bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-6/STAT3 signaling impaired induction of cancer-antigen specific T cells via down-regulation of dendritic cells in tumor microenvironment.
Authors: Ohno Yosuke , Kitamura Hidemitsu , Ohtake Junya , Kaneumi Shun , Sumida Kentaro , Homma Shigenori , Kawamura Hideki , Takahashi Norihiko , Taketomi Akinobu 
Published: February 2017

590 Background: Immunosuppression in tumor microenvironments is a critical issue for cancer immunotherapy. Recently, the effectiveness of immuno-check point therapy has been reported on various types of solid tumors. But, the effectiveness in colorectal cancer was poor compared to other cancers, such as melanoma and renal cell carcinoma. Correct regulation of dendritic cell (DC) function in tumors is important for inducing anti-tumor immunity. We have been demonstrated that IL-6 inhibits antigen presentation by DCs through STAT3 activation in tumor-bearing mice. In this study, we focused on the role of the IL-6/STAT3 signaling cascade in human DCs. Methods: Both IL-6 and pSTAT3 expressions in tumor sites of colorectal cancers was verified by immunohistostaining. CD11b+CD11c+DCs in cancer tissues and PBMCs were isolated by fluorescence-activated cell sorting system and investigated about the surface molecules such as HLA-DR, T cell stimulating ability, and effector gene expression levels. Moreover, we investigated influence of IL-6/STAT3 signaling in human DCs in vitro. Results: The results of IHC revealed that IL-6 was preferentially produced by cancer-associated fibroblasts and immune cells in the tissues of colorectal cancers. In addition, it was confirmed that STAT3 was activated in tumor-infiltrating immune cells. Tumor infiltrating CD11b+CD11c+ DCs highly induced IL-6 gene, down-regulated surface expression of HLA-DR, and attenuated T cell stimulating ability. In vitro experiments showed that IL-6-mediated STAT3 activation reduced surface expression of HLA-DR. COX2, cathepsin L (CTSL), and arginase activity were are involved in the IL-6-mediated down-regulation of surface expression levels of HLA-DR expression levels on DCs. Gene expressions of CTSL, ARG1 as well as IL6 in tumor infiltrating CD11b+CD11c+DCs were much higher than those of PBMCs. Conclusions: IL-6-mediated STAT3 activation inhibits functional maturation of DCs, causing suppression of anti-tumor immunity in colorectal cancer. Therefore, inhibition of the IL-6/STAT3 signaling pathway could be a promising strategy for improving immunotherapies for colorectal cancer patients.
Evaluation of the CpG island methylator phenotype as a predictive biomarker for adjuvant oxaliplatin-based chemotherapy in colorectal cancer.
Authors: Shiovitz Stacey , Wu Chen , Yu Ming , Gourgioti Georgia , Raptou Georgia , Kotoula Vassiliki , Pentheroudakis George E. , Kalogeras Konstantine T. , Fountzilas George , Grady William M. 
Published: February 2017

592 Background: The CpG island methylator phenotype (CIMP) has previously been shown to be a predictive biomarker for 5-fluorouracil (5-FU)/leucovorin (LV)/irinotecan (IFL) adjuvant chemotherapy in stage III colon cancer patients. It is unknown if CIMP serves as a biomarker for standard-of-care oxaliplatin-based adjuvant therapy. Methods: HE6C05 randomized 441 patients with stage II/III colorectal adenocarcinoma to adjuvant oxaliplatin with either capecitabine (XELOX) or 5-FU/LV (mFOLFOX6) between November 2005 and January 2008. The primary objective was disease-free survival (DFS); overall survival (OS) was a secondary objective. The study closed early due to poor accrual with last follow-up in November 2013. Isolated DNA from 293 tumor samples was used to determine CIMP status based on methylation patterns at the CACNA1G, IGF2, NEUROG1, RUNX3, and SOCS1 loci. Cox univariate and multivariate models were used to assess the effects of CIMP on DFS and OS, accounting for interactions with TNM stage; site (right vs. left); KRAS, BRAF, and mismatch repair (MMR) status; tumor T-cell ratio (CD3/CD8); and treatment arm. Results: Of the tumor samples available for CIMP analysis, 28 (9.6%) were CIMP-positive. The CIMP-positive tumors were more likely to be right-sided and have BRAF mutations (chi-square, p<0.001). In the univariate analysis, no significant differences in DFS or OS were observed between individuals with CIMP-positive vs. negative tumors (OS hazard ratio 1.27, Wald’s p=0.55). In addition, CIMP had no predictive value for response to the treatments administered in the two arms of the trial (XELOX vs. mFOLFOX6). In the multivariate model, only TNM stage and primary site were associated with survival outcome (Wald’s p=0.0002 and p=0.006, respectively); CIMP did not improve treatment response prediction. Conclusions: In this exploratory analysis, unlike what we have seen with IFL, CIMP does not appear to serve as a predictive biomarker for treatment response to oxaliplatin-based therapy. CIMP analysis in an irinotecan vs. oxaliplatin trial may offer additional insight.
Prognostic value of RAS, BRAF, and PIK3CA mutations in early-stage colorectal cancer.
Authors: Byer Jennifer Elizabeth , Kothari Nishi , Juan TzuHua , Teer Jamie K. , Schell Michael J. , Shibata David , Kim Richard D. 
Published: February 2017

