bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase I study of weekly intraperitoneal paclitaxel combined with S-1 and oxaliplatin for gastric cancer with peritoneal metastasis.
Authors: Ishigami Hironori , Kaisaki Shoichi , Yamaguchi Hironori , Yamashita Hiroharu , Emoto Shigenobu , Kitayama Joji
Published: February 2017

146

Background: Intraperitoneal (IP) chemotherapy is a promising treatment option for gastric cancer with peritoneal metastasis. We previously verified the safety and efficacy of IP paclitaxel (PTX) combined with S-1 and intravenous PTX in phase I and phase II studies. S-1 plus oxaliplatin (SOX) demonstrated efficacy in a phase II study, and is regarded as a candidate for the next-generation standard regimen for gastric cancer. We designed a new regimen combining weekly IP PTX with SOX in order to maximize systemic effects as well as local effects in the peritoneal cavity. A dose-escalation study of IP PTX in combination with fixed doses of SOX was carried out to determine the maximum-tolerated dose (MTD) and recommended dose (RD).

Methods: PTX was administered intraperitoneally on days 1 and 8 with an initial dose of 20 mg/m2 (level 1), stepped up to 30 mg/m2 (level 2) or 40 mg/m2 (level 3) depending on observed toxicity. S-1 was administered orally at a dose of 80 mg/m2/day (b.i.d.) for 14 days followed by a 7-day rest. Oxaliplatin was administered intravenously at a dose of 100 mg/m2 on day 1. This treatment was repeated every 3 weeks. Toxicity was graded according to CTCAE v4.0. Dose-limiting toxicities (DLTs) were defined as grade 4 leukopenia, grade 3 febrile neutropenia, grade 3 thrombocytopenia, and grade 3 non-hematological toxicity. The MTD was defined as the dose level at which 2 or more of 3 or 6 patients developed DLTs during two courses of treatment. The RD was defined as one dose level under the MTD.

Results: A total of 12 gastric cancer patients with peritoneal metastasis were enrolled. No DLTs were observed at all dose levels. Neutropenia in one patient at dose level 3 was the only grade 3 toxicity observed. Grade 2/3 leukopenia, neutropenia and thrombocytopenia were observed only in 2 patients at dose level 3. Regarding grade 2 non-hematological toxicities, anorexia, fatigue and nausea were observed in 6, 4 and 2 patients, respectively, independent of dose levels. Consequently, the MTD was not reached, and the RD of IP PTX was determined to be 40 mg/m2 (level 3).

Conclusions: Combination chemotherapy of IP PTX with SOX was shown to be a safe regimen that should be further explored in clinical trials.
Detection of early-stage pancreatic ductal adenocarcinoma (PDAC): Sensitivity, specificity, and discriminatory properties of the serum-based PAM4-immunoassay.
Authors: Gold David V. , Gaedcke Jochen , Ghadimi B. Michael , Goggins Michael , Hruban Ralph H. , Liu Mengling , Newsome Guy , Goldenberg David M.
Published: February 2017

151

Background: We recently reported that a serum-based enzyme immunoassay employing the PAM4 antibody was able to correctly identify 82% of patients with known PDAC and, importantly, that this assay had promising sensitivity for detecting early-stage disease. We now extend these findings in a much larger patient population that includes over 600 sera from both malignant and benign diseases of the pancreas and surrounding tissues.

Methods: In a blinded analysis, sera from patients with confirmed PDAC (N=298), other cancers (N=99), benign disease of the pancreas (N=126), and healthy adults (N=79) were evaluated by enzyme immunoassay for concentration of PAM4-antigen levels.

Results: Overall sensitivity for detection of PDAC was 76%, with 64% of stage-1 patients testing positive and a higher sensitivity (85%) for advanced disease. For the most part, sera from patients with neuroendocrine tumors of the pancreas or cancers of other origin (squamous, GIST, etc.) did not have elevated levels of the PAM4-antigen. Approximately half of the patients with ampullary (48%) and extrahepatic biliary (50%) adenocarcinomas had positive levels of circulating PAM4-antigen. Of 126 patients diagnosed with benign conditions of the pancreas, only 24 (19%) were positive and, in particular, 18 of 80 (23%) patients with chronic pancreatitis (CP) were positive. ROC curve analysis demonstrated a statistically significant difference between the PDAC and CP groups (P < 0.0001), with an area under the curve of 0.84 ± 0.02 (95% CI: 0.79 – 0.89). The positive- and negative-likelihood ratios for differentiating PDAC from benign conditions of the pancreas were 4.00 and 0.30, respectively.

Conclusions: The PAM4-immunoassay detects nearly two-thirds of stage-1 PDAC patients, and does so with high discriminatory power with respect to benign pancreatic disease. Our results provide a rationale for longitudinal surveillance of patients considered at high-risk for PDAC (e.g., familial pancreatic cancer, new-onset diabetes, etc.) with the PAM4 assay. (Supported in part by NIH grants CA096924, CA120432 and CA62924, and the Turpin Foundation.)
Detection of resectable pancreatic cancer with SOMAmer proteomic technology.
Authors: Brand Randall , Buchholz Malte , Mehan Mike , Whitcomb David C. , Zeh Herbert , Moser Arthur J , Gress Thomas M , Williams Steve , dela Cruz Maria , Ostroff Rachel ,   European MolDiag-Paca Consortium
Published: February 2017

162

Background: To improve the current dismal 5-year survival rates for pancreatic adenocarcinoma (PC) it is essential to develop strategies focused on detection in high-risk populations. This study employed a unique SOMAmer proteomics platform to identify a biomarker panel that discriminates between pancreatic cancer and control patients. A secondary goal was to compare the panel with CA 19-9.

