title: The Influence of Tissue Ischemia Time on RNA Integrity and Patient-Derived Xenografts (PDX) Engraftment Rate in a Non-Small Cell Lung Cancer (NSCLC) Biobank.
abstract: BACKGROUND
  Bio-repositories are invaluable resources to implement translational cancer research and clinical programs. They represent one of the most powerful tools for biomolecular studies of clinically annotated cohorts, but high quality samples are required to generate reliable molecular readouts and functional studies. The objective of our study was to define the impact of cancer tissue ischemia time on RNA and DNA quality, and for the generation of Patient-Derived Xenografts (PDXs).
  
  
  METHODS
  One-hundred thirty-five lung cancer specimens were selected among our Institutional BioBank samples. Associations between different warm (surgical) and cold (ex-vivo) ischemia time ranges and RNA quality or PDXs engraftment rates were assessed. RNA quality was determined by RNA integrity number (RINs) values. Fresh viable tissue fragments were implanted subcutaneously in NSG mice and serially transplanted.
  
  
  RESULTS
  RNAs with a RIN>7 were detected in 51% of the sample (70/135), with values of RIN significantly lower (OR 0.08, P = 0.01) in samples preserved for more than 3 hours before cryopreservation. Higher quality DNA samples had a concomitant high RIN. Sixty-three primary tumors (41 adenocarcinoma) were implanted with an overall engraftment rate of 33%. Both prolonged warm (>2 hours) and ex-vivo ischemia time (>10 hours) were associated to a lower engraftment rate (OR 0.09 P = 0.01 and OR 0.04 P = 0.008, respectively).
  
  
  CONCLUSIONS
  RNA quality and PDXs engraftment rate were adversely affected by prolonged ischemia times. Proper tissue collection and processing reduce failure rate. Overall, NSCLC BioBanking represents an innovative modality, which can be successfully executed in routine clinical settings, when stringent Standard Operating Procedures are adopted.
authors.affiliation: Department of Thoracic Surgery, University of Torino, 10126, Torino, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Guerrera
authors.affiliation: Department of Molecular Biotechnology and Health Science and Center for Experimental Research and Medical Studies, University of Torino, 10126, Torino, Italy.|Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, 10021, United States of America.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Tabbò
authors.affiliation: Department of Molecular Biotechnology and Health Science and Center for Experimental Research and Medical Studies, University of Torino, 10126, Torino, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Bessone
authors.affiliation: Department of Medical Sciences, University of Torino, 10126, Torino, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Maletta
authors.affiliation: Department of Molecular Biotechnology and Health Science and Center for Experimental Research and Medical Studies, University of Torino, 10126, Torino, Italy.|Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, 10021, United States of America.
authors.forename: Marcello
authors.initials: M
authors.lastname: Gaudiano
authors.affiliation: Department of Molecular Biotechnology and Health Science and Center for Experimental Research and Medical Studies, University of Torino, 10126, Torino, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Ercole
authors.affiliation: Department of Medical Sciences, University of Torino, 10126, Torino, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Annaratone
authors.affiliation: Department of Molecular Biotechnology and Health Science and Center for Experimental Research and Medical Studies, University of Torino, 10126, Torino, Italy.|Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, 10021, United States of America.
authors.forename: Maria
authors.initials: M
authors.lastname: Todaro
authors.affiliation: Department of Medical Sciences, Allergology and Immunology, University of Torino, 10126, Torino, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Boita
authors.affiliation: Department of Thoracic Surgery, University of Torino, 10126, Torino, Italy.
authors.forename: Pier Luigi
authors.initials: PL
authors.lastname: Filosso
authors.affiliation: Unit of Pulmonology, San Giovanni Battista Hospital, 10126, Torino, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Solidoro
authors.affiliation: Department of Medical Sciences, University of Torino, 10126, Torino, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Delsedime
authors.affiliation: Department of Thoracic Surgery, University of Torino, 10126, Torino, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Oliaro
authors.affiliation: Department of Medical Sciences, University of Torino, 10126, Torino, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Sapino
authors.affiliation: Department of Thoracic Surgery, University of Torino, 10126, Torino, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Ruffini
authors.affiliation: Department of Molecular Biotechnology and Health Science and Center for Experimental Research and Medical Studies, University of Torino, 10126, Torino, Italy.|Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, 10021, United States of America.|Department of Pathology and NYU Cancer Center, New York University School of Medicine, New York, NY, 10016, United States of America.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Inghirami
mesh_terms: D000368:Aged; D000818:Animals; D002289:Carcinoma, Non-Small-Cell Lung / Q000098:blood supply / Q000235:genetics* / Q000473:pathology; D004273:DNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D005260:Female; D006085:Graft Survival; D006801:Humans; D053631:Interleukin Receptor Common gamma Subunit / Q000172:deficiency / Q000235:genetics; D007511:Ischemia; D008175:Lung Neoplasms / Q000098:blood supply / Q000235:genetics* / Q000473:pathology; D008297:Male; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction; D020871:RNA Stability; D012334:RNA, Neoplasm / Q000235:genetics* / Q000378:metabolism; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014015:Tissue Banks; D014183:Transplantation, Heterologous
chemical_list: D004273:DNA, Neoplasm; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit; D012334:RNA, Neoplasm