A single nucleotide polymorphism genotyping platform for the authentication of patient derived xenografts
Metrics: PDF 1413 views | HTML 3443 views | ?
Jad El-Hoss1,*, Duohui Jing1,*, Kathryn Evans1, Cara Toscan1, Jinhan Xie1, Hyunjoo Lee1, Renea A. Taylor2, Mitchell G. Lawrence3, Gail P. Risbridger3, Karen L. MacKenzie1, Rosemary Sutton1, Richard B. Lock1
1Children’s Cancer Institute, Lowy Cancer Research Centre, Sydney, UNSW, Australia
2Prostate Research Group, Department of Physiology, Biomedicine Discovery Institute, Monash Partners Comprehensive Cancer Consortium, Monash University, Clayton, VIC, Australia
3Prostate Research Group, Department of Anatomy and Developmental Biology, Biomedicine Discovery Institute, Monash Partners Comprehensive Cancer Consortium, Monash University, Clayton, VIC, Australia
*These authors have contributed equally to this work
Richard B. Lock, email: firstname.lastname@example.org
Keywords: SNP genotyping, patient derived xenografts, authentication, OpenArray, R studio
Received: December 06, 2015 Accepted: July 26, 2016 Published: August 09, 2016
Patient derived xenografts (PDXs) have become a vital, frequently used, component of anti-cancer drug development. PDXs can be serially passaged in vivo for years, and shared across laboratories. As a consequence, the potential for mis-identification and cross-contamination is possible, yet authentication of PDXs appears limited. We present a PDX Authentication System (PAS), by combining a commercially available OpenArray assay of single nucleotide polymorphisms (SNPs) with in-house R studio programs, to validate PDXs established in individual mice from acute lymphoblastic leukemia biopsies. The PAS is sufficiently robust to identify contamination at levels as low as 3%, similar to the gold standard of short tandem repeat (STR) profiling. We have surveyed a panel of PDXs established from 73 individual leukemia patients, and found that the PAS provided sufficient discriminatory power to identify each xenograft. The identified SNP-discrepant PDXs demonstrated distinct gene expression profiles, indicating a risk of contamination for PDXs at high passage number. The PAS also allows for the authentication of tumor cells with complex karyotypes from solid tumors including prostate cancer and Ewing’s sarcoma. This study highlights the demands of authenticating PDXs for cancer research, and evaluates a reliable authentication platform that utilizes a commercially available and cost-effective system.
All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 License.