Methods: Plasma samples (total < 20 ul) were analyzed for 825-proteins using a SOMAmer proteomics platform on training set of 100 cases and 69 controls from a single institution. The control group contained individuals with GI symptoms, including acute and chronic pancreatitis, and normal controls. An independent blinded validation set from another center consisted of 43 PC patients and 47 controls. About two thirds (96/143) of the study cases were in resectable stages I-IIb. Biomarkers were identified by Random Forest backwards selection, using a false-discovery-rate corrected value of p<0.001. CA19-9 levels were available on a subset of the patients from the training set (87 PC and 29 controls). A CA 19-9 level of > 40 U/ml was defined as positive.

Results: In the training set, 37 markers were significantly different between case and controls. A 10 marker Random Forests classifier had an AUC of 0.91 for training and 0.90 for the independent validation set. Comparison of the SOMAmer results with CA 19-9 levels are shown in the table. There was 78% agreement between the two tests. One could maximize sensitivity with some sacrifice of specificity by combining either test positive (95% and 66%, respectively) or conversely maximize specificity (97%) with lower sensitivity of 77% if both tests must be positive.

Conclusions: 1) The 10 marker SOMAmer biomarker panel is promising for the detection of resectable pancreatic cancer with an AUC of 0.91 on a training set, which was confirmed on an independent validation set. 2) Preliminary data suggests that this test is complementary to CA 19-9 with improved combined sensitivity of up to 95%. 3) Studies to assess further applications of these panels, including for risk assessment in individuals with a family history of pancreatic cancer, are underway.

[Table: see text]
A population-based analysis of prognostic factors in patients with advanced hepatocellular carcinoma treated with sorafenib.
Authors: Smith Alan D. , Cheung Winson Y.
Published: February 2017

163

Background: Available clinical prognostic scoring systems for advanced hepatocellular carcinoma (HCC) were developed in the era of conventional chemotherapy. In 2008, the molecularly targeted agent sorafenib became the new standard of care for advanced HCC due to its survival benefit. The utility of these prognostic models in the setting of sorafenib is unclear. Our aims were to assess for new prognostic factors in patients treated with sorafenib and compare these with known prognostic systems.

Methods: All patients diagnosed with advanced HCC from 2008 to 2010 in British Columbia, Canada and treated with sorafenib at any 1 of 5 regional cancer centers were eligible. Based on the established Okuda, CLIP, Barcelona, and French staging systems, we collected baseline demographic and disease characteristics of patients prior to receipt of sorafenib. Multivariate logistic regression models were constructed to examine for associations between these clinical factors and overall survival.

Results: Of 183 patients identified, 152 were evaluable: median age was 63 years, 78% were men, average number of sorafenib treatment was 5.3 cycles, and median overall survival was 9.6 months. The prevalence of hepatitis B, hepatitis C, and alcohol-related liver disease were 32%, 15%, and 11%, respectively. Univariate analyses showed that poor performance status, presence of clinical ascites, as well as elevated serum AST, GGT, ALP, bilirubin and platelet levels were each associated with worse overall survival (all p<0.05). In multivariate analyses, however, none of these clinical factors continued to be independently predictive of outcome (all p>0.05).

Conclusions: Traditional clinical prognostic factors developed in the era of conventional chemotherapy do not appear to have the same prognostic utility in this contemporary Western cohort of advanced HCC patients treated with sorafenib. This observation underscores the need to identify molecular biomarkers that provide better prognostic information.
Blinded independent central response assessment using RECIST, mRECIST, and CHOI criteria in patients treated with sorafenib for advanced hepatocellular carcinoma (HCC).
Authors: Bouattour Mohamed , Wassermann Johanna , Bruno Onorina , Dreyer Chantal , Larroque Beatrice , Castera Laurent , Vilgrain Valérie , Belghiti Jacques , Raymond Eric , Faivre Sandrine J.
Published: February 2017

172

Background: Despite survival benefit in patients with advanced HCC, sorafenib has been infrequently associated with changes in tumors dimensions, challenging standard RECIST criteria. Experts have suggested new response criteria based on changes in tumor vascularization and density (mRECIST and Choi). Aim is to evaluate response in patients treated with sorafenib using three different methods and their correlation with overall survival (OS)

Methods: Radiological evaluations were performed from consecutive patients treated with sorafenib from January 2007 to February 2009. Patients were considered evaluable if CT scans were performed within 6 weeks before sorafenib and in the three months following treatment initiation at the first evaluation. A radiologist blinded for clinical data performed tumor evaluations according to RECIST, mRECIST, and Choi criteria. Best response was correlated with OS

Results: Among 82 patients, 60 were evaluable (median age 63, male/female 52/8). The majority of patients were Child A (80%). Tumor staging showed BCLC stage B and C in 33% and 67%, respectively. The median duration of treatment with sorafenib was 6.6 months (range 1.2-16.5). Tumor evaluations by CT scan were performed in a median of 2.1 months (range 1.5-3.2). For the entire cohort, median OS was 9.9 months, and survival rates at 6, 12, and 24 months were 69%, 39% and 22%, respectively. The table summarizes the best responses measured using RECIST, mRECIST, and Choi criteria.

Conclusions: Better OS was observed in patients with complete/partial response than in patients with stable disease and progression, regardless the type of radiological criteria. However, mRECIST and Choi criteria identified more adequately than RECIST subgroups of patients with complete/partial response who also experienced longer survival

[Table: see text]
Baseline demographics of patients with neuroendocrine tumors presenting to seven National Comprehensive Cancer Network (NCCN) institutions: Development of a multi-institutional outcomes database.
Authors: Choti Michael A. , Mayorga Margaret A. , Bobiak Sarah , Kulke Matthew , Yao James C. , Bergsland Emily K. , Nakakura Eric K. , Bloomston Mark , Benson Al Bowen , Shah Manisha H. , Strosberg Jonathan R.
Published: February 2017

187

Background: Diagnostic strategies, management paradigms, and clinical outcomes of patients with neuroendocrine tumors (NETs) are diverse and poorly characterized. The National Comprehensive Cancer Network (NCCN) created a comprehensive longitudinal database to characterize patients treated for NETs at seven participating institutions. Preliminary results from the database are reported.

