Cargando…
Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering
OBJECTIVE: Skeletal muscle is an attractive target for blood glucose-lowering pharmacological interventions. Oral dosing of small molecule direct pan-activators of AMPK that bind to the allosteric drug and metabolite (ADaM) site, lowers blood glucose through effects in skeletal muscle. The molecular...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8381035/ https://www.ncbi.nlm.nih.gov/pubmed/34033941 http://dx.doi.org/10.1016/j.molmet.2021.101259 |
_version_ | 1783741288341831680 |
---|---|
author | Jørgensen, Nicolas O. Kjøbsted, Rasmus Larsen, Magnus R. Birk, Jesper B. Andersen, Nicoline R. Albuquerque, Bina Schjerling, Peter Miller, Russell Carling, David Pehmøller, Christian K. Wojtaszewski, Jørgen F.P. |
author_facet | Jørgensen, Nicolas O. Kjøbsted, Rasmus Larsen, Magnus R. Birk, Jesper B. Andersen, Nicoline R. Albuquerque, Bina Schjerling, Peter Miller, Russell Carling, David Pehmøller, Christian K. Wojtaszewski, Jørgen F.P. |
author_sort | Jørgensen, Nicolas O. |
collection | PubMed |
description | OBJECTIVE: Skeletal muscle is an attractive target for blood glucose-lowering pharmacological interventions. Oral dosing of small molecule direct pan-activators of AMPK that bind to the allosteric drug and metabolite (ADaM) site, lowers blood glucose through effects in skeletal muscle. The molecular mechanisms responsible for this effect are not described in detail. This study aimed to illuminate the mechanisms by which ADaM-site activators of AMPK increase glucose uptake in skeletal muscle. Further, we investigated the consequence of co-stimulating muscles with two types of AMPK activators i.e., ADaM-site binding small molecules and the prodrug AICAR. METHODS: The effect of the ADaM-site binding small molecules (PF739 and 991), AICAR or co-stimulation with PF739 or 991 and AICAR on muscle glucose uptake was investigated ex vivo in m. extensor digitorum longus (EDL) excised from muscle-specific AMPKα1α2 as well as whole-body AMPKγ3-deficient mouse models. In vitro complex-specific AMPK activity was measured by immunoprecipitation and molecular signaling was assessed by western blotting in muscle lysate. To investigate the transferability of these studies, we treated diet-induced obese mice in vivo with PF739 and measured complex-specific AMPK activation in skeletal muscle. RESULTS: Incubation of skeletal muscle with PF739 or 991 increased skeletal muscle glucose uptake in a dose-dependent manner. Co-incubating PF739 or 991 with a maximal dose of AICAR increased glucose uptake to a greater extent than any of the treatments alone. Neither PF739 nor 991 increased AMPKα2β2γ3 activity to the same extent as AICAR, while co-incubation led to potentiated effects on AMPKα2β2γ3 activation. In muscle from AMPKγ3 KO mice, AICAR-stimulated glucose uptake was ablated. In contrast, the effect of PF739 or 991 on glucose uptake was not different between WT and AMPKγ3 KO muscles. In vivo PF739 treatment lowered blood glucose levels and increased muscle AMPKγ1-complex activity 2-fold, while AMPKα2β2γ3 activity was not affected. CONCLUSIONS: ADaM-site binding AMPK activators increase glucose uptake independently of AMPKγ3. Co-incubation with PF739 or 991 and AICAR potentiates the effects on muscle glucose uptake and AMPK activation. In vivo, PF739 lowers blood glucose and selectively activates muscle AMPKγ1-complexes. Collectively, this suggests that pharmacological activation of AMPKγ1-containing complexes in skeletal muscle can increase glucose uptake and can lead to blood glucose lowering. |
format | Online Article Text |
id | pubmed-8381035 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-83810352021-08-30 Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering Jørgensen, Nicolas O. Kjøbsted, Rasmus Larsen, Magnus R. Birk, Jesper B. Andersen, Nicoline R. Albuquerque, Bina Schjerling, Peter Miller, Russell Carling, David Pehmøller, Christian K. Wojtaszewski, Jørgen F.P. Mol Metab Original Article OBJECTIVE: Skeletal muscle is an attractive target for blood glucose-lowering pharmacological interventions. Oral dosing of small molecule direct pan-activators of AMPK that bind to the allosteric drug and metabolite (ADaM) site, lowers blood glucose through effects in