Selective Activation of AMPK<i>β</i>1-Containing Isoforms Improves Kidney Function in a Rat Model of Diabetic Nephropathy

Salatto, Christopher T.; Miller, Russell A.; Cameron, Kimberly O.; Cokorinos, Emily; Reyes, Allan R.; Ward, Jessica; Calabrese, Matthew F.; Kurumbail, Ravi G.; Rajamohan, Francis; Kalgutkar, Amit S.; Tess, David A.; Shavnya, Andre; Genung, Nathan E.; Edmonds, David J.; Jatkar, Aditi; Maciejewski, Benjamin S.; Amaro, Marina; Gandhok, Harmeet; Monetti, Mara; Cialdea, Katherine; Bollinger, Eliza; Kreeger, John M.; Coskran, Timothy; Opsahl, Alan; Boucher, Germaine; Birnbaum, Morris J.; DaSilva-Jardine, Paul; Rolph, Tim

doi:10.1124/jpet.116.237925

Cited by 70 publications

(62 citation statements)

References 25 publications

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“…This provided us with cells with defined β-isoform expression, allowing us to investigate β-isoform-specific regulation of AMPK. Cells were treated in the absence or presence of increasing concentrations of two different AMPK activators, 991 or PF06685249 (hereafter referred to as PF249), a β1-selective AMPK activator [31]. In both β1- and β2-expressing cells, 991 caused a dose-dependent increase in Thr 172 and ACC phosphorylation (Figure 3A).…”

Section: Resultsmentioning

confidence: 99%

“…This presumably reflects the fact that, in addition to increasing Thr 172 phosphorylation, 991 causes a significant allosteric activation of AMPK, which would result in more marked phosphorylation of downstream targets. Treatment with PF249, which is more than 1000-fold more potent towards β1- compared with β2-complexes [31], led to a robust increase in ACC phosphorylation, together with a weaker, but detectable, increase in Thr 172 phosphorylation in β1-expressing cells. Importantly, however, PF249 had virtually no effect on Thr 172 or ACC phosphorylation in β2-expressing cells (Figure 3B).…”

Section: Resultsmentioning

confidence: 99%

“…PF06685249 [31] was a generous gift from Dr Russell Miller (Pfizer Global Research and Development). Recombinant PP2Cα was purified as described previously [32].…”

Section: Methodsmentioning

confidence: 99%

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Effect of different γ-subunit isoforms on the regulation of AMPK

Willows

Navaratnam

Lima

et al. 2017

Biochemical Journal

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AMP-activated protein kinase (AMPK) plays a key role in integrating metabolic pathways in response to energy demand. AMPK activation results in a wide range of downstream responses, many of which are associated with improved metabolic outcome, making AMPK an attractive target for the treatment of metabolic diseases. AMPK is a heterotrimeric complex consisting of a catalytic subunit (α) and two regulatory subunits (β and γ). The γ-subunit harbours the nucleotide-binding sites and plays an important role in AMPK regulation in response to cellular energy levels. In mammals, there are three isoforms of the γ-subunit and these respond differently to regulation by nucleotides, but there is limited information regarding their role in activation by small molecules. Here, we determined the effect of different γ-isoforms on AMPK by a direct activator, 991. In cells, 991 led to a greater activation of γ2-containing AMPK complexes compared with either γ1 or γ3. This effect was dependent on the long N-terminal region of the γ2-isoform. We were able to rule out an effect of Ser108 phosphorylation, since mutation of Ser108 to alanine in the β2-isoform had no effect on activation of AMPK by 991 in either γ1- or γ2-complexes. The rate of dephosphorylation of Thr172 was slower for γ2- compared with γ1-complexes, both in the absence and presence of 991. Our studies show that activation of AMPK by 991 depends on the nature of the γ-isoform. This finding may have implications for the design of isoform-selective AMPK activators.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Effect of different γ-subunit isoforms on the regulation of AMPK

Willows

Navaratnam

Lima

et al. 2017

Biochemical Journal

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“…991 activates both β1-and β2containing complexes (with higher affinity to the β1 complexes), thus it acts as a pan/total AMPK activator. Given that isoform-specific AMPK activators have recently been identified 34,58 , it would be of interest to elucidate isoform-specific gene responses and metabolic programming.…”