Methods: Member IRB approval was obtained to identify patients at least 18 years of age presenting to each of seven NCCN institutions between 2004 and 2007 with pathologically confirmed newly or previously diagnosed NETs via hospital medical records. Eligible patients included those with carcinoid (any site); goblet cell or adenocarcinoid; composite carcinoid; poorly differentiated gastrointestinal small cell tumor; pancreatic NET; NET of unknown primary site; pheochromocytoma; and paraganglioma. Baseline demographic characteristics were summarized for this analysis.

Results: Among the 2,542 patients identified with a NET diagnosis, patients most frequently presented with carcinoid tumor (51%), pancreatic NET (27%) and NET of unknown primary site (8%). Median age at diagnosis was 55 (SD=13) and median age at first presentation to the NCCN was 57 (SD=13). Fifty-three percent of patients were female. Most (88%) were Caucasian and 8% were African American. Forty percent of patients were diagnosed with NET before presenting to the NCCN. Among these, the median time between initial NET diagnosis and presentation to the NCCN was 2 years (SD=6). Significant differences in provider specialty referral patterns were observed between institutions.

Conclusions: The baseline demographic characteristics of NET patients in this new database are consistent with those previously reported in population-based registries. The NCCN database will provide a valuable resource for further exploration of patterns of diagnosis, treatment, consistency with established guidelines, as well as clinical outcomes in patients with this condition. Existing differences among institutions in referral patterns will be further explored.
Symptom changes that predict disease control by systemic chemotherapy in patients with advanced pancreatic cancer.
Authors: Kuwahara Akiko , Mitsunaga Shuichi , Ohno Izumi , Shimizu Satoshi , Takahashi Hideaki , Okuyama Hiroyuki , Okusaka Takuji , Ueno Hideki , Morizane Chigusa , Kondo Shunsuke , Ikeda Masafumi
Published: February 2017

195

Background: Alleviation of symptoms is related to good tumor control in patients undergoing systemic chemotherapy for pancreatic cancer (PC). The predictive value of symptom change has not been fully understood. The aim of this study was to identify symptom changes that predicted disease control by systemic chemotherapy in PC patients.

Methods: Patients with unresectable PC who had not received any anti-cancer therapy for PC were eligible for inclusion in this study. Patients with obvious infectious conditions were excluded. Symptoms were scored from 0 to 10 using the Japanese version of the MD Anderson Symptom Inventory before the start of chemotherapy and one month later. The response of the tumor was evaluated using Response Evaluation Criteria in Solid Tumors (RECIST) vs. 1.0. Disease control was defined as CR, PR, or SD. An attempt was made to identify symptom changes that predicted disease control, and their impact on progression-free survival (PFS) was assessed.

Results: The symptoms of 87 patients (male/female: 46/41, Karnofsky performance status (KPS): 100/90/80/70-50: 32/29/17/9, median age: 66 years) were prospectively assessed. Gemcitabine monotherapy (GEM), a GEM-based regimen, and S-1 monotherapy were performed in 42, 41, and 4 patients, respectively. Disease control was observed in 31 patients. The pain scores (P=0.047) and sleep disturbance scores (P=0.052) one month later were lower than before treatment in patients with disease control. Patients without deterioration of pain or sleep disturbance had a higher frequency of disease control (P=0.098 and P=0.004, respectively) and a longer PFS (P=0.021 and P<0.001, respectively) than patients whose pain or sleep disturbance became more severe.

Conclusions: Improvements in pain and sleep disturbance scores at one month predict disease control by systemic chemotherapy in patients with advanced PC.
Transforming growth factor (TGF) beta pathway and clinical outcome of pancreatic cancer.
Authors: Javle Milind M. , Tan Dongfeng , Li Yanan , Kar Siddhartha P. , Baladandayuthapani Veera , Dong Xiaoqun , Weatherly Jacqueline , Krishnan Sunil , Huang Tzu-chuan Jane , Fogelman David R. , Abbruzzese James L. , Wolff Robert A. , Li Donghui
Published: February 2017

201

Background: TGF-beta acts both as a tumor suppressor or tumor promoter depending upon cellular context and stage of tumor progression. TGF-β exerts its effects through the TGF-beta-R1/ R2 receptors and the SMAD transcription regulators. We investigated the prognostic value of TGFβ signaling biomarkers in pancreatic cancer.

Methods: We measured plasma TGF-beta-1 level using Meso Scale Discovery Multi-array Human TGF-beta-1 Assay in 643 pancreatic cancer patients. TGF-beta-R2 and SMAD4 protein expression were measured in 86 biopsies using immunohistochemisty (IHC). IHC intensity was scored 0-3. Percentage IHC-positive cancer cells were 0-3 (0: 0%+, 1: 1-10%+, 2: 10-25%+, 3: >25%+). Final IHC score was measured as intensity x percentage. We also genotyped 3 SNPs of TGFB1 gene using the Taqman assay. Kaplan-Meier and log-rank tests were used to analyze overall survival (OS) by TGFβ1 level. Multivariate Cox proportional hazards models with relevant clinical covariates were used to assess the relationship between IHC score and OS.