skeletal muscle. The molecular mechanisms responsible for this effect are not described in detail. This study aimed to illuminate the mechanisms by which ADaM-site activators of AMPK increase glucose uptake in skeletal muscle. Further, we investigated the consequence of co-stimulating muscles with two types of AMPK activators i.e., ADaM-site binding small molecules and the prodrug AICAR. METHODS: The effect of the ADaM-site binding small molecules (PF739 and 991), AICAR or co-stimulation with PF739 or 991 and AICAR on muscle glucose uptake was investigated ex vivo in m. extensor digitorum longus (EDL) excised from muscle-specific AMPKα1α2 as well as whole-body AMPKγ3-deficient mouse models. In vitro complex-specific AMPK activity was measured by immunoprecipitation and molecular signaling was assessed by western blotting in muscle lysate. To investigate the transferability of these studies, we treated diet-induced obese mice in vivo with PF739 and measured complex-specific AMPK activation in skeletal muscle. RESULTS: Incubation of skeletal muscle with PF739 or 991 increased skeletal muscle glucose uptake in a dose-dependent manner. Co-incubating PF739 or 991 with a maximal dose of AICAR increased glucose uptake to a greater extent than any of the treatments alone. Neither PF739 nor 991 increased AMPKα2β2γ3 activity to the same extent as AICAR, while co-incubation led to potentiated effects on AMPKα2β2γ3 activation. In muscle from AMPKγ3 KO mice, AICAR-stimulated glucose uptake was ablated. In contrast, the effect of PF739 or 991 on glucose uptake was not different between WT and AMPKγ3 KO muscles. In vivo PF739 treatment lowered blood glucose levels and increased muscle AMPKγ1-complex activity 2-fold, while AMPKα2β2γ3 activity was not affected. CONCLUSIONS: ADaM-site binding AMPK activators increase glucose uptake independently of AMPKγ3. Co-incubation with PF739 or 991 and AICAR potentiates the effects on muscle glucose uptake and AMPK activation. In vivo, PF739 lowers blood glucose and selectively activates muscle AMPKγ1-complexes. Collectively, this suggests that pharmacological activation of AMPKγ1-containing complexes in skeletal muscle can increase glucose uptake and can lead to blood glucose lowering. Elsevier 2021-05-23 /pmc/articles/PMC8381035/ /pubmed/34033941 http://dx.doi.org/10.1016/j.molmet.2021.101259 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Jørgensen, Nicolas O. Kjøbsted, Rasmus Larsen, Magnus R. Birk, Jesper B. Andersen, Nicoline R. Albuquerque, Bina Schjerling, Peter Miller, Russell Carling, David Pehmøller, Christian K. Wojtaszewski, Jørgen F.P. Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering |
title | Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering |
title_full | Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering |
title_fullStr | Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering |
title_full_unstemmed | Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering |
title_short | Direct small molecule ADaM-site AMPK activators reveal an AMPKγ3-independent mechanism for blood glucose lowering |
title_sort | direct small molecule adam-site ampk activators reveal an ampkγ3-independent mechanism for blood glucose lowering |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8381035/ https://www.ncbi.nlm.nih.gov/pubmed/34033941 http://dx.doi.org/10.1016/j.molmet.2021.101259 |
work_keys_str_mv | AT jørgensennicolaso directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT kjøbstedrasmus directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT larsenmagnusr directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT birkjesperb directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT andersennicoliner directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT albuquerquebina directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT schjerlingpeter directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT millerrussell directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT carlingdavid directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT pehmøllerchristiank directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering AT wojtaszewskijørgenfp directsmallmoleculeadamsiteampkactivatorsrevealanampkg3independentmechanismforbloodglucoselowering |