Section: Discussionmentioning

confidence: 99%

AMPK promotes induction of a tumor suppressor FLCN through activation of TFEB independently of mTOR

Collodet

Foretz

Deák

et al. 2018

Preprint

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AMP-activated protein kinase (AMPK) is a central energy sensor and master regulator of energy homeostasis. AMPK not only elicits acute metabolic responses, but also promotes metabolic reprogramming and adaptions in the long-term through regulation of specific transcription factors/co-activators. We performed a whole-genome transcriptome profiling in wild-type and AMPK-deficient mouse embryonic fibroblasts (MEF) and mouse primary hepatocytes that had been treated with two distinct classes of small-molecule AMPK activators, namely 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) or 991. This led to the identification of distinct compound-dependent gene expression signatures and to the discovery of several AMPK-regulated genes, including folliculin (Flcn), a gene encoding for a tumor suppressor and nutrient sensor. Gene set enrichment and pathway analyses identified the lysosomal pathway and the associated transcription factor EB (TFEB) as key transcriptional mediator responsible for AMPK-dependent gene expression changes. AMPK-induced Flcn expression was abolished in TFEB/TFE3 double knockout MEF and the promoter activity of Flcn was profoundly reduced when its putative TFEB-binding site was mutated.Mechanistically, we have found that AMPK promotes the dephosphorylation and nuclear localization of TFEB independently of mTOR activity.Collectively, we identified the AMPK-TFEB-FLCN axis as a potential key regulator for cellular and metabolic homeostasis. Moreover, data from zebrafish with physiologically and pharmacologically activated AMPK confirmed the AMPK-TFEB-FLCN cascade in vivo.is the tumor suppressor kinase LKB1 11 . In some cell types, Thr172 can be phosphorylated in a Ca 2+ -mediated process catalyzed by Ca 2+ /calmodulin-dependent protein kinase kinases 12 .AMPK is considered an attractive therapeutic target for metabolic disorders since AMPK activation brings about metabolic responses anticipated to counteract the metabolic abnormalities associated with obesity, insulin resistance, and type 2 diabetes 13,14 . Indeed several compounds, which can be divided into three categories 12 , have been reported to activate AMPK and elicit metabolic effects in cellular and pre-clinical studies. The first class comprises indirect activators which, through inhibition of mitochondrial respiration and eventual suppression of ATP synthesis, increase cellular AMP:ATP or ADP:ATP ratios (e.g. metformin, resveratrol) 15 .

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“…While reducing blood glucose therefore requires a pan-β activator, β1-selective activators may be useful for other purposes. For example, PF-06409577 activated AMPK in the kidney, and reduced urinary protein and other markers of kidney damage in a rat model of diabetic kidney disease (13).…”

mentioning

confidence: 99%

Targeting an energy sensor to treat diabetes

Hardie

2017

Science

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The AMP-activated protein kinase (AMPK) is an energy sensor that monitors AMP:ATP and ADP:ATP ratios in living cells (1). Once activated by rises in these ratios (signifying a fall in cellular energy), it acts to restore energy balance by switching on metabolic pathways that generate ATP (catabolism), while switching off processes that consume ATP, including synthesis and storage of macromolecules (anabolism). Not surprisingly, energy balance at the cellular and whole body levels are intimately connected. Obesity, which occurs when whole body energy intake exceeds energy expenditure for prolonged periods, is a major public health issue because it increases the risk of disorders such as Type 2 diabetes. In obese individuals, liver and muscle may store excess fat, leading to resistance to the hormone insulin. Released when blood glucose rises after meals, insulin normally promotes glucose uptake by muscle and represses glucose production by liver, thus rapidly returning

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Selective Activation of AMPKβ1-Containing Isoforms Improves Kidney Function in a Rat Model of Diabetic Nephropathy

Cited by 70 publications

References 25 publications

Effect of different γ-subunit isoforms on the regulation of AMPK

Effect of different γ-subunit isoforms on the regulation of AMPK

AMPK promotes induction of a tumor suppressor FLCN through activation of TFEB independently of mTOR

Targeting an energy sensor to treat diabetes

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