This pilot study assesses the feasibility of genomic profiling on tissue obtained using a new core needle (EchoTip ProCore needle, Cook Medical Inc, Limerick Ireland). Methods: Four patients with pancreatic cancer underwent EUS guided
fine needle biopsy using a 19G or 22G needle to obtain a core specimen. Core specimens were extracted using Qiagen’s Qiasymnphony automated extractor using magnetic beads. A new low input material (200 ng) protocol Agilent Sureselect V4+UTR (71 Mb) was used for sequencing. DNA was quantitated using Picogreen & used for hybridization with the exome probes (WES) & sequenced with Hiseq 2000. Matched blood samples were collected for comparison analysis. Results: All four samples showed sufficient malignant cells for genomic analysis. The samples were sequenced to a mean exome coverage selleck of at least 95x (average of 115x), with an average of 92% of each exome covered by at least 20 reads (Table 1). Table 1 Exome Coverage and PCR Duplication Rates of Samples Sample Percent of Exome > 20X Coverage Average Coverage PCR Duplication Rate NG02CCK 92 110 6.60% TG02CCK 92 113 6.24% NG03AAB 91 108 6.71% TG03AAB 96 169 11.03% NG08LBY 91 99 6.15% TG08LBY 94 129 7.53% NG14OKS 90 95 5.41% TG14OKS 92 99 5.92% The samples showed known mutations in pancreatic cancer; Kras mutations (3 of 4), SMAD 4 mutations (1 of 4) and P53 mutations (2 of 4) (Table 2). Table 2 Mutations
Detected in Samples Pictilisib mw Mutations SAMPLE KRAS SMAD4 TP53 Note: All KRAS and TP53 mutations are known mutations. The SMAD4 mutation is frameshift and expected to be truncating. Conclusion: This is a first proof of concept study in performing genomics using a new core needle and low input genomic
sequencing system. Further studies with a larger sample size are required to show the feasibility of using this needle for genomic analysis. Key Word(s): 1. EUS; 2. pancreas carcinoma; 3. genomic; 4. core biopsy Presenting Author: KEISUKE TANIUCHI Additional Authors: MUTSUO FURIHATA, MCE公司 SHINJI IWASAKI, SHOGO SHIMIZU, TAKAHIRO SHIMIZU, MOTOAKI SAITO, TOSHIJI SAIBARA Corresponding Author: KEISUKE TANIUCHI Affiliations: Kochi Medical School, Kochi Medical School, Kochi Medical School, Kochi Medical School, Kochi Medical School, Kochi Medical School Objective: This study describes new and unique findings regarding the molecule RUVBL1 in pancreatic ductal adenocarcinoma (PDAC). Previous reports describe that RUVBL1 belongs to the family of AAA+ ATPases that participate in many cellular processes highly relevant to cancer. Methods: Immunoprecipitation and mass spectrometry were used to isolate and identify proteins that interact with RUVBL1. An in vitro actin polymerization assays and immunocytochemistry were used to examine the effects of RUVBL1 on the concentration of monomeric globular-actin (G-actin) and polymerization of filamentous actin (F-actin). Results: RUVBL1 accumulated in membrane protrusions and at the leading edges of PDAC cells.