Results: In patients with locally advanced and metastatic disease (n = 355), there was a significant association of plasma TGF-beta-1 with overall survival (OS). OS of the top quartile of TGF-beta-1 levels (>19.05 ng/mL) vs. those with low level was 27.7 weeks vs. 40 weeks, respectively [log-rank p = 0.0125, adjusted for baseline CA 19-9 and performance status (PS)]. No significant association between plasma TGF-beta-1 and OS was noted in surgical patients (n=288). In the multivariate Cox model, after adjusting for baseline stage, CA 19-9, PS, age and TGF-beta-R2 expression, complete loss of SMAD4 expression (IHC score 0) was significantly associated with lower OS (HR: 1.85, 95% CI: 1.06-3.23; p = 0.03). Progressive disease on gemcitabine-based therapy was more likely in SMAD4 IHC 0 group as compared with higher score (46.5% vs. 38.1% progressed; chi square p = 0.069). TGFB1 -1346T>C CT/TT genotype correlated with shorter OS [HR (95% CI): 1.21 (1.02-1.45), p=0.03] after adjusting for clinical variables.

Conclusions: This large retrospective study suggests an important stage-dependant role of the TGF-beta pathway in pancreatic cancer. Targeting this pathway in advanced pancreatic cancer may have therapeutic relevance.
Staging of advanced hepatocellular carcinoma patients in the targeted therapy era.
Authors: Shah Neeraj Nailesh , Hassan Manal , Xiao Lianchun , Abbruzzese James L. , Morris Jeffrey , Kaseb Ahmed Omar
Published: February 2017

209

Background: Several hepatocellular carcinoma (HCC) staging systems are currently available. However, they were all developed before the targeted therapy era prevailed in the last decade which has changed the natural history of the disease. Our study goal was to test the performance of different HCC staging systems in patients with HCC who were treated at our institution during the last decade.

Methods: We prospectively enrolled 438 patients from early 2000 to late 2009. Baseline clinicopathologic parameters and staging were available, including the TNM, Cancer of the Liver Italian Program (CLIP), Barcelona Clinic Liver Cancer (BCLC), Okuda, and Chinese University Prognostic Index (CUPI). We performed survival and cox-regression analyses, and compared the staging systems’ predictive ability using Harrell's C-index. Finally, we performed a subgroup analysis of 3 independent cohorts based on whether or not they received sorafenib, whether or not they had hepatitis, and whether or not they had cirrhosis.

Results: The overall survival was 13.9 months. Overall, CLIP score was the most predictive staging system with a C-index of 0.71. 187 patients were treated with targeted therapies and 138 were treated with sorafenib after it was approved in 2007. CLIP score was the most predictive staging system with a C-index of 0.71 in the no sorafenib group, and 0.74 in the sorafenib group. In hepatitis patients, CLIP topped amongst all staging systems with a C-index of 0.75, and in patients without hepatitis, despite all staging systems having a poor predictive ability, CLIP score still had the highest C-index of 0.67. Similarly, CLIP score had the best predictive ability in patients with and without pre-existing liver cirrhosis, with C-indices of 0.73 and 0.68 respectively. There was no statistically significant interaction between CLIP score and hepatitis status, and CLIP score and liver cirrhosis. Thus, overall the CLIP score was the best predictive system in all cohorts.

Conclusions: Our results suggest that the CLIP score has the highest stratification ability in our advanced HCC patient population, including several subgroups. Our study confirms the utility of the CLIP score to stratify advanced HCC patients in clinical trials.
An integrated cross-species approach for the discovery of serum-based diagnostic biomarkers of PDAC using a genetically engineered murine model (GEMM).
Authors: Skoulidis Ferdinandos , Schiess Ralph , Aebersold Ruedi , Venkitaraman Ashok R.
Published: February 2017

218

Background: The development of sensitive and specific biomarkers for the early diagnosis of human pancreatic ductal adenocarcinoma (PDAC) represents a major unmet clinical need. GEMMs that faithfully recapitulate the clinical, histological and molecular features of PDAC and capture its genomic complexity (including that triggered by a truncating germline Brca2 mutation) coupled with state of the art proteomic technologies can overcome some of the challenges associated with the analysis of a heterogeneous human disease. We have conducted a large scale screen for differentially expressed glycoproteins in pancreatic tissue and serum from mice with PDAC, pancreatic intra-epithelial neoplasia (PanIN) and matched healthy controls, using the well validated KPCB model of PDAC.

Methods: Tryptic digests of protein extracts from snap frozen PDAC, pre-malignant and normal pancreata were subjected to solid-phase extraction of N-glycopeptides and the samples were analysed on a LTQ Orbitrap XL mass spectrometer. LC-MS feature maps were compared employing SuperHirn software and used for the label-free quantification of corresponding proteins. Primary mPDAC cell lines and their supernatants were also analysed. Selected candidate proteins were then quantified in mouse serum by SRM-based targeted proteomics.

Results: 146 glycoproteins were identified as overexpressed by >2 fold (p<0.05, Student’s t test) in PDAC compared to normal pancreas and 60 were prioritised for targeted identification in mouse serum. 12 proteins were significantly upregulated (p<0.01, Mann-Whitney U test ) in the serum of tumour-bearing mice with PDAC compared to controls.

