bluuebunny/crossref_metadata_2025 huggingface dataset:
Updated analysis: Observational cohort study of first-line bevacizumab combined with chemotherapy in metastatic colorectal cancer (HGCSG0802)—Sub-group analysis by KRAS Exon2 status.
Authors: Nakamura Michio , Yuki Satoshi , Sawada Kentaro , Oba Ayane , Sato Atsushi , Kato Takashi , Meguro Takashi , Ehira Nobuyuki , Tateyama Miki , Hatanaka Kazuteru , Eto Kazunori , Okuda Hiroyuki , Kobayashi Yoshimitsu , Muto Osamu , Abe Masakazu , Kato Kanji , Kudo Mineo , Miyashita Kencho , Sakata Yuh , Komatsu Yoshito
Published: February 2017

522

Background: A few reports have shown the efficacy of bevacizumab (BV) independent of the KRAS Exon2 mutational status (KRAS). We performed a sub-group analysis by KRAS from the HGCSG0802 observational cohort study that investigated 115 patients (pts) treated with 1st line BV for metastatic colorectal cancer (mCRC). Methods: The objective of HGCSG0802 was to evaluate progression-free survival (PFS), overall survival (OS), response rate (RR), and safety. The key eligibility criteria were with evaluable lesions, older than 20 years, ECOG PS 0-2. In this analysis, pts characteristics, RR and safety were compared using Fisher’s exact test. Univariate and multivariate analysis for PFS and OS were performed using patient characteristics. Survival analyses were performed with Kaplan-Meier method and Cox proportional hazards model. Results: Of 108 pts (the full analysis set), 99 pts were evaluable for KRAS. Sixty-two pts (62.6%) had KRAS wild-type (wt) and 37 pts (37.4%) had mutation (mt). The pts characteristics between those with wt and with mt were generally balanced except for PS 0 (91.9% in wt, 75.7% in mt; p = 0.036) and lung metastasis (33.9% in wt, 62.2% in mt; p = 0.007). RR was 70.0% in wt versus 65.7% in mt. Adverse events related to BV were almost balanced except for bleeding (any grade) (30.6% in wt, 13.5% in mt; p = 0.088). The median PFS and OS was 9.9 and 26.8 months in wt versus 7.9 and 17.5 months in mt (PFS ; HR 1.519, p = 0.064 and OS ; HR 1.944, p = 0.005). In Cox multivariate analysis, KRAS mt showed significantly shorter PFS and OS (PFS ; HR 1.637, p = 0.045 and OS ; HR 2.132, p = 0.005). Conclusions: In this cohort, depending on the KRAS Exon2 mutational status, severe adverse events were no significant difference. The multivariate analysis showed that PFS and OS were significantly longer in the KRAS Exon2 wild-type patients. So, KRAS Exon2 mutational status can be a predictive and prognostic marker in bevacizumab combined 1st line chemotherapy. Clinical trial information: UMIN000018935.
Prospective observational study for the impact of short-term periodic intravenous steroid premedication for gastrointestinal cancer chemotherapy on bone metabolism.
Authors: Ishiguro Atsushi , Nakamura Michio , Muranaka Tetsuhito , Yuki Satoshi , Murai Taichi , Matsuda Chika , Oba Ayane , Itaya Kazufumi , Shigesawa Taku , Koike Yuta , Fujita Yomo , Endo Ayana , Tsukuda Yoko , Ono Yuji , Kudo Takahiko , Nagasaka Atsushi , Nishikawa Shuji , Komatsu Yoshito
Published: February 2017

523

Background: Although intravenous glucocorticoid (GC) premedication (GCP) before chemotherapy (CTx) are frequently used to prevent nausea and vomiting for continuing comfortable CTx, the side effects of intermittent GCs on bone health have not yet been reported. So we designed a multicenter, prospective, observational study to evaluate the impact of periodic GCP on bone metabolism in gastrointestinal cancer (GIC) patients (pts). Methods: The eligibility criteria were as the follows: (i) histologically proven GIC. ; (ii) The duration of periodical GCP is weekly, biweekly, and triweekly. More over 4 weeks GC free intervals is not permissible. ; (iii) age over 20. The primary endpoint was to investigate the variations of bone mineral densities (BMD) at lumbar spine measured by dual energy x-ray absorptiometry (DEXA) and bone turnover biomarkers, cross-linked N-telopeptide of type I collagen (NTX) and bone alkaline phosphatase (BAP), between baseline (BL) and 16 weeks after starting CTx (16w). Results: From June 2013 to April 2015, 98 pts were enrolled. Two pts were not proven as GIC histologically. One patient (pt) was not measured on baseline DEXA. One pt was taken bisphosphonates already on BL point. Four pts were not administered CTx or GCP, and 16 pts were not measured BMD on 16w due to several reasons such as pts refusal, discontinuation of CTx, death and so on (74 pts were full analysis set). In 55 pts (74.3 % of FAS), the levels of BMD at 16w were decreased compared with BL and the average amount of BMD reduction rate was 5.83 % (-38.8 % to 31.1 %). Although no significant difference was found in the level of NTX between BL and 16w (p = 0.118), there was the significant increase of BAP level statistically (p = 0.006). There were also significant correlations between percent change in BMD and NTX, BMD and BAP, NTX and BAP (p = 0.037, 0.029, and 0.003, respectively). Conclusions: We found that periodic GCP in GIC pts caused the reduction of BMD and some influences for bone turnover. These results indicate that GCP might generate more serious osteoporosis of GIC pts during CTx. Further studies are necessary to illustrate the need to prevent GC induced osteoporosis in using GCP. Clinical trial information: 000011054.
RAS testing awareness amongst oncologists and panitumumab prescription for metastatic colorectal cancer: A European physician survey and medical records review.
Authors: Trojan Jorg , Kafatos George , Bennett James , Demonty Gaston , Downey Gerry , Mineur Laurent , Tomasek Jiri , Rouleau Etienne , Fabian Pavel , De Maglio Giovanna , Garcia Alfonso Pilar , Aprile Giuseppe , van Krieken J. Han
Published: February 2017

