title: Fibroblast drug scavenging increases intratumoural gemcitabine accumulation in murine pancreas cancer.
abstract: Desmoplasia and hypovascularity are thought to impede drug delivery in pancreatic ductal adenocarcinoma (PDAC). However, stromal depletion approaches have failed to show clinical responses in patients. Here, we aimed to revisit the role of the tumour microenvironment as a physical barrier for gemcitabine delivery.
  
  
  
  Gemcitabine metabolites were analysed inLSL-KrasG12D/+;LSL-Trp53R172H/+;Pdx-1-Cre(KPC) murine tumours and matched liver metastases, primary tumour cell lines, cancer-associated fibroblasts (CAFs) and pancreatic stellate cells (PSCs) by liquid chromatography-mass spectrometry/mass spectrometry. Functional and preclinical experiments, as well as expression analysis of stromal markers and gemcitabine metabolism pathways were performed in murine and human specimen to investigate the preclinical implications and the mechanism of gemcitabine accumulation.
  
  
  
  Gemcitabine accumulation was significantly enhanced in fibroblast-rich tumours compared with liver metastases and normal liver. In vitro, significantly increased concentrations of activated 2',2'-difluorodeoxycytidine-5'-triphosphate (dFdCTP) and greatly reduced amounts of the inactive gemcitabine metabolite 2',2'-difluorodeoxyuridine were detected in PSCs and CAFs. Mechanistically, key metabolic enzymes involved in gemcitabine inactivation such as hydrolytic cytosolic 5'-nucleotidases (Nt5c1A, Nt5c3) were expressed at low levels in CAFs in vitro and in vivo, and recombinant expression of Nt5c1A resulted in decreased intracellular dFdCTP concentrations in vitro. Moreover, gemcitabine treatment in KPC mice reduced the number of liver metastases by >50%.
  
  
  
  Our findings suggest that fibroblast drug scavenging may contribute to the clinical failure of gemcitabine in desmoplastic PDAC. Metabolic targeting of CAFs may thus be a promising strategy to enhance the antiproliferative effects of gemcitabine.
authors.affiliation: Department Gastroenterology and Gastrointestinal Oncology, University Medical Centre Goettingen, Goettingen, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Hessmann
authors.affiliation: Department Gastroenterology and Gastrointestinal Oncology, University Medical Centre Goettingen, Goettingen, Germany.
authors.forename: M S
authors.initials: MS
authors.lastname: Patzak
authors.affiliation: Department Gastroenterology and Gastrointestinal Oncology, University Medical Centre Goettingen, Goettingen, Germany.
authors.forename: L
authors.initials: L
authors.lastname: Klein
authors.affiliation: Department Gastroenterology and Gastrointestinal Oncology, University Medical Centre Goettingen, Goettingen, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Chen
authors.affiliation: Department of General, Visceral and Pediatric Surgery, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Kari
authors.affiliation: Department Gastroenterology and Gastrointestinal Oncology, University Medical Centre Goettingen, Goettingen, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Ramu
authors.affiliation: Cancer Research UK Cambridge Institute, The University of Cambridge, Li Ka Shing Centre, Cambridge, UK.|Oncology iMED DMPK AstraZeneca UK Ltd, HODGKIN C/o B310 Cambridge Science Park, Cambridge, UK.
authors.forename: T E
authors.initials: TE
authors.lastname: Bapiro
authors.affiliation: The University of Manchester, Cancer Research UK Manchester Institute, Manchester, UK.
authors.forename: K K
authors.initials: KK
authors.lastname: Frese
authors.affiliation: Cancer Research UK Cambridge Institute, The University of Cambridge, Li Ka Shing Centre, Cambridge, UK.
authors.forename: A
authors.initials: A
authors.lastname: Gopinathan
authors.affiliation: Cancer Research UK Cambridge Institute, The University of Cambridge, Li Ka Shing Centre, Cambridge, UK.
authors.forename: F M
authors.initials: FM
authors.lastname: Richards
authors.affiliation: Cancer Research UK Cambridge Institute, The University of Cambridge, Li Ka Shing Centre, Cambridge, UK.|Department of Oncology, University of Cambridge, Cambridge, UK.
authors.forename: D I
authors.initials: DI
authors.lastname: Jodrell
authors.affiliation: Department of Pathology, Karolinska University Hospital, Stockholm, Sweden.|Department of Pathology, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
authors.forename: C
authors.initials: C
authors.lastname: Verbeke
authors.affiliation: Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet and Center for Digestive Diseases, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet and Center for Digestive Diseases, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: R
authors.initials: R
authors.lastname: Heuchel
authors.affiliation: Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet and Center for Digestive Diseases, Karolinska University Hospital, Stockholm, Sweden.
authors.forename: J M
authors.initials: JM
authors.lastname: Löhr
authors.affiliation: Department of General, Visceral and Pediatric Surgery, University Medical Center Goettingen, Goettingen, Germany.
authors.forename: S A
authors.initials: SA
authors.lastname: Johnsen
authors.affiliation: Department of Gastroenterology, Endocrinology and Metabolism, Philipps University Marburg, Marburg, Germany.
authors.forename: T M
authors.initials: TM
authors.lastname: Gress
authors.affiliation: Department Gastroenterology and Gastrointestinal Oncology, University Medical Centre Goettingen, Goettingen, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Ellenrieder
authors.affiliation: Department Gastroenterology and Gastrointestinal Oncology, University Medical Centre Goettingen, Goettingen, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Neesse
mesh_terms: D015720:5'-Nucleotidase / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000493:pharmacokinetics* / Q000627:therapeutic use; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy / Q000378:metabolism* / Q000556:secondary; D045744:Cell Line, Tumor; D003570:Cytidine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000627:therapeutic use; D005347:Fibroblasts / Q000378:metabolism*; D005467:Floxuridine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000556:secondary; D051379:Mice; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D061251:Primary Cell Culture; D059016:Tumor Microenvironment; D000093542:Gemcitabine
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D000964:Antimetabolites, Antineoplastic; D005467:Floxuridine; D003841:Deoxycytidine; C054403:2',2'-difluorodeoxycytidine 5'-triphosphate; D003570:Cytidine Triphosphate; D015720:5'-Nucleotidase; C000621979:NT5C1A protein, mouse; C503010:Nt5c3 protein, mouse; C084565:2',2'-difluoro-2'-deoxyuridine; D000093542:Gemcitabine
keywords: CHEMOTHERAPY; DRUG METABOLISM; PANCREATIC CANCER; PANCREATIC FIBROSIS