Conclusions: Using a state of the art GEMM of PDAC, we have successfully applied a two-stage strategy for the tissue-based identification followed by quantification in murine serum of a panel of glycoproteins that are consistently over-expressed during mPDAC development. Validation in serum collections from patients with PDAC, chronic pancreatitis and healthy controls is underway and updated results will be presented at the conference.
Association of KRAS G12D mutation with outcome in patients with pancreatic cancer after surgery.
Authors: Mikayama Yo , Morinaga Soichiro , Watanabe Takuo , Numata Masakatsu , Tamagawa Hiroshi , Yamamoto Naoto , Shiozawa Manabu , Akaike Makoto , Yoichi Kameda , Matsukuma Shoichi , Miyagi Yohei
Published: February 2017

221

Background: The frequency of KRAS gene mutations in pancreatic adenocarcinoma is high and most mutations occur in codon 12 and 13. However, the association of KRAS mutation status with clinical outcome of pancreatic cancer patients after surgery is not clearly understood. The aim of present study was to clarify whether presence of mutation in KRAS is of prognostic significance, and to determine whether the different type of KRAS mutation have different impact on clinical outcome in pancreatic cancer.

Methods: A total of 62 pancreatic adenocarcinoma tissues from patients underwent curative surgery were analyzed for KRAS mutation. DNA was extracted from the tissue microarray. The domain containing the KRAS gene codons 12 and 13 was amplified by PCR, and KRAS mutation was detected by loop hybrid mobility shift assay (LH-MSA).

Results: KRAS mutation was detected in 41 of 62 tumors (66.1%). Mutations of KRAS codon 12 and codon 13 were detected in 37 of 62 tumors (59.7%) and in 4 of 62 tumors (6.5%), respectively. G12D mutation were detected in 35.5% (22/62) and G12V mutation in 19.4% (12/62) of all. The patients with KRAS mutations had a significantly shorter overall survival (OS) (log rank, p=0.0375) and disease free survival (DFS) (p=0.0175). When the type of mutation were evaluated, G12D mutation was significantly associated with both shorter OS (p=0.0410) and DFS (p=0.0214), whereas G12V mutation was associated with neither OS (p=0.6727) nor DFS (p=0.9854).

Conclusions: The presence of KRAS mutation was significantly associated with poor outcome of pancreatic cancer patients after surgery. A specific mutation G12D demonstrated significant association with shorter OS and DFS in pancreatic cancer. The different type of KRAS mutation might have different impact on clinical outcome in pancreatic cancer.
Comparative analysis of myeloid-derived suppressor cell (MDSC) subsets in patients with gastrointestinal (GI) malignancies.
Authors: Duffy Austin G. , Zhao Fei , Fioraventi Suzanne , Greten Tim
Published: February 2017

228

Background: Immune-based therapies have begun to demonstrate efficacy in GI oncology. However immune-suppressor mechanisms have been described which interfere with host anti-tumor immunity. Myeloid derived suppressor cells (MDSCs) constitute a heterogenous population known to be increased in patients with GI cancer. MDSCs inhibit the host anti-tumor immune response, limiting the effectiveness of both immune and non-immune treatments. Characterization of MDSCs in humans has been difficult with heterogeneous phenotype classification. Differences in processing are also potentially important. The aim of this study was to perform a comparative analysis of MDSC subsets in patients with GI cancer.

Methods: Fluorescence activated cell sorting (FACS) analysis was performed on PBMC and whole blood (WB) lysates from both healthy adults and patients with advanced hepatocellular, colorectal and pancreatic cancer. Antibodies used for subtype characterization: anti-CD14, anti-CD15, anti-HLA-DR, anti-CD33, anti-CD11b. Functional studies performed to confirm MDSC status included thymidine proliferation assay on sorted cell populations.

Results: Samples were obtained from 20 patients with advanced GI cancer and healthy volunteers and processed accordingly. Based on published data the following MDSC subtypes were analyzed: A) CD14+, HLA-DR−/low; B) CD15+,CD14-CD33+CD11b+.The frequency of each subset is shown in the table below. Functional studies will also be presented.

Conclusions: Characterization of MDSCs in humans has been difficult with heterogeneous and often contradictory phenotype classification. In addition, there is considerable variability depending on which component of the blood is analyzed and how the sample is processed. This study confirms the increase in both MDSC populations in patients with GI cancer.

[Table: see text]
Sorafenib-induced loss of polarity of zymogen granules in pancreatic acinar cells.
Authors: Ito Tatsuo , Doi Ryuichiro , Uemoto Shinji
Published: February 2017

236

Background: Sorafenib is an oral multi-kinase inhibitor which is regarded as a key drug for HCC and RCC. It has been unexpectedly found that the compound causes an increase of serum pancreatic enzyme levels without clinically recognized pancreatitis. The reason for this event is not well understood yet. The aim of this study was to clarify the mechanisms involved in this phenomenon.

Methods: Eight-week old BALB/cA male mice were used in in vivo studies. Sorafenib tosylate was administered per os once daily at a dose of 150 mg/kg body weight. Control mice were given vehicle alone. Mice were sacrificed 24 hr after 1-, 2-, 3- and 7-day administration of the compound, and blood samples and pancreatic tissue samples were obtained (n=5 for each group). The tissue samples were used for hematoxylin and eosin (HE) staining, immunohistochemistry, electron microscopy (EM), western blot and RT-quantitative PCR studies.

Results: Serum amylase levels were elevated after sorafenib administration. The amylase level hit the peak after 2-day administration, and then gradually decreased. By HE staining, the control group without sorafenib showed a basophilic stained area in the baso-lateral site of the acinar cells. In contrast, the acinar cytoplasm after 2-day administration of sorafenib was totally eosinophilic. The typical findings of acute pancreatitis were not seen in the both group. By EM examination, zymogen granules (ZGs) of the sorafenib group spread into basal site of the acinar cells. ZGs mounted up on both of apical and baso-lateral plasma membrane and showed exocytosis. The levels of amylase mRNA were not elevated by sorafenib. In addition the expression of N-ethylmaleimidesensitive factor attachment protein receptor (SNARE) proteins was not changed.