530

Background: Patients with metastatic colorectal cancer (mCRC) must have a confirmed RAS wild-type status before starting panitumumab (pmab). Two studies, a physician survey and a medical records review (MRR), were carried out to evaluate physician awareness of the need for biomarker testing in mCRC. Methods: Both studies were carried out in three rounds. Rounds 1 and 2 (2012–14) examined KRAS testing only as they were completed before guidelines changed, and have been published previously. Round 3 (2014–15), reported here, was performed after guidelines changed and covered full RAS testing. Participants were from nine European countries:France, Germany, Italy, Spain, the Czech Republic, the Netherlands, Belgium, Denmark, and Sweden. For the survey, oncologists who had prescribed pmab to mCRC patients in the previous 6 months were interviewed by telephone using a standard questionnaire.For the MRR,oncologists were asked to provide clinical information extracted from their patients’ medical records. Results: 152 oncologists and 131 patients’ records were included in Round 3 of the physician survey and the MMR, respectively, compared with 301 oncologists and 306 records in Rounds 1 and 2. In Round 3 of the physician survey, 95.4% (n = 145) of participants correctly reported that pmab should be prescribed in RAS wild-type mCRC only, compared with 99.0% (n = 298) in Rounds 1 and 2, responding to the same question about KRAS testing. In Round 3 of the MRR, 100% (n = 131) of patients had a confirmed RAS or KRAS wild-type status prior to initiation of pmab. Of those patients, 83.2% (n = 109) were tested for RAS status and 16.8% (n = 22) were tested for KRAS status only. Conclusions: Round 3 of the survey demonstrated a high level of knowledge amongst oncologists about the need for RAS testing in mCRC. The MRR supported these findings, showing that this knowledge is being applied to clinical practice. Comparisons between Round 3 and Rounds 1 and 2 show that physician awareness and adherence to prescribing guidelines remained high over time, despite recommendations changing with RAS testing replacing KRAS testing.
Impact of infectious complications on long-term survival following colorectal cancer surgery.
Authors: Mokdad Ali , Hirsch Hannah , Nassour Ibrahim , Borja Nicholas , Balch Glen C. , Polanco Patricio
Published: February 2017

531

Background: Colorectal cancer surgery is associated with significant postoperative morbidity, which may have long-term implications on patient outcomes. We hypothesize that operative complications following surgery for colorectal cancer are associated with increased recurrence and worse survival. Methods: Using a prospectively maintained database, we reviewed patients with colorectal cancer that underwent a curative resection from 2008 to 2015. Patients were categorized by presence of any complication within 90 days from surgery and by type of complication, infectious and non-infectious. We compared clinical, pathological, and perioperative data using t-test, chi-squared test, and ANOVA. We compared overall (OS) and recurrence free survival (RFS) using Kaplan Meier and log-rank test. Multivariable Cox regression was used to compare mortality and recurrence. Results: Two hundred and twenty-nine patients underwent 104 colon and 125 rectal cancer resections (20 pelvic exenterations, 83 low anterior and 17 abdominoperineal resections) were followed for a median of 23 months. Fifty percent were completed minimally invasively. Postoperative complications occurred in 52%; 19% had a major complication (Clavien-Dindo 3-4). Postoperative complications were more likely to occur in open (61% vs. 38%, p < 0.01) and rectal operations (63% vs. 42%, p = 0.02). On multivariable analysis, OS and RFS were not statistically different in patients with complications. Patients with infectious complications had worse 3-year survival when compared to patients with non-infectious complications and without complications (58%,69%,76%, p = 0.04). Recurrence at 3 years was also significantly different among the three groups (p = 0.03). Infectious complications remained associated with worse overall survival (HR 1.8; 95% CI 1.02,3.26) and recurrence free survival (HR 1.9; 95% CI 1.06,3.39) after adjusting for patient, tumor, and perioperative data. Conclusions: Infectious complications following colorectal cancer surgery are associated with worse OS and RFS independent of tumor stage, type of surgery, and technique. Current research is ongoing to explore possible etiologies of this association.
Defective mismatch repair (dMMR) status as a predictive biomarker in Chinese patients (pts) with metastatic colorectal cancer (mCRC) treated with bevacizumab (BV) + chemotherapy.
Authors: Ling Jiayu , Chen Dianke , Zhang Jianwei , Zhong Lixing , Cai Yue , Deng Yanhong
Published: February 2017

