). This metabolic two of them (9, 10). AMP binding activates AMPK by three switch is equivalent to reversal of the Warburg effect that occurs distinct mechanisms: (i) increased Thr172 phosphorylation by in many rapidly proliferating mammalian cells, including tumor LKB1; (ii) decreased Thr172 dephosphorylation by protein phos- cells. phatases; and (iii) caused allosteric activation (>10-fold; ref. 11; ATP and ADP can be likened to the chemicals in a rechargeable Fig. 1). This tripartite mechanism makes the system an exquisitely battery, with a high ratio of ATP:ADP representing a fully charged sensitive sensor of cellular energy status. Effects (ii) and possibly (i), cellular "battery," while any decrease indicates that the battery is but not (iii), are mimicked by binding of ADP, while all three are becoming flat. Because the reaction catalyzed by adenylate kinase antagonized by ATP (11–13). All three of these effects are due to (2ADP $ ATP þ AMP) operates close to equilibrium in most binding of AMP to AMPK itself, rather than to the upstream kinase eukaryotic cells, any increase in ADP:ATP is always accompanied orphosphatase.Thus, althoughLKB1 normallyhas tobepresent for by a much larger rise in AMP:ATP (4), making the latter ratio a cellular energy stress to activate AMPK, it is not itself activated by particularly sensitive indicator of energy stress. AMPK monitors it (14). An alternate upstream kinase phosphorylating Thr172, the calmodulin-dependent kinase CaMKKb (encoded by CAMKK2), þ is only active in cells when intracellular Ca2 has been elevated Division of Cell Signalling and Immunology, College of Life Sciences, (Fig. 1). This alternate, AMP-independent pathway mediates the þ University of Dundee, Dundee, Scotland, United Kingdom. effects of hormones that use Ca2 as a second messenger (15, 16). Corresponding Author: D. Grahame Hardie, Division of Cell Signalling and Once activated by energy stress, AMPK acts to restore energy Immunology, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, homeostasis by promoting catabolic pathways generating ATP, Scotland, United Kingdom. Phone: 44-1382-384253; Fax: 44-1382-385507; while inhibiting ATP-consuming processes (1). The latter include E-mail: [email protected] most anabolic pathways, including those promoted by the doi: 10.1158/1078-0432.CCR-14-3300 mTORC1 signaling pathway, which is inhibited by AMPK Ó2015 American Association for Cancer Research. (17, 18). Because AMPK switches off the synthesis of lipids, RNAs, 3836 Clin Cancer Res; 21(17) September 1, 2015 Downloaded from clincancerres.aacrjournals.org on September 24, 2021. © 2015 American Association for Cancer Research. Published OnlineFirst July 7, 2015; DOI: 10.1158/1078-0432.CCR-14-3300 Is AMPK a Friend or Foe in Cancer? Ca2+ AMP MO25 STRAD ADP? LKB1 Glucose uptake (some cell types) CaMKKb AMP 1 ATP Glycolysis (some cell types) Catabolic Fatty acid oxidation ATP 3 pathways Oxidative metabolism Upstream kinases Mitochondrial biogenesis aaP Autophagy bbgg Upstream Fatty acid synthesis AMPK phosphatases AMPK Triglyceride/phospholipid synthesis Anabolic Sterol synthesis pathways Glycogen synthesis 2 ATP rRNA synthesis Protein synthesis AMP ADP © 2015 American Association for Cancer Research Figure 1. Tripartite mechanism for AMPK activation by 5'-AMP. AMPK is phosphorylated at Thr172 and activated by upstream kinases, especially the constitutively active þ kinase LKB1 (which is only active in complex with MO25 and STRAD) and the Ca2 /calmodulin-dependent kinase kinase, CaMKKb. Binding of AMP to AMPK activates the kinase by three mechanisms, all of which are antagonized by ATP: (1) binding of AMP (and possibly ADP) promotes Thr172 phosphorylation by LKB1; (2) binding of AMP (and ADP at higher concentrations) inhibits Thr172 dephosphorylation by phosphatases; and (3) binding of AMP (but not ADP) causes 10-fold allosteric activation. Once activated by energy stress, AMPK acts to restore energy homeostasis by activating catabolic pathways (including oxidative metabolism) and by inhibiting anabolic pathways (including those downstream of mTORC1). and proteins, it inhibits cell growth. It also causes a G1 cell-cycle As well as being required for activation of AMPK, LKB1 also arrest by promoting phosphorylation of p53, thus blocking DNA activates a family of 12 AMPK-related kinases (ARK) by phosphor- synthesis (19, 20). Although AMPK can acutely enhance glucose ylating the threonine residue equivalent to Thr172 (31). None of uptake and glycolysis in some cell types, in the longer term it these kinases appear to be activated by energy stress or to directly promotes (like its yeast ortholog) the more glucose-sparing, mito- inhibit cell growth and division, and it therefore seems likely that chondrial oxidative metabolism used by quiescent cells, rather most tumor-suppressive effects of LKB1 are mediated by AMPK. than the rapid glucose uptake, glycolysis, and pentose phosphate However, reduced function caused by loss of LKB1 of two of the pathway used predominantly by proliferating cells (21). ARKs, MARK1 and MARK4, does contribute to increased migration Numerous pharmacologic agents that activate AMPK have been and metastasis of epithelial tumor cells in mouse models (32). identified, including many natural plant products, or their deri- vatives, used in traditional medicines (22). These include the – antidiabetic biguanides metformin (23) and phenformin (6), Clinical Translational Advances both derived from the natural product galegine, as well as salic- Loss of a single AMPK-a1 allele accelerates development of ylate, the active component of willow bark, of which acetyl lymphomas induced in mice by transgenic expression of Myc in B salicylic acid (ASA or aspirin) is a synthetic derivative as well as cells, whereas loss of both alleles has an even larger effect (33). a prodrug (24). Metformin, phenformin, and galegine, and many Although this suggests that AMPK can act as a tumor suppressor, natural products such as resveratrol and berberine, activate AMPK mutations in genes encoding AMPK subunits appear to be rather indirectly by inhibiting mitochondrial ATP synthesis, thus infrequent in human cancers. This might either be because of increasing cellular AMP (25). However, salicylate activates AMPK redundancy between AMPK isoforms, or perhaps more likely by direct binding
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