Conclusions: The results suggest that the amount of acinar amylase production was not changed but the distribution of ZGs was altered by sorafenib. Sorafenib seemed to cause temporary loss of polarity of ZGs secretion in acinar cells by blocking apical exocytosis. Acute pancreatitis was not evident; thus the current model was not similar to the pancreatitis model caused by the supra-maximal secretagogue stimulation which blocks the apical exocytosis.
Preclinical study of MEK1/2 inhibitor AZD6244 combined with gemcitabine for treatment of biliary cancer.
Authors: Xu Junyao , Knox Jennifer J. , Tsao Ming Sound , Chen Eric Xueyu , Cao Pinjiang , Sue Chow , Serra Stefano , Hedley David W
Published: February 2017

240

Background: MEK1/2 is an integral component of the Ras/Raf/MEK/ERK signaling pathway, implicated in uncontrolled cell proliferation and cell survival, a key hallmark of cancer. AZD6244, a novel inhibitor of MEK1/2, is currently completing Phase II clinical trials in biliary cancer, with modest antitumor activity observed as monotherapy. Gemcitabine is a cytotoxic drug commonly used in biliary cancer therapy but many patients showed early resistance. In this preclinical study, we investigated the sequence-dependent antitumor effects of AZD6244 combined with gemcitabine in biliary cancer models.

Methods: Two biliary cancer cell lines (EGI-1 and TFK-1) were used. In vitro the effects of single drug or three combination protocols(concurrently; AZD6244 followed by GEM or Gem followed by AZD6244) on cell proliferation, DNA synthesis, and cell cycle distribution were evaluated by MTS, clonogenic assay, EdU uptake and flow cytometry. Drug interactions were analyzed by Chou-Talaly method. In vivo, 4 tumor models subcutaneously xenografted in SCID mice from the two cell lines and 2 human patients were set up to compare the therapeutic effects of different sequence-scheduled combinations.

Results: AZD6244 caused G1-S cell cycle arrest in biliary cancer cells in vitro and in vivo, and this effect is correlated with the MEK/ERK signaling pathway blocking. Synchronized progression of the population through S phase were observed in 15h after removal of AZD6244 in cell culture or 48h after final dose of acute AZD6244 treatment in vivo. Antagonistic or additive effects was observed in vitro when combination were given as concurrently(CI=2.03~2.46) or Gem followed by AZD6244(CI=1.34~1.78). In contrast, a synergistic antiproliferative activity was obtained when AZD6244 was given first followed by a drug-free interval before Gem treatment (CI=0.53~0.69). In vivo, the best therapeutic effects were obtained with the sequence of AZD6244 followed by Gem, compared with concurrent or reverse sequence.

Conclusions: This study provides a sound rationale for a Phase II trial of a potentially synergistic sequence of MEK inhibitor AZD6244 followed by gemcitabine in patients with advanced biliary cancer.
Effect of NVP-BEZ235, a dual PI3K/mTOR inhibitor, on chemotherapy and antiangiogenic response in pancreatic cancer.
Authors: Ostapoff Katherine T. , Awasthi Niranjan , Yen Peter L. , Zhang Changhua , Schwarz Margaret A. , Brekken Rolf A. , Schwarz Roderich
Published: February 2017

243

Background: The phosphatidylinositol-3-kinase (PI3K)/AKT and mammalian target of rapamycin (mTOR) signaling pathway dysregulation is a prominent feature of pancreatic ductal adenocarcinoma (PDAC). Gemcitabine (GEM), a standard systemic treatment for PDAC, has limited clinical benefits. The present study investigated the effects of NVP-BEZ235 (BEZ235), a novel dual PI3K/mTOR inhibitor, in combination with gemcitabine and endothelial monocyte activating polypeptide II (EMAP) in experimental PDAC.

Methods: Protein expression and cell proliferation were analyzed by Western blotting and WST-1 assay. Animal experiments were performed in murine xenografts.

Results: BEZ235 inhibited phospho-AKT (Ser473) and phospho-mTOR (Ser2448) expression in PDAC (AsPC-1), endothelial (HUVECs) and fibroblast (WI-38) cells. NVP-BEZ235 also caused a significant dephosphorylation of downstream mTORC1 target proteins phospho-p70 S6K (Thr389) and phospho-4E-BP1 (Thr37/46). In vitro 72-hour proliferation of four PDAC cell lines was significantly inhibited by BEZ235. Additive effects on proliferation inhibition were observed in the BEZ235 and GEM combination in PDAC cells and in combination of BEZ235 or EMAP with gemcitabine in HUVECs and WI-38 cells. BEZ235, alone or in combination with GEM and EMAP, induced apoptosis in AsPC-1, HUVECs and WI-38 cells as observed by increased expression of cleaved poly (ADP-ribose) polymerase-1 (PARP-1) and caspase-3 proteins. PDAC in vivo therapy demonstrated that compared to controls (median survival: 16 days), animal survival increased after BEZ235 and EMAP therapy alone (both 21 days) and GEM monotherapy (28 days). Further increases in survival occurred in combination therapy groups BEZ235+GEM (30 days, p=0.007), BEZ235+EMAP (27 days, p=0.02), GEM+EMAP (31 days, p=0.001) and BEZ235+GEM +EMAP (33 days, p=0.004).