537

Background: Monoclonal antibody bevacizumab targeting the vascular endothelial growth factor has become the standard treatment of metastatic colorectal cancer(mCRC) and has led to modest but significant improvements in patient outcomes. Given the modest gains there is a pressing need for predictive factors to better identify patients who would benefit from this target therapy. Methods: We performed retrospective study of 94 Chinese pts with mCRC who received BV combining chemotherapy, whose MMR protein (MLH1, MSH2, MSH6, PMS2) expression was analyzed. Overall survival (OS) was recorded. Univariate and multivariate Cox proportional hazard regression models were used to assess the influence of clinical factors on OS in the cohort. Results: A total of 94 pts (male/female, 58/37; median age, 51.0 years (range 20.0-79.0); synchronous/metachronous metastasis, 70/24; primary lesion resected/unresected 69/25) were enrolled in this study from October 2010 to March 2014. Median OS of the full cohort was 24.5 months (95% CI, 21.2-28.0 months). Of 94 pts, 70 pts were evaluable for MMR. Seven pts (10.0%) had dMMR and 63 pts (90.0%) had pMMR. The pts characteristics between those with dMMR and pMMR were different in age younger than 65 years (100% in dMMR, 69.0% in pMMR; p = 0.08) and synchronous metastasis (100% in dMMR, 75.0% in pMMR; p = 0.13). The median OS was 36.3 months in dMMR versus 21.6 months in pMMR (p = 0.056), and dMMR versus pMMR in bev treating CRC pts was associated with marginal significantly better OS (HR = 0.143; 95%CI, 0.02-1.05; P = 0.056). This association was consistent with that in a multivariable analysis. Also, we found a statistically significant interaction between MMR status and synchronous metastasis (Pinteraction = 0.041). Primary lesion resection was significantly associated with superior OS and was dependent of clinical variables or MMR status. (HRadjusted= 0.43 95% CI 0.20-0.95 P = 0.036 ). Conclusions: MMR status and primary lesion resection status may be predictive factors to better identify patients who would benefit from bevacizumab therapy in Chinese mCRC.
Expression of P4H9-detecting molecule on spindle-shaped fibroblasts to indicate malignant phenotype of colorectal cancer.
Authors: Yokoyama Shozo , Ieda Junji , Yamamoto Naoyuki , Mitani Yasuyuki , Takifuji Katsunari , Hotta Tsukasa , Matsuda Kenji , Watanabe Takashi , Iwamoto Hiromitsu , Deguchi Masaaki , Yamaue Hiroki
Published: February 2017

539

Background: Cancer cells and fibroblasts are coordinated for cancer progression. Our previous study has shown that P4H9, produced by epitope for b2 integrin, detected a molecule on fibroblasts in mammary fat pad mouse model in response to Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) expressing cancer cells. P4H9 detecting molecule (PDM) expression appeared to be associated with myofibroblast differentiation. The aim of the present study is to investigate whether PDM express on fibroblast and cancer cells in clinical tissue samples, and whether PDM expressing cells in colorectal cancer tissue are correlated with clinicopathological features of patients with colorectal cancer. Methods: Immunohistochemistry were conducted with P4H9 on clinical tissue samples from 156 patients with colorectal cancer. The risk factors for metastases and survival were calculated for clinical implication of PDM expressing spindle shaped fibroblasts. Immunofluorescence with P4H9 were performed on CCD-18Co fibroblasts and colorectal cancer cell lines, HT29 and HCT116. Results: Multivariate analysis showed that PDM expressing spindle shaped fibroblasts was an independent risk factor for lymph node metastasis, hematogenous metastasis, and short survival. Kaplan-Meier survival curve presented that PDM expressing spindle shaped fibroblasts was associated with shorter survival time (p< 0.0001). PDM expressing spindle shaped fibroblasts is associated with metastasis and shorter survival of the patients with colorectal cancer. Immunofluorescence showed that PDM expressed on CCD-18Co fibroblasts and colorectal cancer cell lines, HCT116 with fibroblast like morphology. Conclusions: PDM expressing spindle shaped fibroblasts may play a role to bring out malignant phenotype of colorectal cancer. PDM may be a new biomarker and a novel therapeutic target for patients with colorectal cancer.
Esophageal cancer in young patients: Is aggressive treatment associated with better outcomes?
Authors: Glaser Alexander D , Cameron Miles , Almhanna Khaldoun , Hoffe Sarah E. , Fontaine Jacques , Frakes Jessica M. , Pimiento Jose Mario
Published: February 2017

54

Background: Recent studies suggest that survival in young patients with esophageal cancer (EC) is worse than their older counterpart, secondary theoretically to more aggressive disease and late stage at diagnosis. The aim of this study is to compare stage at diagnosis, patterns of treatment and outcomes of young patients with EC at Moffitt Cancer Center (MCC). Methods: Patients with EC treated surgically between 1996 and 2014 at MCC were recorded at an IRB approved database. We divided the patients into two cohorts, those younger or older than 50 years of age. Demographics, pre-treatment clinical data, therapeutic courses and treatment outcomes were analyzed. We compared clinical variables and defined p < 0.05 as statistically significant. Overall survival was examined using the Kaplan-Meier method. Results: Of the 816 patients treated surgically for EC 86 (10.5%) were < 50 years old. The majority of patients were male 622(85.2%) vs. 67(77.9%) in the younger group (p =.084). Of the 816 patients, 569 had pre-treatment staging information available. Pre-treatment clinical staging for the older and younger cohorts was, Stage0/TIS = 13(2.2%) vs. 1(1.4%); Stage I = 90(15.3%) vs. 6(8.3%); Stage IIA = 119(20.3%) vs. 16(22.2%); Stage IIB = 88(15.0%) vs. 8(11.1%); Stage III = 269(46%) vs. 40(55%); Stage IV = 8(1.4%) vs. 1(1.3%)(p =.44). Younger patients were more likely to receive neoadjuvant therapy (75.6% vs.62.1%)(p =.013), and also neoadjuvant combined with adjuvant therapy, 24(41.4%) compared to 48(12.1%) in the older cohort (p =.001). Pathological complete response (pCR) rate was higher in older patients (43.2% vs 32%)(p =.204); Median survival was higher in younger cohort 64.3 vs. 56.4 months. Neither of these were statically significant (p =.516). Conclusions: This retrospective study with several variables only included surgical patients. In our experience, younger age was not associated with late stage presentation. Younger patients were more likely to be treated with neoadjuvant therapy, and despite similar pathologic response rates they were more likely to receive adjuvant therapy. Although younger patients received more aggressive treatment, their overall survival did not differ from their older counterparts.
RAS mutational status and CEA production at initial presentation in metastatic colorectal cancer.
Authors: Cho May Thet , Goldstein Leanne , Akiba Chie , Lau S. Cecilia , Telatar Milhan , Afkhami Michelle , Sentovich Stephen , Melstrom Kurt , Lim Dean , Chao Joseph , Chung Vincent M. , Fakih Marwan
Published: February 2017

