Highlights
- •The metabolic characteristics of cancer stem cells in cholangiocarcinoma are not known.
- •Cholangiocarcinoma stem-like cells preferentially use oxidative phosphorylation as a source of energy.
- •PGC-1α is a key molecule regulating the metabolic features of cholangiocarcinoma stem-like cells.
- •Interfering with oxidative phosphorylation or PGC-1α limits the development of tumors originating from stem-like cells in vivo.
- •Expression of PGC-1α or proteins of the mitochondrial respiratory complex correlate with clinical outcomes in patients with cholangiocarcinoma.
Background & Aims
Little is known about the metabolic regulation of cancer stem cells (CSCs) in cholangiocarcinoma
(CCA). We analyzed whether mitochondrial-dependent metabolism and related signaling
pathways contribute to stemness in CCA.
Methods
The stem-like subset was enriched by sphere culture (SPH) in human intrahepatic CCA
cells (HUCCT1 and CCLP1) and compared to cells cultured in monolayer. Extracellular
flux analysis was examined by Seahorse technology and high-resolution respirometry.
In patients with CCA, expression of factors related to mitochondrial metabolism was
analyzed for possible correlation with clinical parameters.
Results
Metabolic analyses revealed a more efficient respiratory phenotype in CCA-SPH than
in monolayers, due to mitochondrial oxidative phosphorylation. CCA-SPH showed high
mitochondrial membrane potential and elevated mitochondrial mass, and over-expressed
peroxisome proliferator-activated receptor gamma coactivator (PGC)-1α, a master regulator
of mitochondrial biogenesis. Targeting mitochondrial complex I in CCA-SPH using metformin,
or PGC-1α silencing or pharmacologic inhibition (SR-18292), impaired spherogenicity
and expression of markers related to the CSC phenotype, pluripotency, and epithelial-mesenchymal
transition. In mice with tumor xenografts generated by injection of CCA-SPH, administration
of metformin or SR-18292 significantly reduced tumor growth and determined a phenotype
more similar to tumors originated from cells grown in monolayer. In patients with
CCA, expression of PGC-1α correlated with expression of mitochondrial complex II and
of stem-like genes. Patients with higher PGC-1α expression by immunostaining had lower
overall and progression-free survival, increased angioinvasion and faster recurrence.
In GSEA analysis, patients with CCA and high levels of mitochondrial complex II had
shorter overall survival and time to recurrence.
Conclusions
The CCA stem-subset has a more efficient respiratory phenotype and depends on mitochondrial
oxidative metabolism and PGC-1α to maintain CSC features.
Lay summary
The growth of many cancers is sustained by a specific type of cells with more embryonic
characteristics, termed ‘cancer stem cells’. These cells have been described in cholangiocarcinoma,
a type of liver cancer with poor prognosis and limited therapeutic approaches. We
demonstrate that cancer stem cells in cholangiocarcinoma have different metabolic
features, and use mitochondria, an organelle located within the cells, as the major
source of energy. We also identify PGC-1α, a molecule which regulates the biology
of mitochondria, as a possible new target to be explored for developing new treatments
for cholangiocarcinoma.
Graphical abstract
Graphical Abstract
Keywords
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Article info
Publication history
Published online: January 20, 2021
Accepted:
December 24,
2020
Received in revised form:
December 18,
2020
Received:
March 22,
2020
Footnotes
Author names in bold designate shared co-first authorship
Identification
Copyright
© 2021 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