Conclusions: BEZ235 has experimental PDAC antitumor activity in vitro and in vivo that can be enhanced in combination with cytotoxic (GEM) and antiendothelial (EMAP) agents. These findings demonstrate advantages of combination therapy strategies targeting multiple pathways in pancreatic cancer treatment.
Significance of mortalin expression in gastric cancer with normal p53.
Authors: Ando Koji , Oki Eiji , Yan Zhao , Kitao Hiroyuki , Saeki Hiroshi , Ohga Takefumi , Kakeji Yoshihiro , Tsujitani Shunichi , Kohnoe Shunji , Maehara Yoshihiko
Published: February 2017

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Background: Gastric cancer still has the highest morbidity rate and the second highest mortality rate. As chemotherapy confers only a minimal survival advantage, the prognosis of patients with advanced or recurrent gastric cancer remains poor. Studying the mechanisms and underlying molecules that drives gastric cancer malignant, could contribute to finding a remedy for gastric cancer. Mortalin is a heat non-inducible member of the heat shock protein 70 family. Mortalin binds to p53 and prevents p53 from entering the nucleus, as well as cell stress. To understand the significance of mortalin in gastric cancer, we investigated the expression of mortalin and p53.

Methods: The expression of mortalin and p53 were examined by immunohistochemical staining of 182 clinical samples of gastric cancer.

Results: Mortalin-positive and aberrant-p53-positive samples were found in 75.2% and 62.6% of cases, respectively. Mortalin-positive tumors were deeper in invasion and had more lymph node and liver metastases compared with mortalin-negative tumors (p < 0.01, p < 0.05, respectively). Mortalin-positive tumors had worse prognosis compared with mortalin-negative tumors (p = 0.035). Moreover, in tumors with normal p53 expression, mortalin-positive tumors had especially worse prognosis compared with mortalin-negative tumors (p = 0.013). With multivariate analysis, mortalin expression appeared to be an independent prognostic factor in gastric cancer with expression of normal p53 (p = 0.0174).

Conclusions: Mortalin has a great impact on gastric cancer with normal p53. Therefore, mortalin is a target molecule for treatment of gastric cancer, as well as a promising prognostic factor, especially in tumors with normal p53.
Percutaneous irreversible electroporation (IRE) in the treatment of hepatocellular carcinoma (HCC) and metastatic colorectal cancer (mCRC) to the liver: A retrospective analysis of the University of Miami experience.
Authors: Loaiza-Bonilla Arturo , Barbery Katuska J , Perez-Rojas Evelyn , Froud Tatiana , Yrizarry Jose M , Akunyili Ikechukwu Immanuel , Rocha Lima Caio Max S. , Feun Lynn G. , Hosein Peter Joel , Narayanan Govindarajan
Published: February 2017

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Background: Percutaneous ablation and transarterial chemo- or radioembolization are commonly used in the treatment of HCC and mCRC to the liver. IRE using the Nanoknif is more versatile than other ablative modalities (such as RFA) in that tumors abutting vascular structures can be treated with IRE without compromise of the vessels or concern for the heat sink effect of nearby blood flow.

Methods: We examined the records of patients (pts) referred for IRE for HCC and mCRC. The procedures were all done percutaneously under general anesthesia using a standard protocol. The primary endpoint was progression-free survival (PFS). Responses were assessed using the modified RECIST criteria.

Results: Between 1/2010 and 8/2011, 49 pts underwent percutaneous ablation of unresectable HCC and mCRC liver tumors using IRE – 33 with HCC and 16 with mCRC. A total of 76 lesions were treated in 62 sessions; the median number of lesions treated per patient was 1 (range 1-4) and the median tumor size treated was 2.1cm (range 0.8-6). After IRE, 20 pts (41%) had a complete response (CR), 19 (39%) had a partial response and 10 (20%) had stable disease as their best response. The Kaplan-Meier estimated median PFS was 11.3 months (95% CI 9.6-12.9) for all pts, 11.6 months (95% CI 10.2-12.9) for HCC pts, and 10.4 months (95% CI 5.4-15.4) for mCRC patients. The one-year PFS was significantly higher for pts achieving a CR compared to those who did not achieve a CR (75% versus 59%, log rank p = 0.05). The number of liver lesions at baseline and size of the treated lesions were not associated with any differences in survival. The IRE was complicated in 6 pts (12%) by pneumothorax (2), pleural effusion (2), and atrial flutter/fibrillation during anesthesia (2). All pts recovered fully from these complications. One pt died within 1 month of the IRE due to disease progression.

Conclusions: IRE of liver tumors is safe. The PFS rates for pretreated mCRC and unresectable HCC are promising. A complete lack of enhancement of the treated lesion on the post-IRE CT scan appears to be associated with longer survival.
A phase I trial of isolated hepatic perfusion (IHP) using 5-FU and oxaliplatin in patients with unresectable isolated liver metastases (ILM) from colorectal cancer (CRC).
Authors: Magge Deepa , Zureikat Amer H , Bartlett David L. , Holtzman Matthew Peter , Choudry Haroon , Beumer Jan H. , Holleran Julianne L. , Strychor S. , Pingpank James F. , Zeh Herbert
Published: February 2017

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Background: IHP is a proven approach for regional delivery of chemotherapy in patients with unresectable ILM. This study sought to determine safety and MTD of administering 5FU in combination with fixed dose oxaliplatin via IHP.

Methods: Prospective Phase I dose escalation with standard 3x3 dosing. Subjects with unresectable ILM from CRC scheduled to receive an HAI pump were eligible. IHP employed fixed dose oxaliplatin (previously established)with escalating doses of 5FU. 1 endpoint was to determine MTD for this combination. 2 endpoints were response to IHP alone, PFS, and OS for IHP + HAI-FUDR. Systemic and IHP plasma PK of 5FU, anabolites, total, and free platinum were determined by validated assays.