542

Background: Serial CEA testing is recommended in the surveillance of patients with resected stage II-IV colorectal cancer. However, the sensitivity of CEA in identifying metastatic disease has not been evaluated in the settings of RAS mutant (MT) and RAS/BRAF wild-type tumors (WT). In order to evaluate the impact of RAS mutational status on CEA production, we retrospectively evaluated a single-institute metastatic colorectal cancer (mCRC) population. Methods: We retrospectively reviewed, in a single center, all cases with mCRC with known RAS mutational status based on next generation sequencing (ONCO44 and ONCO48). These assays identify clinically relevant mutations in BRAF, KRAS, and NRAS. Additional eligibility criteria included the availability of CEA levels and imaging studies at first diagnosis of mCRC. Patient demographics, primary tumor location and sites of metastatic disease at 1st diagnosis were captured. CEA levels were stratified as normal or elevated based on a cut point of 5ng/ml. Results: 139 mCRC patients satisfied the eligibility criteria (75 RAS-MT, 59 RAS/BRAF-WT, and 5 BRAF-MT). BRAF-MT patients were excluded from the analysis due to their small sample size. Patients with RAS/BRAF-WT tumors were more likely to present with metastatic disease to the liver, but this did not reach statistical significance (p = 0.056). There was no difference in the incidence of normal CEA at presentation in RAS-MT (30%) and RAS/BRAF-WT (28%) cohorts. CEA production was dependent on the pattern of metastatic disease. Elevated CEA was associated with the presence of liver metastases versus no metastases among RAS-MT (92% vs 47% p <.0001) and RAS/BRAF-WT patients (82% vs 50% p = 0.0101). RAS status did not impact the likelihood of CEA production within the hepatic metastases and non-hepatic metastases groups. Conclusions: RAS status does not appear to influence CEA production in patients with mCRC. CEA elevations are highly associated with liver metastases and are less prevalent in patients without hepatic involvement. These findings confirm the limited predictive value of CEA for non-hepatic recurrence, irrespective of RAS status.
Early-onset small bowel and colorectal cancer patients with BRCA mutations: Actionable variants in atypical presentations.
Authors: Vikstrom Karen
Published: February 2017

545

Background: The paradigm of genetic panel testing for hereditary colorectal cancer (CRC) continues to emerge. An association between Hereditary Breast and Ovarian Cancer (HBOC) syndrome and early onset CRC and has been reported, though only significantly associated with BRCA1 mutations. Guidelines do not include CRC in the HBOC spectrum. We describe nine early-onset ( < 55 years) gastrointestinal (GI) cancer patients with germline mutations in BRCA1 or BRCA2 to highlight actionable findings that would have been missed by traditional CRC genetic testing. Methods: The nine affected patients underwent genetic testing using NGS-based multi-gene cancer panels. Genomic DNA variants were identified and classified according to ACMG criteria. Patient medical histories were obtained by referring clinician and documented on test requisition forms, and were de-identified for analysis. Results: Nine patients had a pathogenic (P) variant in BRCA1 (three cases) or BRCA2 (six cases). Four patients were under the age of 32 at the time of their CRC diagnosis. Of the nine cases, only one met NCCN testing criteria for HBOC syndrome. All patients were negative for P variants in the canonical CRC genes (APC, MUTYH, MLH1, MSH2, MSH6, PMS2, EPCAM). Conclusions: In this series, nine unrelated patients with early-onset CRC were found to carry a BRCA1 or BRCA2 mutation. The presence of BRCA mutations in these CRC patients is alone not strong evidence of causation; however, it suggests that clinicians should consider expanded panel testing in the presence of complex family histories, as these patients would have been found negative by traditional genetic tests. As the phenotypic spectrum of classical cancer syndromes is expanded by broad panel-based testing, it is important to identify patients that fall outside traditional diagnostic criteria, as some patients will have actionable findings in unexpected genes.
The prevalence of colorectal polyps in patients with gastric cancer.
Authors: Kwon Kyoung-Joo , Shim Ki-Nam , Jung Sung-Ae , Jung Hye-Kyung , Kim Seong-Eun , Kong Kyoung Ae
Published: February 2017

