Hannah R Shrader1, Ann M Miller1, Ann Tomanek-Chalkley1, Ashley McCarthy2, Kristen L Coleman2, Po Hien Ear3, Ashutosh K Mangalam4, Aliasger K Salem5, Carlos H F Chan6. 1. Department of Surgery, University of Iowa, Iowa City, IA. 2. Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA. 3. Department of Surgery, University of Iowa, Iowa City, IA; Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA. 4. Department of Pathology, University of Iowa, Iowa City, IA; Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA. 5. Department of Pharmaceutical Sciences and Experimental Therapeutics, University of Iowa, Iowa City, IA; Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA. 6. Department of Surgery, University of Iowa, Iowa City, IA; Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA. Electronic address: carloshfchan@gmail.com.
Abstract
BACKGROUND: Introduction of gut flora into the biliary system is common owing to biliary stenting in patients with obstructing pancreatic head cancer. We hypothesize that alteration of biliary microbiome modifies bile content that modulates pancreatic cancer cell survival. METHODS: Human bile samples were collected during pancreaticoduodenectomy. Bacterial strains were isolated from contaminated (stented) bile and identified using 16S ribosomal RNA sequencing. Human pancreatic cancer cells (AsPC1, CFPAC, Panc1) were treated for 24 hours with sterile (nonstented) bile, contaminated (stented) bile, and sterile bile preincubated with 106 colony forming unit of live bacteria isolated from contaminated bile or a panel of bile acids for 24 hours at 37°C, and evaluated using CellTiter-Blue Cell Viability Assay (Promega Corp. Madison, WI). Human bile (30-50 μl/mouse) was coinjected intraperitoneally with 105 Panc02 mouse pancreatic cancer cells in C57BL6/N mice to evaluate the impact of bile on peritoneal metastasis 3 to 4 weeks after tumor challenge. RESULTS: While all bile samples significantly reduced peritoneal metastasis of Panc02 cells in mice, some contaminated bile samples had diminished antitumor effect. All sterile bile (n = 4) reduced pancreatic cancer cell survival in vitro. Only 40% (2/5) of contaminated bile samples had significant effect. Preincubation of sterile bile with live Enterococcus faecalis or Streptococcus oralis modified the antitumor effect of sterile bile. These changes were not observed with culture media preincubated with live bacteria, suggesting live gut bacteria can modify the antitumor components present in bile. Conjugated bile acids were more potent than unconjugated cholic acid in reducing pancreatic cancer cell survival. CONCLUSION: Alteration of bile microbiome from biliary stenting has a direct impact on pancreatic cancer cell survival. Further study is warranted to determine if this microbiome shift alters tumor microenvironment.
BACKGROUND: Introduction of gut flora into the biliary system is common owing to biliary stenting in patients with obstructing pancreatic head cancer. We hypothesize that alteration of biliary microbiome modifies bile content that modulates pancreatic cancer cell survival. METHODS: Human bile samples were collected during pancreaticoduodenectomy. Bacterial strains were isolated from contaminated (stented) bile and identified using 16S ribosomal RNA sequencing. Human pancreatic cancer cells (AsPC1, CFPAC, Panc1) were treated for 24 hours with sterile (nonstented) bile, contaminated (stented) bile, and sterile bile preincubated with 106 colony forming unit of live bacteria isolated from contaminated bile or a panel of bile acids for 24 hours at 37°C, and evaluated using CellTiter-Blue Cell Viability Assay (Promega Corp. Madison, WI). Human bile (30-50 μl/mouse) was coinjected intraperitoneally with 105 Panc02 mouse pancreatic cancer cells in C57BL6/N mice to evaluate the impact of bile on peritoneal metastasis 3 to 4 weeks after tumor challenge. RESULTS: While all bile samples significantly reduced peritoneal metastasis of Panc02 cells in mice, some contaminated bile samples had diminished antitumor effect. All sterile bile (n = 4) reduced pancreatic cancer cell survival in vitro. Only 40% (2/5) of contaminated bile samples had significant effect. Preincubation of sterile bile with live Enterococcus faecalis or Streptococcus oralis modified the antitumor effect of sterile bile. These changes were not observed with culture media preincubated with live bacteria, suggesting live gut bacteria can modify the antitumor components present in bile. Conjugated bile acids were more potent than unconjugated cholic acid in reducing pancreatic cancer cell survival. CONCLUSION: Alteration of bile microbiome from biliary stenting has a direct impact on pancreatic cancer cell survival. Further study is warranted to determine if this microbiome shift alters tumor microenvironment.
Authors: Claire Jenkins; Clare L Ling; Holly L Ciesielczuk; Julianne Lockwood; Susan Hopkins; Timothy D McHugh; Stephen H Gillespie; Christopher C Kibbler Journal: J Med Microbiol Date: 2011-12-08 Impact factor: 2.472
Authors: Y Lu; M Onda; E Uchida; S Yamamura; K Yanagi; A Matsushita; T Kobayashi; M Fukuhara; K Aida; T Tajiri Journal: Surg Today Date: 2000 Impact factor: 2.549
Authors: Erick Riquelme; Yu Zhang; Liangliang Zhang; Maria Montiel; Michelle Zoltan; Wenli Dong; Pompeyo Quesada; Ismet Sahin; Vidhi Chandra; Anthony San Lucas; Paul Scheet; Hanwen Xu; Samir M Hanash; Lei Feng; Jared K Burks; Kim-Anh Do; Christine B Peterson; Deborah Nejman; Ching-Wei D Tzeng; Michael P Kim; Cynthia L Sears; Nadim Ajami; Joseph Petrosino; Laura D Wood; Anirban Maitra; Ravid Straussman; Matthew Katz; James Robert White; Robert Jenq; Jennifer Wargo; Florencia McAllister Journal: Cell Date: 2019-08-08 Impact factor: 41.582
Authors: Smruti Pushalkar; Mautin Hundeyin; Donnele Daley; Constantinos P Zambirinis; Emma Kurz; Ankita Mishra; Navyatha Mohan; Berk Aykut; Mykhaylo Usyk; Luisana E Torres; Gregor Werba; Kevin Zhang; Yuqi Guo; Qianhao Li; Neha Akkad; Sarah Lall; Benjamin Wadowski; Johana Gutierrez; Juan Andres Kochen Rossi; Jeremy W Herzog; Brian Diskin; Alejandro Torres-Hernandez; Josh Leinwand; Wei Wang; Pardeep S Taunk; Shivraj Savadkar; Malvin Janal; Anjana Saxena; Xin Li; Deirdre Cohen; R Balfour Sartor; Deepak Saxena; George Miller Journal: Cancer Discov Date: 2018-03-22 Impact factor: 39.397
Authors: Amy I Yu; Lili Zhao; Kathryn A Eaton; Sharon Ho; Jiachen Chen; Sara Poe; James Becker; Allison Gonzalez; Delaney McKinstry; Muneer Hasso; Jonny Mendoza-Castrejon; Joel Whitfield; Charles Koumpouras; Patrick D Schloss; Eric C Martens; Grace Y Chen Journal: Cell Rep Date: 2020-04-07 Impact factor: 9.423
Authors: Nabeel Merali; Tarak Chouari; Kayani Kayani; Charles J Rayner; José I Jiménez; Jonathan Krell; Elisa Giovannetti; Izhar Bagwan; Kate Relph; Timothy A Rockall; Tony Dhillon; Hardev Pandha; Nicola E Annels; Adam E Frampton Journal: Cancers (Basel) Date: 2022-02-17 Impact factor: 6.639