594 Background: Activating mutations in the KRAS, BRAF and PIK3CA oncogenes deregulate growth-factor pathways and promote metastasis in colorectal cancer (CRC). The prognostic value of these mutations has been reported with conflicting results in early CRC. We evaluated RAS, BRAF, and PIK3CA mutations as prognostic biomarkers in early stage (stage I-III) colorectal cancer (CRC) patients. Methods: Tumor samples collected from 302 early stage CRC patients diagnosed between 1998 and 2010 were analyzed as part of a multi-institutional observational study. Targeted exome sequencing was performed using the Illumina NGS platform with 50-100X coverage of mutations. The BWA/GATK pipeline was used to identify variants and indels. Matched normal samples were not available for comparison to identify somatic mutations, therefore 1000 Genomes was used to filter normal variants. Variants identified in 1000 Genomes with an MAF <0.01 were filtered. Overall survival data was collected via retrospective chart review. Extended RAS, BRAF V600E, and PIK3CA (exon 9 and 20) mutations were evaluated. The log-rank test was used to compare survival distributions. Results: 302 patients were eligible for analysis (53 stage I, 125 stage II, 124 stage III). 109 patients had RAS mutations (KRAS or NRAS), 41 patients had BRAF mutations, and 29 patients had PIK3CA mutations. Of the 247 patients with microsatellite stability (MSS) 98 were RAS mutant, 10 were BRAF mutant, and 19 were PIK3CA mutant. Of the 55 patients with microsatellite instability high (MSI-H) 11 were RAS mutant, 31 were BRAF mutant, and 10 were PIK3CA mutant. BRAF mutation was prognostic for decreased OS (p= 0.0245), particularly in patients with MSS tumors (p=0.0141). RAS and PIK3CA mutations did not have prognostic value for OS (p =0.72 and 0.23 respectively). Conclusions: In early stage colorectal cancer, we confirmed BRAF mutation is prognostic for OS particularly in MSS patients. RAS and PIK3CA mutations did not confer prognostic value.
Detection of tumor progression via cell-free DNA (cfDNA) in patients with colorectal cancer.
Authors: Mende Marika , Thiede Christian , Schuster Caroline , Aust Daniela Ellen , Folprecht Gunnar 
Published: February 2017