549

Background: Gastric polyps are found frequently in various colonic polyposis syndromes. Genetic alterations of several genes occur in gastric adenomas and colorectal adenomas. However, it is unknown whether patients with gastric cancers are at higher risk for colorectal adenomas. Methods: We retrospectively reviewed the medical records in 112 patients with gastric cancers from January 2005 to December 2009. Patients who were diagnosed with gastric cancers and had colonoscopy within 6 months were compared to 220 sex and age-matched controls from 3,725 persons who underwent esophagogastroduodenoscopy and colonoscopy for screening at the Center for Health Promotion in Ewha Womans University Medical Center. High risk polyp was defined by any of the following: (1) the presence of three or more polyps; (2) polyp size at least 10mm; (3) high-grade dysplasia or adenocarcinoma confirmed by histopathologic examination. Otherwise the polyps were defined as low risk. Results: Mean age was 59 years-old and 70% was male in patients with gastric cancer. Overweight (BMI ≥ 23) was more common in control group (gastric cancer versus control: 56% and 69%, respectively, P = 0.025). Colorectal polyps were identified in 67 of 112 patients (59.8%) with gastric cancer and in 153 of 220 controls (69.5%). The prevalence of high risk polyps were numerically higher in the gastric cancer group than the control group, although not statistically significant (9% vs. 4%, P = 0.267). In contrast, the prevalence of low risk polyps were significantly frequent in the control group (91% vs. 96%, P = 0.024). The mean size and number of colorectal polyps were not different between the two groups. Adenocarcinoma was diagnosed from polyps in five patients (7%) with gastric cancer and four patients (2%) with control group (P = 0.190). Conclusions: The overall colorectal polyps, especially low risk polyps, were more prevalent in the control group. In contrast, high risk polyps and adenocarcinoma is more frequently found in patients with gastric cancer although not significant. Therefore, we suggest that colorectal polyps should be cautiously examined in patients with gastric cancer.
Validation of the prognostic impact of lymphocyte infiltration (LI) in patients (pts) with stage III colon cancer (CC) treated with adjuvant FOLFOX+/- cetuximab: A PETACC8 translational study.
Authors: Emile Jean-François , Le Malicot Karine , Tabernero Josep , Mini Enrico , Folprecht Gunnar , Van Laethem Jean-Luc , Julie Catherine , Allard Marc Antoine , Bennouna Jaafar , Lepage Come , Laurent-Puig Pierre , Taïeb Julien
Published: February 2017

553

Background: The prognostic value of LI of CC has been demonstrated by several groups. However no validated test is available for clinical practice. We previously described an automated and reproducible method for testing LI (Allard MA et al 2012) and aimed to validate it for clinical use. Methods: According to NIH criteria, we designed a prospective analysis of this biomarker in pts included in the PETACC8 phase III study. Primary objective was to compare % of pts without recurrence at 2 years in pts with high versus low LI (#NCT02364024). Secondary objectives were comparison of disease free (DFS) and overall (OS) survivals, and prognostic value of LI on these endpoints. Automated testing of LI was performed on virtual slides without access to clinical data. Results: Among the 1,220 CC pts enrolled, LI was high, low and not evaluable in 241 (20%), 790 (65%) and 189 (15%), respectively. High and low LI groups did not differ except for treatment arm (p=0.04) and microsatellite status (MSI/MSS) status (p=0.04). Primary objective was met with a 2-year recurrence rate of 14.4% versus 21.1% in pts with high and low LI respectively (p=0.02). Pts with high LI also had better DFS and OS (table). Stage, grade, KRAS status and LI were the only prognostic markers in multivariate analysis. Sub-group analyses revealed that high LI had better DFS and OS in MSS pts, and in pts without KRAS codon 12-13 mutation, but not in MSI neither in KRAS mutated pts. Conclusions: This is the first prospective validation of a LI testing in pts with stage III CC treated with FOLFOX adjuvant. Automated testing of LI also had a prognostic value for DFS and OS. Funding: Merck-Serono, Sanofi, Région Ile de France. Clinical trial information: NCT02364024. [Table: see text]
Genomic features of primary and recurrent anal squamous cell carcinoma.
Authors: Pike Jonathan W , Mouw Kent William , Braunstein Lior Zvi , Martin Neil E. , Mamon Harvey J. , Hornick Jason L. , Van Allen Eliezer Mendel , D'Andrea Alan D.
Published: February 2017

556

Background: Approximately 7000 cases of anal squamous cell carcinoma (ASCC) are diagnosed annually in the US, and the incidence is increasing. Combination chemoradiotherapy (CRT) is the standard of care for locally-advanced ASCC, and the 5-year survival rate is approximately 70%. Unlike many other tumor types, no large-scale genomic studies have been reported for anal cancer. In order to characterize the mutational landscape of ASCC and identify potential therapeutic targets, we perform comprehensive genomic analysis of a pilot cohort of ASCC cases. Methods: We performed whole exome sequencing of tumor and matched germline DNA from a pilot cohort of twelve patients with locally advanced (Stage II-IVA) ASCC cases treated at Dana-Farber Cancer Institute. All patients received concurrent chemoradiotherapy (CRT): seven ‘responders’ had complete response and no evidence of recurrence with a minimum of one year follow-up, while five ‘non-responders’ had residual or recurrent disease following CRT. For non-responders, both primary and residual/recurrent tumors were analyzed. Results: The overall mutation rate was 3.7 mutations per megabase (Mb). Recurrent mutations in several known cancer genes were observed. Five of twelve cases had a hotspot mutation in FBXW7, including 3 of the 5 non-responders. Known oncogenic mutations were also observed in PIK3CA (3 tumors) and NFE2L2/KEAP1 (3 tumors). Analysis of paired primary and recurrent samples revealed surprising examples of distinct driver mutations in clonally-related tumors. Copy number analysis revealed focal amplifications of chromosome 3q. Conclusions: This study represents one of the first genome-scale analyses in ASCC. The overall mutation rate was similar to other HPV-associated squamous cell carcinomas, and recurrent mutations in several known cancer genes were observed. Analysis of paired primary and residual/recurrent tumors revealed surprising heterogeneity. To extend our findings, we are currently performing a similar analysis in a larger extension cohort of ASCC tumors.
Novel driver genes and genomic predictors of immune infiltrates in colorectal cancer.
Authors: Giannakis Marios , Mu Jasmine Xinmeng , Shukla Sachet , Qian Zhi Rong , Cohen Ofir , Nishihara Reiko , Cao Yin , Yamauchi Mai , Sukawa Yasutaka , Nosho Katsuhiko , Ng Kimmie , Meyerhardt Jeffrey A. , Lawrence Michael , Gabriel Stacey B. , Lander Eric , Getz Gad , Wu Catherine , Fuchs Charles S. , Ogino Shuji , Garraway Levi A.
Published: February 2017

