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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.
- Source :
-
PloS one [PLoS One] 2016 Jan 05; Vol. 11 (1), pp. e0145100. Date of Electronic Publication: 2016 Jan 05 (Print Publication: 2016). - Publication Year :
- 2016
-
Abstract
- Introduction: 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).<br />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.<br />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).<br />Conclusion: 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.
- Subjects :
- Aged
Animals
Carcinoma, Non-Small-Cell Lung blood supply
Carcinoma, Non-Small-Cell Lung pathology
DNA, Neoplasm genetics
DNA, Neoplasm metabolism
Female
Graft Survival
Humans
Interleukin Receptor Common gamma Subunit deficiency
Interleukin Receptor Common gamma Subunit genetics
Ischemia
Lung Neoplasms blood supply
Lung Neoplasms pathology
Male
Mice, Inbred NOD
Mice, Knockout
Mice, SCID
Middle Aged
Multiplex Polymerase Chain Reaction
RNA Stability
RNA, Neoplasm metabolism
Retrospective Studies
Reverse Transcriptase Polymerase Chain Reaction
Time Factors
Transplantation, Heterologous
Carcinoma, Non-Small-Cell Lung genetics
Lung Neoplasms genetics
RNA, Neoplasm genetics
Tissue Banks
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 11
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 26731692
- Full Text :
- https://doi.org/10.1371/journal.pone.0145100