CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo

Abstract Epithelial ovarian cancer (EOC) metastasizes predominantly through multicellular aggregates known as spheroids, which disseminate within the peritoneal cavity and initiate secondary disease upon reattachment at distant sites. EOC spheroids resist detachment-induced cell death by upregulatin...

Full description

Saved in:
Bibliographic Details
Main Authors: Matthew J. Borrelli, Adrian Buensuceso, Yudith Ramos Valdes, Tiffany P.A. Johnston, Jacob Haagsma, Trevor G. Shepherd
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
Subjects:
Online Access:https://doi.org/10.1038/s41598-025-11584-9
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849763509629353984
author Matthew J. Borrelli
Adrian Buensuceso
Yudith Ramos Valdes
Tiffany P.A. Johnston
Jacob Haagsma
Trevor G. Shepherd
author_facet Matthew J. Borrelli
Adrian Buensuceso
Yudith Ramos Valdes
Tiffany P.A. Johnston
Jacob Haagsma
Trevor G. Shepherd
author_sort Matthew J. Borrelli
collection DOAJ
description Abstract Epithelial ovarian cancer (EOC) metastasizes predominantly through multicellular aggregates known as spheroids, which disseminate within the peritoneal cavity and initiate secondary disease upon reattachment at distant sites. EOC spheroids resist detachment-induced cell death by upregulating stress responses including AMP-activated protein kinase (AMPK) signaling and AMPK-dependent macroautophagy (autophagy), highlighting these pathways as potential therapeutic targets. Previously, we used a pharmacological approach to putatively identify Ca2+/calmodulin-dependent protein kinase kinase 2 (CAMKKβ, encoded by CAMKK2) as the primary activator of AMPK in EOC spheroids. Herein we have generated CAMKK2 knockout EOC cell lines via CRISPR–Cas9 genome editing to confirm this function of CAMKKβ and explore the impacts of its loss using in vitro and in vivo models of metastatic EOC. CAMKK2 knockout spheroids exhibited decreased AMPK activation, autophagic flux, cell viability, and metastatic potential relative to parental spheroids, and intraperitoneal xenograft tumours lacking CAMKKβ grew slower than their CAMKKβ-intact counterparts. Effect magnitudes varied between cell line models, suggesting context-dependent roles for CAMKKβ in EOC and rationalizing further studies to characterize the underlying mechanisms. Altogether, our findings highlight CAMKKβ as an important contributor to metabolic reprogramming in EOC spheroids and as a potential therapeutic target in the setting of advanced disease.
format Article
id doaj-art-be8c2a553974454fbfa1be8b7c176fdb
institution DOAJ
issn 2045-2322
language English
publishDate 2025-07-01
publisher Nature Portfolio
record_format Article
series Scientific Reports
spelling doaj-art-be8c2a553974454fbfa1be8b7c176fdb2025-08-20T03:05:23ZengNature PortfolioScientific Reports2045-23222025-07-0115111310.1038/s41598-025-11584-9CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivoMatthew J. Borrelli0Adrian Buensuceso1Yudith Ramos Valdes2Tiffany P.A. Johnston3Jacob Haagsma4Trevor G. Shepherd5The Mary and John Knight Translational Ovarian Cancer Research Unit, Verspeeten Family Cancer CentreThe Mary and John Knight Translational Ovarian Cancer Research Unit, Verspeeten Family Cancer CentreThe Mary and John Knight Translational Ovarian Cancer Research Unit, Verspeeten Family Cancer CentreThe Mary and John Knight Translational Ovarian Cancer Research Unit, Verspeeten Family Cancer CentreThe Mary and John Knight Translational Ovarian Cancer Research Unit, Verspeeten Family Cancer CentreThe Mary and John Knight Translational Ovarian Cancer Research Unit, Verspeeten Family Cancer CentreAbstract Epithelial ovarian cancer (EOC) metastasizes predominantly through multicellular aggregates known as spheroids, which disseminate within the peritoneal cavity and initiate secondary disease upon reattachment at distant sites. EOC spheroids resist detachment-induced cell death by upregulating stress responses including AMP-activated protein kinase (AMPK) signaling and AMPK-dependent macroautophagy (autophagy), highlighting these pathways as potential therapeutic targets. Previously, we used a pharmacological approach to putatively identify Ca2+/calmodulin-dependent protein kinase kinase 2 (CAMKKβ, encoded by CAMKK2) as the primary activator of AMPK in EOC spheroids. Herein we have generated CAMKK2 knockout EOC cell lines via CRISPR–Cas9 genome editing to confirm this function of CAMKKβ and explore the impacts of its loss using in vitro and in vivo models of metastatic EOC. CAMKK2 knockout spheroids exhibited decreased AMPK activation, autophagic flux, cell viability, and metastatic potential relative to parental spheroids, and intraperitoneal xenograft tumours lacking CAMKKβ grew slower than their CAMKKβ-intact counterparts. Effect magnitudes varied between cell line models, suggesting context-dependent roles for CAMKKβ in EOC and rationalizing further studies to characterize the underlying mechanisms. Altogether, our findings highlight CAMKKβ as an important contributor to metabolic reprogramming in EOC spheroids and as a potential therapeutic target in the setting of advanced disease.https://doi.org/10.1038/s41598-025-11584-9Epithelial ovarian cancerSpheroidMetastasisCAMKK2AMPKAutophagy
spellingShingle Matthew J. Borrelli
Adrian Buensuceso
Yudith Ramos Valdes
Tiffany P.A. Johnston
Jacob Haagsma
Trevor G. Shepherd
CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo
Scientific Reports
Epithelial ovarian cancer
Spheroid
Metastasis
CAMKK2
AMPK
Autophagy
title CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo
title_full CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo
title_fullStr CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo
title_full_unstemmed CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo
title_short CAMKKβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo
title_sort camkkβ supports growth and viability of epithelial ovarian cancer in vitro and in vivo
topic Epithelial ovarian cancer
Spheroid
Metastasis
CAMKK2
AMPK
Autophagy
url https://doi.org/10.1038/s41598-025-11584-9
work_keys_str_mv AT matthewjborrelli camkkbsupportsgrowthandviabilityofepithelialovariancancerinvitroandinvivo
AT adrianbuensuceso camkkbsupportsgrowthandviabilityofepithelialovariancancerinvitroandinvivo
AT yudithramosvaldes camkkbsupportsgrowthandviabilityofepithelialovariancancerinvitroandinvivo
AT tiffanypajohnston camkkbsupportsgrowthandviabilityofepithelialovariancancerinvitroandinvivo
AT jacobhaagsma camkkbsupportsgrowthandviabilityofepithelialovariancancerinvitroandinvivo
AT trevorgshepherd camkkbsupportsgrowthandviabilityofepithelialovariancancerinvitroandinvivo