557

Background: Despite progress that has been made in the molecular characterization of colorectal cancer (CRC), prior studies have had limited power to detect, as significantly mutated, less-frequently altered CRC genes. In addition, most large-scale sequencing studies have generally lacked clinicopathologic annotations to link genomic features with important CRC characteristics such as immune infiltration. Methods: We performed Whole-Exome Sequencing on 619 archival primary CRCs from two large prospective cohorts, the Nurses’ Health Study and the Health Professionals Follow-Up Study. We identified CRC driver genes using significance-calling algorithms and employed a novel computational pipeline to calculate tumor neoantigens (peptides resulting from somatic mutations and recognized by the immune system as foreign). The neoantigen load and altered genes/pathways were correlated to immunohistochemically determined immune infiltrates and survival data linked to these specimens. Results: We identified 90 significantly mutated genes in non-hypermutated CRC. These include previously underappreciated genes that participate in pathways such as WNT-signaling and RNA processing. Among all samples, the neoantigen load was associated with increased overall lymphocytic infiltration (P = 2.6e-11), tumor-infiltrating lymphocytes (TILs) (P = 2.0e-19), memory T-cells (P = 0.091) and CRC-specific survival (P = 0.025). Neoantigen load was also associated with higher TILs in microsatellite-stable tumors (P = 0.015). We found that HLA and Antigen Processing Machinery (APM) mutations have undergone positive selection in tumors with TILs. Conclusions: In the largest exome-sequencing study of CRC to date, we leveraged the increased number of samples and their associated clinicopathologic annotations to identify new driver genes and genomic predictors of immune infiltrates in this disease. Neoantigen load is prognostic in CRC and predictive of immune infiltration, even among microsatellite-stable tumors. We also find evidence of positive selection for HLA and APM mutations in immune-cell infiltrated tumors. These results have implications for patient selection in CRC immunotherapy trials.
Effect of age and ethnicity on the tumor location, nuclear accumulation of p53 and clinical outcome in colorectal adenocarcinoma.
Authors: Paluri Ravi Kumar , Behring Michael , Posey James , Manne Upender
Published: February 2017

559

Background: The race/ethnicity based research in colorectal cancer (CRC) care continues to remain a high priority in developing personalized medicine, and to improve overall clinical outcomes. The role of p53 abnormalities in prognostication of CRC has been evaluated earlier. The incidence of nuclear accumulation of p53 (p53nac) and its prognostic relevance in African American (AA) and non-Hispanic white patients (pts) have been investigated, and it was suggested that the clinical consequences of p53nac in CRC varies with anatomic location of the tumor and the race of the patient. However, the clinical value of p53nac in relation to age, the tumor location, and race together is not assessed. Thus, we evaluated prognostic significance of p53nac by considering the tumor location, age, race/ethnicity and p53nacin CRCs in AA and white pts. Methods: Formalin fixed paraffin embedded CRC tissues from 242 AAs and 346 whites who underwent surgery were assessed for p53nac by routine immunohistochemistry (IHC). The routine (not antigen retrieval) IHC will identify the majority of genetic alterations ( > 95% missense point mutations) and have significant association with patient survival in CRC. The association between phenotypes, p53nac status, clinicopathologic features, and overall survival were evaluated using the x2 test and Cox regression analyses. Results: Approximately equivalent proportions of distal (52%) and proximal adenocarcinomas (48%) were positive for p53nac in AA pts. In contrast, distal CRC from whites more frequently were positive for p53nac than from the proximal colon (67% vs. 34%, x2 P = 0.006). p53nac was found to be a strong predictor of poor overall survival in young ( < 65 yr) white pts with proximal tumors [hazard ratio (HR) = 2.8, 95% Confidence Intervals (CI):1.2-6.4] but not in AAs (HR = 0.7, 95% CI: 0.41-1.21). Conclusions: The findings of this study suggest that p53nac is a strong prognostic marker for young white pts with proximal colon adenocarcinomas. Our findings are clinically relevant because several small-molecule inhibitors of mutant p53 are under investigation. These studies were supported by a pilot project grant by the UAB Comprehensive Cancer center.
Effect of combined consideration of distinct signatures of VEGF and soluble VEGFR2 on prognostic implication in gastric cancer.
Authors: Ock Chan-Young , Nam Ah-Rong , Bang Ju-Hee , Kim Tae-Yong , Lee Kyung-Hun , Han Sae-Won , Im Seock-Ah , Kim Tae-You , Bang Yung-Jue , Oh Do-Youn
Published: February 2017