Results: Between Aug 2007 - Mar 2011, 11 subjects were enrolled. All patients received at least one line of pre-IHP systemic chemotherapy. There were 4 Grade 3 SAE (36.3%) and no grade 4 events. 2 DLTs occurred in the second dose cohort of 300mg/m2. Dose escalation was terminated and 200mg/m2 5FU was determined to be the MTD. There was 1 DLT in the dose de-escalating phase of 200mg/m2. At first follow-up, 9 pts (82%) had a partial response, while 2 (18%) had stable disease. 64% of pts had a >50% decrease in CEA level. Actuarial 1 and 2 year survival was 100% and 75% respectively, with median follow-up of 23 mos. IHP exposures (AUC0-60min) were 10.9±4.5 µgPt·h/mL (platinum), 49.3±30.7 µg·h/mL 5FU (DL1) and 70.5±35.5 µg·h/mL 5FU (DL2). Free platinum represented 82±14% of total platinum. Systemic exposure (AUC0-inf) relative to IHP exposure was negligible for both total platinum (1.1±1.5%) and 5FU (0.09±0.10%). IHP exposure to metabolites relative to 5FU was 4.9±2.5% for FUrd and 0.23±0.14% for FdUrd, respectively.

Conclusions: MTD for IHP with 5FU and oxaliplatin is 200 mg/m2 and 40mg/m2 respectively. Systemic exposure to the agents was minimal. The response and survival observed in this dose escalation study warrants assessment in a larger phase II trial.

[Table: see text]
Using the decline in CA19-9 after the start of chemotherapy to predict survival in patients with inoperable bile duct cancer.
Authors: Grunnet Mie , Baeksgaard Lene , Laursen Inga , Lassen Ulrik Niels , Sorensen Morten
Published: February 2017

287

Background: CA 19-9 has been approved by the FDA as a tumour marker in pancreatic cancer. The prognostic value of CA19-9 in cholangiocarcinoma is unclear.The aim of the study was to determine if an early CA 19-9 response after the first cycle of chemotherapy is predictive of survival in patients with inoperable bile duct cancer.

Methods: CA 19-9 response was defined as a 20% decline in CA 19-9 compared to baseline after four weeks of chemotherapy, in patients with a baseline CA 19-9 level above 1.5 times the upper limit of normal (ULN). Patients were identified retrospectively database at a single institution. Chemotherapy used was gemcitabine day 1 and 15 (1g/m2), oxaliplatin day 1 and 15 (60 mg/m2) and capecitabine (1g/m2 BID) days 1 to 7 and days 15-21 for six cycles. Patients with a baseline CA 19-9 above 1.5 the ULN and at least one CA 19-9 value measured four weeks after the start of chemotherapy were included in the study. Survival in CA 19-9 responders was compared to non-responders by use of the log rank test. A Cox regression analysis was performed using performance status, gender, disease extent and location, need for endoluminal stenting, baseline CA 19-9, and CA 19-9 response as co-variables.

Results: 212 patients were registered from Feb 2004 until Nov 2010. Among these, 170 were treated with chemotherapy for cholangiocarcinoma and 131 had a baseline CA 19-9 value and at least one follow-up CA 19-9. Patients with baseline CA 19-9 less than 1.5 times the ULN (n=45) or inconclusive measurements (n=7) were excluded leaving 79 patients in the study. Fifty per cent of patients had a CA 19-9 response to chemotherapy. The median survival in the CA 19-9 response group was 11.7 months (95%CI: 7.5 – 16.0) vs. 8.2 months (95% CI: 6.4 – 9.8) in the non-response group. CA 19-9 response (HR: 0.535 (95%CI; 0.331-0.866), p=0.01) and disease extent (metastasis vs. no metastasis) (HR: 0.604 (95%CI; 0.369-0.99) p=0.046) were independent predictors of survival in a Cox model.

Conclusions: CA19-9 response is useful as predictor of survival in patients with inoperable cholangiocarcinoma undergoing chemotherapy.
Sorafenib (S) in patients (pts) with advanced hepatocellular carcinoma (HCC): Safety and efficacy in Child-Pugh (CP) class A and B patients.
Authors: Pressiani Tiziana , Boni Corrado , Rimassa Lorenza , Labianca Roberto , Fagiuoli Stefano , Ardizzoni Andrea , Foa Paolo , Cortesi Enrico , Giordano Laura , Santoro Armando
Published: February 2017

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Background: S is the first systemic agent that has been shown to prolong survival in pts with CP A advanced HCC. However, its safety and efficacy have not been extensively evaluated in pts with CP B cirrhosis.

Methods: We performed a descriptive analysis on pts with histologically documented advanced HCC and CP A/B cirrhosis, enrolled in a multicenter phase II randomized, open-label trial (data reported elsewhere), in order to assess the feasibility and efficacy of treatment with S in CP B pts. Written informed consent was obtained from all pts.

Results: From April 2007 to July 2008, 297 pts were prospectively treated with S 400 mg bid, 234 (78.8%) CP A, 63 (21.2%) CP B. 232 pts were male (76%), median age was 68.3 yrs (range 19.8-89.2 yrs), 217 pts had no extra hepatic disease (73.1%). The two subgroups, according to CP class A or B, were homogeneous for all considered parameters. Median treatment duration was >3 months (mos) for 59.4% of CP A pts and for 27% of CP B pts (p <0.001). Median PFS for the total population was 3.9 mos, 4.3 mos for CP A pts and 2.1 mos for CP B pts (p<0.001). Median OS was 10 mos for CP A pts and 3.8 mos for CP B pts (p<0.001). Adverse events (all grades) were similar in type and incidence for CP A and B pts, with fatigue, stomatitis, diarrhea and weight loss having the highest incidence (44%, 41%, 38% and 38%, respectively). Specifically, no differences in terms of grade 3-4 events have been documented between the two groups.

Conclusions: This study supports the feasibility of S also in advanced HCC pts with CP B status. Tolerability data suggest that pts with CP B status might potentially be treated safely with S for its potential survival benefits. Further prospective trials, specifically designed to investigate S in CP B pts, are eagerly advocated.