Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells
Abstract Intracellular Ca2+ signals control several physiological and pathophysiological processes. The main tool to chelate intracellular Ca2+ is intracellular BAPTA (BAPTAi), usually introduced into cells as a membrane-permeant acetoxymethyl ester (BAPTA-AM). Previously, we demonstrated that BAPTA...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Publishing Group
2023-09-01
|
| Series: | Cell Death and Disease |
| Online Access: | https://doi.org/10.1038/s41419-023-06120-4 |
| _version_ | 1827821197149077504 |
|---|---|
| author | Flore Sneyers Martijn Kerkhofs Femke Speelman-Rooms Kirsten Welkenhuyzen Rita La Rovere Ahmed Shemy Arnout Voet Guy Eelen Mieke Dewerchin Stephen W. G. Tait Bart Ghesquière Martin D. Bootman Geert Bultynck |
| author_facet | Flore Sneyers Martijn Kerkhofs Femke Speelman-Rooms Kirsten Welkenhuyzen Rita La Rovere Ahmed Shemy Arnout Voet Guy Eelen Mieke Dewerchin Stephen W. G. Tait Bart Ghesquière Martin D. Bootman Geert Bultynck |
| author_sort | Flore Sneyers |
| collection | DOAJ |
| description | Abstract Intracellular Ca2+ signals control several physiological and pathophysiological processes. The main tool to chelate intracellular Ca2+ is intracellular BAPTA (BAPTAi), usually introduced into cells as a membrane-permeant acetoxymethyl ester (BAPTA-AM). Previously, we demonstrated that BAPTAi enhanced apoptosis induced by venetoclax, a BCL-2 antagonist, in diffuse large B-cell lymphoma (DLBCL). This finding implied a novel interplay between intracellular Ca2+ signaling and anti-apoptotic BCL-2 function. Hence, we set out to identify the underlying mechanisms by which BAPTAi enhances cell death in B-cell cancers. In this study, we discovered that BAPTAi alone induced apoptosis in hematological cancer cell lines that were highly sensitive to S63845, an MCL-1 antagonist. BAPTAi provoked a rapid decline in MCL-1-protein levels by inhibiting mTORC1-driven Mcl-1 translation. These events were not a consequence of cell death, as BAX/BAK-deficient cancer cells exhibited similar downregulation of mTORC1 activity and MCL-1-protein levels. Next, we investigated how BAPTAi diminished mTORC1 activity and identified its ability to impair glycolysis by directly inhibiting 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) activity, a previously unknown effect of BAPTAi. Notably, these effects were also induced by a BAPTAi analog with low affinity for Ca2+. Consequently, our findings uncover PFKFB3 inhibition as an Ca2+-independent mechanism through which BAPTAi impairs cellular metabolism and ultimately compromises the survival of MCL-1-dependent cancer cells. These findings hold two important implications. Firstly, the direct inhibition of PFKFB3 emerges as a key regulator of mTORC1 activity and a promising target in MCL-1-dependent cancers. Secondly, cellular effects caused by BAPTAi are not necessarily related to Ca2+ signaling. Our data support the need for a reassessment of the role of Ca2+ in cellular processes when findings were based on the use of BAPTAi. |
| first_indexed | 2024-03-12T01:39:54Z |
| format | Article |
| id | doaj.art-165f5a0201694ead8a06898f90b65886 |
| institution | Directory Open Access Journal |
| issn | 2041-4889 |
| language | English |
| last_indexed | 2024-03-12T01:39:54Z |
| publishDate | 2023-09-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death and Disease |
| spelling | doaj.art-165f5a0201694ead8a06898f90b658862023-09-10T11:28:17ZengNature Publishing GroupCell Death and Disease2041-48892023-09-0114911710.1038/s41419-023-06120-4Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cellsFlore Sneyers0Martijn Kerkhofs1Femke Speelman-Rooms2Kirsten Welkenhuyzen3Rita La Rovere4Ahmed Shemy5Arnout Voet6Guy Eelen7Mieke Dewerchin8Stephen W. G. Tait9Bart Ghesquière10Martin D. Bootman11Geert Bultynck12KU Leuven, Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Campus Gasthuisberg O&N IKU Leuven, Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Campus Gasthuisberg O&N IKU Leuven, Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Campus Gasthuisberg O&N IKU Leuven, Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Campus Gasthuisberg O&N IKU Leuven, Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Campus Gasthuisberg O&N IKU Leuven, Laboratory for Biomolecular Modelling and Design, Department of ChemistryKU Leuven, Laboratory for Biomolecular Modelling and Design, Department of ChemistryKU Leuven, Laboratory of Angiogenesis and Vascular Metabolism, Department of Oncology, Leuven Cancer Institute, Campus Gasthuisberg O&N4KU Leuven, Laboratory of