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Deep Clonal Profiling of Formalin Fixed Paraffin Embedded Clinical Samples
Authors:Tara Holley  Elizabeth Lenkiewicz  Lisa Evers  Waibhav Tembe  Christian Ruiz  Joel R Gsponer  Cyrill A Rentsch  Lukas Bubendorf  Mark Stapleton  Doug Amorese  Christophe Legendre  Heather E Cunliffe  Ann E McCullough  Barbara Pockaj  David Craig  John Carpten  Daniel Von Hoff  Christine Iacobuzio-Donahue  Michael T Barrett
Abstract:Formalin fixed paraffin embedded (FFPE) tissues are a vast resource of annotated clinical samples. As such, they represent highly desirable and informative materials for the application of high definition genomics for improved patient management and to advance the development of personalized therapeutics. However, a limitation of FFPE tissues is the variable quality of DNA extracted for analyses. Furthermore, admixtures of non-tumor and polyclonal neoplastic cell populations limit the number of biopsies that can be studied and make it difficult to define cancer genomes in patient samples. To exploit these valuable tissues we applied flow cytometry-based methods to isolate pure populations of tumor cell nuclei from FFPE tissues and developed a methodology compatible with oligonucleotide array CGH and whole exome sequencing analyses. These were used to profile a variety of tumors (breast, brain, bladder, ovarian and pancreas) including the genomes and exomes of matching fresh frozen and FFPE pancreatic adenocarcinoma samples.
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