Modulation of Biliary Cancer Chemo-Resistance Through MicroRNA-Mediated Rewiring of the Expansion of CD133+ Cells.
Antineoplastic Agents
/ pharmacology
Biliary Tract Neoplasms
/ drug therapy
CRISPR-Cas Systems
Cell Line, Tumor
Cell Proliferation
/ drug effects
Cell Survival
/ drug effects
Cholangiocarcinoma
/ drug therapy
Cisplatin
/ pharmacology
Deoxycytidine
/ analogs & derivatives
Drug Discovery
Drug Resistance, Neoplasm
/ genetics
Gene Expression Regulation, Neoplastic
High-Throughput Screening Assays
/ methods
Humans
MicroRNAs
/ antagonists & inhibitors
Xenograft Model Antitumor Assays
Gemcitabine
Journal
Hepatology (Baltimore, Md.)
ISSN: 1527-3350
Titre abrégé: Hepatology
Pays: United States
ID NLM: 8302946
Informations de publication
Date de publication:
09 2020
09 2020
Historique:
received:
31
07
2019
accepted:
15
12
2019
pubmed:
28
12
2019
medline:
30
4
2021
entrez:
28
12
2019
Statut:
ppublish
Résumé
Changes in single microRNA (miRNA) expression have been associated with chemo-resistance in biliary tract cancers (BTCs). However, a global assessment of the dynamic role of the microRNome has never been performed to identify potential therapeutic targets that are functionally relevant in the BTC cell response to chemotherapy. High-throughput screening (HTS) of 997 locked nucleic acid miRNA inhibitors was performed in six cholangiocarcinoma cell lines treated with cisplatin and gemcitabine (CG) seeking changes in cell viability. Validation experiments were performed with mirVana probes. MicroRNA and gene expression was assessed by TaqMan assay, RNA-sequencing, and in situ hybridization in four independent cohorts of human BTCs. Knockout of microRNA was achieved by CRISPR-CAS9 in CCLP cells (MIR1249KO) and tested for effects on chemotherapy sensitivity in vitro and in vivo. HTS revealed that MIR1249 inhibition enhanced chemotherapy sensitivity across all cell lines. MIR1249 expression was increased in 41% of cases in human BTCs. In validation experiments, MIR1249 inhibition did not alter cell viability in untreated or dimethyl sulfoxide-treated cells; however, it did increase the CG effect. MIR1249 expression was increased in CD133+ biliary cancer cells freshly isolated from the stem cell niche of human BTCs as well as in CD133+ chemo-resistant CCLP cells. MIR1249 modulated the chemotherapy-induced enrichment of CD133+ cells by controlling their clonal expansion through the Wnt-regulator FZD8. MIR1249KO cells had impaired expansion of the CD133+ subclone and its enrichment after chemotherapy, reduced expression of cancer stem cell markers, and increased chemosensitivity. MIR1249KO xenograft BTC models showed tumor shrinkage after exposure to weekly CG, whereas wild-type models showed only stable disease over treatment. MIR1249 mediates resistance to CG in BTCs and may be tested as a target for therapeutics.
Sections du résumé
BACKGROUND AND AIMS
Changes in single microRNA (miRNA) expression have been associated with chemo-resistance in biliary tract cancers (BTCs). However, a global assessment of the dynamic role of the microRNome has never been performed to identify potential therapeutic targets that are functionally relevant in the BTC cell response to chemotherapy.
APPROACH AND RESULTS
High-throughput screening (HTS) of 997 locked nucleic acid miRNA inhibitors was performed in six cholangiocarcinoma cell lines treated with cisplatin and gemcitabine (CG) seeking changes in cell viability. Validation experiments were performed with mirVana probes. MicroRNA and gene expression was assessed by TaqMan assay, RNA-sequencing, and in situ hybridization in four independent cohorts of human BTCs. Knockout of microRNA was achieved by CRISPR-CAS9 in CCLP cells (MIR1249KO) and tested for effects on chemotherapy sensitivity in vitro and in vivo. HTS revealed that MIR1249 inhibition enhanced chemotherapy sensitivity across all cell lines. MIR1249 expression was increased in 41% of cases in human BTCs. In validation experiments, MIR1249 inhibition did not alter cell viability in untreated or dimethyl sulfoxide-treated cells; however, it did increase the CG effect. MIR1249 expression was increased in CD133+ biliary cancer cells freshly isolated from the stem cell niche of human BTCs as well as in CD133+ chemo-resistant CCLP cells. MIR1249 modulated the chemotherapy-induced enrichment of CD133+ cells by controlling their clonal expansion through the Wnt-regulator FZD8. MIR1249KO cells had impaired expansion of the CD133+ subclone and its enrichment after chemotherapy, reduced expression of cancer stem cell markers, and increased chemosensitivity. MIR1249KO xenograft BTC models showed tumor shrinkage after exposure to weekly CG, whereas wild-type models showed only stable disease over treatment.
CONCLUSIONS
MIR1249 mediates resistance to CG in BTCs and may be tested as a target for therapeutics.
Identifiants
pubmed: 31879968
doi: 10.1002/hep.31094
pmc: PMC7590111
doi:
Substances chimiques
Antineoplastic Agents
0
MIRN1249 microRNA, human
0
MicroRNAs
0
Deoxycytidine
0W860991D6
Cisplatin
Q20Q21Q62J
Gemcitabine
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
982-996Subventions
Organisme : Department of Health
ID : BRC to ICR/Marsden
Pays : United Kingdom
Organisme : Cancer Research UK
ID : CDF TO NV
Pays : United Kingdom
Organisme : Cancer Research UK
ID : CRUK FUNDED CANCER THERAPEUTICS UNIT AT ICR
Pays : United Kingdom
Organisme : Cancer Research UK
ID : PW is a CRUK Life Fellow
Pays : United Kingdom
Informations de copyright
© 2020 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.
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