598 Background: Individualized therapy based on molecular stratification has become an integrative part in modern targeted treatment approaches. Here, we aimed to assess whether repeated measurements of cfDNA in patients' plasma could indicate tumor progression. Methods: We prospectively collected and stored serum from patients with colorectal cancer presenting for treatment or follow-up. KRAS and NRAS mutations were determined in the formalin fixed, paraffin embedded initial tumor material. A group of patients with confirmed KRAS or NRAS mutation, radiologically proven tumor progression and subsequent change of therapy was selected. cfDNA was extracted from samples taken at time of progression, as well as day -30 and day -60. Mutations were analyzed using a highly sensitive method based on ultradeep Next Generation Sequencing on a semiconductor-based NGS system (IonTorrent PGM), with a lower limit of detection down to 0.01% for selected mutations. Results: Twenty patients were identified to meet the inclusion criteria; 17 were KRAS mutated, three were NRAS mutated within the initial tumor material. In three patients (15%), RAS-mutation was not detectable at time of progression. While in two of them RAS-mutation had been measurable in prior samples, in one case the mutation was not detectable in cfDNA at any time. In 11 (65%) of the remaining 17 patients, RAS variant allele frequency (VAF) in the cfDNA was increased at time of progression compared to measurements at days -30 and/or -60. The median slope of increase was 3.5 (range 0.12 to 15.13). In six patients, there was no correlation between tumor progression and cfDNA; The amount of cfDNA decreased in comparison to prior measurements, either at 30 days prior to or at progression. Conclusions: In almost all cases cfDNA could be detected in the blood of patients with RAS-mutated tumors which demonstrates the high sensitivity of the method. Data suggest a trend toward an increase of cfDNA during tumor progression. However, it is not clear why deviations, especially a decrease of VAF in cfDNA occurred in some patients prior to or at tumor progression. Currently unknown tumor-biological or therapy-associated factors might contribute to this finding. Further investigation is planned.
Dual role of TGFBR1 as a modifier of colorectal cancer risk.
Authors: Pennison Michael J. , Zeng Qinghua , Buckhaults Phillip J. , Kaklamani Virginia G. , Lindor Noralane M. , Hopper John L. , Le Marchand Loic , Gallinger Steven , Newcomb Polly A. , Haile Robert W. , Baron John A. , Stram Daniel O. , Zhang Hong-Tao , Manne Upender , Edberg Jeffrey C. , Kimberly Robert P. , Xu Jianfeng , Zhang Kui , Yi Nengjun , Pasche Boris 
Published: February 2017

600 Background: Experimental and clinical evidence suggests that constitutively decreased Transforming Growth Factor Beta type I receptor (TGFBR1) signaling predisposes to colorectal cancer (CRC) development. However, associations between TGFBR1 variants and CRC risk in case-control studies have been inconsistent. Methods: We utilized 1,043 CRC cases and their 1,627 unaffected sibling controls obtained from the Colon Cancer Family Registry (C-CFR). Individuals were genotyped for twelve TGFBR1 haplotype tagging SNPs. SNPs associated with CRC risk were validated in 261 CRC cases and 531 controls of African American ancestry and 990 CRC cases and 3,427 controls of Han Chinese ancestry. Validated SNPs were functionally characterized with respect to TGFBR1 expression and TGF-β signaling. Results: The TGFBR1 rs7034462-TT genotype was associated with CRC risk in C-CFR participants (OR 3.80[1.46-9.85]) and African Americans (OR 8.16[2.07-32.08]) (see Table). The TT genotype was associated with stage III and stage IV at diagnosis in C-CFR participants (OR 2.99[1.15-7.81] and OR 9.38[1.54-57.28]) and African Americans (OR 5.89[1.15-30.02] and OR 12.75[1.88-86.33]), respectively. The rs7034462-CT genotype was associated with decreased risk for CRC in African Americans (OR 0.55[0.31-0.99]) and Han Chinese (OR 0.67[0.48-0.95]) (see table). Cells carrying the rs7034462-TT genotype exhibited decreased constitutive TGFBR1 expression, increased SMAD7 expression, and decreased TGF-β signaling. Conclusions: The TGFBR1 rs7034462-T allele has dual, opposite roles with respect to CRC risk. The rare rs7034462-TT genotype is a moderate penetrance genotype associated with risk for CRC and advanced stage disease at diagnosis. In contrast, the common rs7034462-CT genotype is associated with decreased CRC risk. [Table: see text]
Preoperative treatment to modify the immune microenvironnement of liver colorectal metastases.
Authors: Van Den Eynde Marc , Mlecnik Bernhard , Machiels Jean-Pascal H. , Debetancourt Daphne , Bindea Gabriela , Pairet Geraldine , Jouret-Mourin Anne , Sempoux Christine , Gigot Jean Francois , Hubert Catherine , Humblet Yves , Carrasco Javier , Haicheur Nacilla , Marliot Florence , Pages Franck , Galon Jerome 
Published: February 2017