Angiogenesis and Vascular Metabolism, Department of Oncology, Leuven Cancer Institute, Campus Gasthuisberg O&N4Cancer Research UK Beatson Institute, School of Cancer Sciences, University of GlasgowKU Leuven, Laboratory of Applied Mass Spectrometry, Department of Cellular and Molecular Medicine, Leuven, Belgium — VIB, Metabolomics Core Facility Leuven, Center for Cancer Biology, Leuven, BelgiumSchool of Life, Health and Chemical Sciences, Faculty of Science, Technology, Engineering and Mathematics, The Open University, Walton HallKU Leuven, Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Campus Gasthuisberg O&N IAbstract Intracellular Ca2+ signals control several physiological and pathophysiological processes. The main tool to chelate intracellular Ca2+ is intracellular BAPTA (BAPTAi), usually introduced into cells as a membrane-permeant acetoxymethyl ester (BAPTA-AM). Previously, we demonstrated that BAPTAi enhanced apoptosis induced by venetoclax, a BCL-2 antagonist, in diffuse large B-cell lymphoma (DLBCL). This finding implied a novel interplay between intracellular Ca2+ signaling and anti-apoptotic BCL-2 function. Hence, we set out to identify the underlying mechanisms by which BAPTAi enhances cell death in B-cell cancers. In this study, we discovered that BAPTAi alone induced apoptosis in hematological cancer cell lines that were highly sensitive to S63845, an MCL-1 antagonist. BAPTAi provoked a rapid decline in MCL-1-protein levels by inhibiting mTORC1-driven Mcl-1 translation. These events were not a consequence of cell death, as BAX/BAK-deficient cancer cells exhibited similar downregulation of mTORC1 activity and MCL-1-protein levels. Next, we investigated how BAPTAi diminished mTORC1 activity and identified its ability to impair glycolysis by directly inhibiting 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) activity, a previously unknown effect of BAPTAi. Notably, these effects were also induced by a BAPTAi analog with low affinity for Ca2+. Consequently, our findings uncover PFKFB3 inhibition as an Ca2+-independent mechanism through which BAPTAi impairs cellular metabolism and ultimately compromises the survival of MCL-1-dependent cancer cells. These findings hold two important implications. Firstly, the direct inhibition of PFKFB3 emerges as a key regulator of mTORC1 activity and a promising target in MCL-1-dependent cancers. Secondly, cellular effects caused by BAPTAi are not necessarily related to Ca2+ signaling. Our data support the need for a reassessment of the role of Ca2+ in cellular processes when findings were based on the use of BAPTAi.https://doi.org/10.1038/s41419-023-06120-4 |
| spellingShingle | Flore Sneyers Martijn Kerkhofs Femke Speelman-Rooms Kirsten Welkenhuyzen Rita La Rovere Ahmed Shemy Arnout Voet Guy Eelen Mieke Dewerchin Stephen W. G. Tait Bart Ghesquière Martin D. Bootman Geert Bultynck Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells Cell Death and Disease |
| title | Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells |
| title_full | Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells |
| title_fullStr | Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells |
| title_full_unstemmed | Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells |
| title_short | Intracellular BAPTA directly inhibits PFKFB3, thereby impeding mTORC1-driven Mcl-1 translation and killing MCL-1-addicted cancer cells |
| title_sort | intracellular bapta directly inhibits pfkfb3 thereby impeding mtorc1 driven mcl 1 translation and killing mcl 1 addicted cancer cells |
| url | https://doi.org/10.1038/s41419-023-06120-4 |
| work_keys_str_mv | AT floresneyers intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT martijnkerkhofs intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT femkespeelmanrooms intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT kirstenwelkenhuyzen intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT ritalarovere intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT ahmedshemy intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT arnoutvoet intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT guyeelen intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT miekedewerchin intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT stephenwgtait intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT bartghesquiere intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT martindbootman intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells AT geertbultynck intracellularbaptadirectlyinhibitspfkfb3therebyimpedingmtorc1drivenmcl1translationandkillingmcl1addictedcancercells |