56

Background: Anti-angiogenic strategy in gastric cancer (GC) has been highlighted again due to the recent success of ramucirumab and apatinib. Therefore, the comprehensive network of VEGF, soluble VEGF receptor-2 (sVEGFR2) and cytokines and other angiogenic factors (CAF) in GC and their prognostic impact would be of importance, although they have been poorly understood. We aimed to find out the CAF signature associated with VEGF and sVEGFR2, and to explore their prognostic implication in GC. Methods: We measured pretreatment serum levels of 52 CAFs, including VEGF and sVEGFR2, using multiplex bead immunoassays and ELISA, in 70 patients who were diagnosed with GC in Seoul National University Hospital, and treated with palliative chemotherapy. Linear regression analysis for correlating CAFs with VEGF and sVEGFR2, and survival analysis by log rank test and Cox regression analysis were performed. Results: The VEGF signature was shown to be associated with seven CAFs (interluekin [IL]-7, IL-12p70, IL-2Ra, IL-10, stem cell factor, Fibroblast growth factor-basic, IL-3). The sVEGFR2 signature was associated with IL-4 and platelet-derived growth factor beta, but VEGF and sVEGFR2 showed no association with each other. Patients with high VEGF had a tendency to have worse overall survival (OS) than those with low VEGF (11.2 months versus 16.7 months; P = 0.061). However, among patients with high-sVEGFR2, OS was not different according to VEGF (12.1 months, high-VEGF versus 15.1 months, low-VEGF; P = 0.546). Interestingly, the poor prognostic impact of high-VEGF was far significant in patients with low-sVEGFR2 (10.9 months versus 16.8 months; P = 0.036). With this perspective, VEGF/sVEGFR2 ratio was significantly correlated with worse OS in univariate as well as multivariate analysis (HR 1.78 [95% CI 1.08-2.94], P= 0.024). Conclusions: Based on the comprehensive network analysis of CAF, VEGF and sVEGFR2 had distinct CAF signatures in GC. Consideration of both VEGF and sVEGFR2 confers more accurate prognostic implication compared with VEGF alone in GC. Regarding the angiogenic aspect, VEGF/sVEGFR2 ratio is significantly correlated with survival outcome in GC.
Clusterin expression (CLU) as a prognostic marker in colorectal carcinoma (CCR).
Authors: Alcaide Julia , Rueda Antonio , Rodrigo Isabel , Tellez Teresa , Funez Rafael , Padilla Maria del Carmen , Zarcos Irene , Pérez-Ruiz Elisabeth , Villatoro Rosa , Perez Diego , Redondo Maximino
Published: February 2017

563

Background: Increased CLU is involved in malignant progression and anticlusterin treatment with antisense oligonucleotides enhances apoptosis induced by several citotoxics. However, clinical significance of CLU expression in human CRCs has been scarcely studied. We investigated whether changes in CLU could be related to carcinogenesis and survival (sv) of CRC patients (pts). Methods: Formalin-fixed and paraffin-embedded specimens were examined from 31 adenomas and 103 CRCs resected at Costa del Sol Hospital. The study was approved by Research Ethics Committee. Immunohistochemistry using monoclonal anti-α chain clusterin antibody (Upstate-Millipore, Watford, England) was performed, following standard staining procedure. CLU was scored as negative (CLU–) (no staining) or positive (CLU +) (>10% of tumor cells with strong staining). Cytoplasmic CLU in tumors was evaluated for cancer cells only, and in normal mucosa for epithelial cells only. Sv curves were calculated and plotted according to Kaplan-Meier method. Predictors that were significant at p<0.10 in univariate analysis, were entered into a Cox proportional hazards model for multivariate analysis, remaining significant at p<0.05. Results: Median follow-up was 54 months. Median age was 70 years (45-91). TNM stage distribution was: I (13%), II (48%), III (25%) and IV (14%). Epithelial normal cells were always CLU-, but 16% (5/31) of adenomas was CLU+ and this percentage increased in CRCs (30%, 31/103). Positive staining always presented an apical cytoplasmic pattern. Recurrence was more frequent in CLU+ (61%,19/ 31) than in CLU- tumors (37%, 27/72) and CLU was significantly associated with lower disease-free survival (DFS) (p<0.05). In multivariate analysis, CLU and stage remained significant independent prognostic factors for DFS (Table). Conclusions: CLU has a role in colon carcinogenesis and prognostic value. CLU is associated with decreased DFS among pts with CRCs. These findings have important implications for identifying CRC pts with more aggressive tumors who may benefit from targeted therapy against clusterin. [Table: see text]
Comprehensive genomic profiling in colorectal cancer (CRC) to identify differing frequencies of clinically relevant genomic alterations (CRGA) in tumors of patients (pts) less than age 50 as compared to those of pts over age 65.
Authors: Bailey Mark , Mitchell Edith P. , Morosini Deborah , Lipson Doron , Ross Jeffrey S. , Miller Vincent A. , He Jie , Stephens Phil
Published: February 2017

570

Background: CRC is increasing in young pts and are believed to have worse pathological features than older pts. Mutational profiles may differ in pts. The objective of this study was to compare and describe the genomic complexity of CRC in pts < 50 to > 65 in the FoundationOne database. Methods: DNA was extracted from 2,267 FFPE clinical specimens with colorectal cancer (CRC) (< 50 yo, n=1280; > 65 yo, n = 987). CGP was performed on hybridization-captured, adaptor ligation based libraries to a mean coverage depth of 603X for 3,769 exons of 236 cancer-related genes plus 47 introns from 19 genes frequently rearranged in cancer. All classes of genomic alterations (GA) were identified. Clinically relevant genomic alterations (CRGA) were defined as GA linked to drugs on the market or under evaluation in mechanism driven clinical trials. Results: Among the 50 genes analyzed, the > differences were noted in BRAF and FAM123B in pts > 65. KRAS mutations occurred in 50.5% vs 51.3%; NRAS, 2.6% vs 4.8%, BRAF, 5.5% vs 11.9% in younger pts vs older pts, respectively. Few differences in genomic complexity were observed, although a trend toward increase frequency of BRAF mutations were observed in pts > 65. Conclusions: While some differences in tumor genomic profiles and complexity were observed in CRC pts < 50 vs > 65, changes in BRAF and FAM123B were more frequent in the > 65. Ongoing studies will assess differences in molecular and genomic features between younger and older CRC pts. [Table: see text]
Possible role of the systemic inflammatory reaction in defining tumor responder vs. nonresponder in cancer macrobead therapy.
Authors: Smith Barry H. , Andrada Zoe Padua , Nazarian Angelica , Ocean Allyson J. , Parikh Tapan , Lu Jia , Arreglado Anna , Fahey Thomas J. , Berman Nathaniel , Gazda Larry , Martis Prithy , Laramore Melissa , Akahoho Eugene , Thomas Joanne , Wolf David J.
Published: February 2017

