TGF-β-induced stromal CYR61 promotes resistance to gemcitabine in pancreatic ductal adenocarcinoma through downregulation of the nucleoside transporters hENT1 and hCNT3.
Pancreatic ductal adenocarcinoma (PDAC) is a lethal cancer in part due to inherent resistance to chemotherapy, including the first-line drug gemcitabine. Although low expression of the nucleoside transporters hENT1 and hCNT3 that mediate cellular uptake of gemcitabine has been linked to gemcitabine resistance, the mechanisms regulating their expression in the PDAC tumor microenvironment are largely unknown. Here, we report that the matricellular protein cysteine-rich angiogenic inducer 61 (CYR61) negatively regulates the nucleoside transporters hENT1 and hCNT3. CRISPR/Cas9-mediated knockout of CYR61 increased expression of hENT1 and hCNT3, increased cellular uptake of gemcitabine and sensitized PDAC cells to gemcitabine-induced apoptosis. In PDAC patient samples, expression of hENT1 and hCNT3 negatively correlates with expression of CYR61 . We demonstrate that stromal pancreatic stellate cells (PSCs) are a source of CYR61 within the PDAC tumor microenvironment. Transforming growth factor-β (TGF-β) induces the expression of CYR61 in PSCs through canonical TGF-β-ALK5-Smad2/3 signaling. Activation of TGF-β signaling or expression of CYR61 in PSCs promotes resistance to gemcitabine in PDAC cells in an in vitro co-culture assay. Our results identify CYR61 as a TGF-β-induced stromal-derived factor that regulates gemcitabine sensitivity in PDAC and suggest that targeting CYR61 may improve chemotherapy response in PDAC patients.
Department of Pharmacology and Cancer Biology.
Rachel A
RA
Hesler
Department of Pharmacology and Cancer Biology.
Jennifer J
JJ
Huang
Division of Medical Oncology, Department of Medicine.
Mark D
MD
Starr
Division of Medical Oncology, Department of Medicine.
Victoria M
VM
Treboschi
Division of Medical Oncology, Department of Medicine.
Alyssa G
AG
Bernanke
Division of Medical Oncology, Department of Medicine.
Andrew B
AB
Nixon
Department of Pathology and.
Shannon J
SJ
McCall
Department of Surgery, Duke University, B354 LSRC Research Drive , Box 91004, Durham, NC 27708 , USA.
Rebekah R
RR
White
Department of Pharmacology and Cancer Biology.|Division of Medical Oncology, Department of Medicine.
Gerard C
GC
Blobe
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy / Q000378:metabolism* / Q000401:mortality; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D055515:Cysteine-Rich Protein 61 / Q000235:genetics* / Q000378:metabolism; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm; D004354:Drug Screening Assays, Antitumor; D033721:Equilibrative Nucleoside Transporter 1 / Q000378:metabolism*; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D026901:Membrane Transport Proteins / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000378:metabolism* / Q000401:mortality; D058954:Pancreatic Stellate Cells / Q000378:metabolism; D015398:Signal Transduction; D018709:Statistics, Nonparametric; D016019:Survival Analysis; D016212:Transforming Growth Factor beta / Q000502:physiology; D059016:Tumor Microenvironment; D000093542:Gemcitabine
D000964:Antimetabolites, Antineoplastic; C501848:CCN1 protein, human; D055515:Cysteine-Rich Protein 61; D033721:Equilibrative Nucleoside Transporter 1; D026901:Membrane Transport Proteins; C438649:SLC29A1 protein, human; D016212:Transforming Growth Factor beta; C065171:cif nucleoside transporter; D003841:Deoxycytidine; D000093542:Gemcitabine