572

Background: Peritoneal implantation of mouse renal adenocarcinoma cell-containing (RENCA) Macrobead (MB) represents a cell-system-based approach to the treatment of advanced, mCRC that has been evaluated to date in Phase IIa trials. The data indicate that there are “responders” (R) and “non-responders”(NR) as reflected in overall survival (OS), where “response” is defined as a >20% decrease in either/both CEA or CA19-9 during the first 30 days after MB implantation. We analyzed whether the “response” is due to a post-implant systemic inflammatory response (SIR) or rather a direct inhibitory effect of the MB. Methods: Thirty-four treatment-resistant mCRC patients (pts) were implanted laparoscopically at least once with RENCA MB. Pts were considered R (n=25), or NR (n=9), based on tumor marker responses within the first 30 days. CRP, IL-6, TNF-alpha, and ESR, as measures of SIR, were measured at Day 14 and 30. Results: All 34 pts showed SIR to MB implantation, as indicated by transient rises in CRP, IL-6, TNF-a, and ESR. Baseline CRP values (R, mean 3.24+/-4.39 vs. NR, 2.96+/-3.43; t-test, p=0.86), Day 14 CRP values (R, mean 20.97 +/- 7.21 vs. NR, 14.5+/-8.78; t-test, p=0.04), Day 30 CRP values (R, mean 8.21+/-5.43 vs. NR, 10.76+/-6.92; t-test, p=0.27) and mean changes in IL-6 (baseline p=0.28; Day 14 p=0.36; Day 30 p=0.54), TNF-a (baseline p=0.37; Day 14 p=0.32; Day 30 p=0.29) did not show statistically significant differences between R and NR groups. Conclusions: Data suggest that early tumor marker decreases in R of RENCA MB are likely not due to the induced SIR, but rather a possibly direct anti-tumor-cell effect by factors released by MB. This supports the importance of the MB-induced changes in the MEF-2 pathway in the target colorectal cancer cell/tumor reported previously. Studies of clinical efficacy of MB continue in a Phase IIb clinical trial. Clinical trial information: NCT01053013.
The prognostic role of the host immune response in rectal cancer patients treated with neoadjuvant chemoradiation.
Authors: Sprenger Thilo , Wirtz Ralph M , Pektas Selcan , Bohnenberger Hanibal , Kauffels Anne , Conradi Lena-Christin , Homayounfar Kia , Liersch Torsten , Ghadimi Michael
Published: February 2017

575

Background: The development of metachronous distant metastasis is – despite remarkable progress - still a challenging problem after multimodality treatment of rectal cancer. The host immune response has been identified as a valid prognostic marker in primary colon cancer but not in rectal cancer after neoadjuvant chemoradiotherpy so far. Methods: We analysed the mRNA and protein expression of the B- and T-cell chemokines CXCL13 and CXCL9 as well as the B-cell marker IGHM in 77 rectal cancer specimens undergoing preoperative chemoradiotherapy and investigated the role of the host immune response on the occurrence of distant metastasis and survival. Results: The intratumoral mRNA expression levels of the T-cell activator CXCL9 (p < 0.001) as well as the B-cell chemokine CXCL13 (p < 0.01) were inversely correlated with the development of distant metastasis. Patients with high CXCL9 (p < 0.0001) or CXCL13 expression (p < 0.01) had a significantly prolonged DFS. Moreover, patients with high mRNA levels of both CXCL9 and CXCL13 had an excellent DFS compared to patients without expression of any of the chemokines (p < 0.0001). Additionally, CXCL9 expression was correlated with an improved CSS (p < 0.05). There was no significant correlation between the B-cell marker IGHM and metastasis or survival. Conclusions: The host immune response represented by intratumoral expression levels of the lymphocyte-activating chemokines CXCL9 and 13 is associated with reduced rates of distant metastases and improved survival in rectal cancers after multimodality treatment. Further investigations are needed to prospectively validate these results and characterize the underlying cellular and molecular mechanisms.
Novel probe-based quantative image of mitochondria in live colon cancer tissues by multiphoton microscopy.
Authors: Kim Eun Sun , Chun Hoon Jai , Jeen Yoon Tae , Keum Bora , Choi Hyuk Soon , Kim Seung Han , Lee Jae Min , Yoo In Kyung
Published: February 2017

581

Background: Multiphoton endomicroscopy is the recentely updated technique for endoscopy and virtual image and optical sectioning. However optimized probe has not been established for multiphoton endomicroscopic image. Therefore we developed novel probe for mitochondria and applied for colon neoplasm tissues. In cancer cell, abnormally increased mitochondrial replication is related mitochondrial dysfunction and Warburg effect. Methods: We used newly developed multiphoton probe for micochondria imaging which are made using benzofuran derivative (BFP, maximal multiphoton fluorescence at 570 nm). Fresh mucosal tissues of colonic adenoma and adenocarcinoma were obtained from endoscopic biopsy. Multiphoton probe BFP for mitochondria was stained for tissues and imaging performed using multiphoton microscopy. Results: BFP shows high enhancement factor upon binding mitochondria, good selectivity, cell permeability, and can readily detect mitochondria in human tissues by multiphoton microscopy. Mitochondria were detected in human colon mucosa tissues. Calculated mitochondria area were increased in adenocarcinoma tissues compared to normal mucosal tissues. Conclusions: Newly developed multiphoton probe for mitochondria are usable to image human live colon tissues.