602 Background: We previously reported that an adaptive Th1 immune response (CD3/CD8/CD45RO T-cells) observed in resected primary colorectal tumor and liver colorectal metastases (LCM) is an important prognostic factor. B and FoxP3 regulatory lymphocytes participate to the modulation of this response. We aimed to investigate whether the preoperative treatments influenced the quality and the density of the immune infiltrates previously reported in the LCM. Methods: We used a cohort of metastatic colorectal patients (n=107) engaged for curative liver surgery with available FFPE blocks for all resected LCM to confirm the prognostic impact of the immune response. Among this cohort of 338 LCMs, 46 were completely resected after chemotherapy (CT) alone, 130 after CT + anti-VEGF, 118 after CT + anti-EGFR and 44 after surgery alone. LCMs were analyzed for histological response according the Tumor Regression Grade (TRG) and regrouped as Response (R, TRG1-3) or No Response (NR, TRG4-5). The density of CD3+ (T-cells), CD8+ (cytotoxic), CD45RO+ (memory), CD20+ (B-cells) and FoxP3+ (regulatory) in the core (CT) and invasive margin (IM) of all LCM was quantified on immunostained slides. The mean density value (CT/IM) was calculated for each marker with a dedicated image analysis software on whole-slide imaging. Comparisons were made using the Wilcoxon-Mann-Whitney test. Results: LCMs showing R (compared to NR and untreated LCM) were more frequently associated with a high immune infiltrate for CD3+ (CT: p<0.005; IM: p<0.05), CD8+ (CT: p<0.005; IM: p<0.005) and CD20+ (CT: p<0.05). Conversely, high FoxP3+ density in the CT and IM was related to NR and untreated LCMs (p<0.01). LCMs treated with an anti-EGFR therapy showed higher densities of CD3+ (CT: p<0.005; IM: p<0.01), CD8+ (CT: p<0.005), CD45RO+ (CT: p<0.005), CD20+ (CT: p<0.005) and FoxP3+ (CT: p<0.05; IM: p<0.005) compared to other treatments and untreated LCMs. Conclusions: Preoperative treatment modifies the LCM immune microenvironnement. LCMs with a histological response show a cytotoxic immune response (CD3+/CD8+) with associated B-cells (CD20+) and downregulated Tregs (FoxP3+). The use of an anti-EGFR therapy significantly increases immune infiltration in the CT.
Effect of DNA-PK dependent phosphorylation of topo-I-S10 on its rate of proteasomal degradation and CPT response.
Authors: Shah Ankur K. , Sachdew Vibhu , Taylor-Parker Julian , Tapan Umit , Tohme Yara Hamade , Malinkovich Yevgeniya , Tran Ben , Gibbs Peter , Salgia Ravi , White Forest M. , Parvin Jeffrey , Bharti Ajit 
Published: February 2017

606 Background: Topoisomerase I (topo-I) degradation by ubiquitin proteasomal pathway (UPP), in response to camptothecins (CPTs) is linked to the CPT response. But the mechanism is not understood. Methods: Improvised GST pull down experiments were performed to isolate topo-I interacting proteins which were analyzed by SDS-PAGE and identified by mass spectrometry. DNA-PKcs mediated phosphorylation of GST-topo-I was analyzed by SDS-PAGE and autoradiography. Immunoblot analysis of anti-BRCA1 precipitates from HCT15 cell lines was performed. GST-topo-I phosphorylated by DNA-PKcs, was incubated with the BRCA1/BARD1 heterodimer in the presence of E1, UbCH5c and ATP in ubiquitination buffer. Four triple negative breast cancer cell lines were selected to compare the degree of topo-I degradation. Colorectal cancer (CRC) tissues were immunostained with 1C1.H5.H7 to determine topo-I-pS10 level and identify a correlation between the topo-I-pS10 level and CPT response. Results: We isolated a topo-I interacting protein complex and determined that Ku70/Ku80/DNA-PKcs complex associates with topo-I, and DNA-PKcs phosphorylates topo-I at Serine10. We showed that cells with higher topo-I-pS10 level rapidly degrade topo-I and are CPT resistant. The cells with non-detectable level of topo-I-pS10 fail to degrade topo-I and are CPT sensitive. Retrospective study with 48 CRC tissues immuno-stained with anti-topo-I-pS10 supported our cell line data. Percent DAB positive nuclei demonstrated statistically significant differential staining between CPT responders and non-responders, and ROC analysis confirmed that nearly all non-responders exhibit >35% positive nuclei, indicating that topo-I-pS10 level determines topo-I degradation and a potential predictive biomarker for CPT response. Conclusions: Colorectal cancers are treated with topo-I inhibitors, but the response rate is low. We have shown that topo-I degradation rate is linked to CPT response. We have determined that high topo-I-pS10 level is indicative of rapid topoI degradation and can serve as a biomarker for CPT resistance.
The impact of oral anticancer drugs on recurrence pattern after adjuvant chemotherapy in colorectal cancer patients.
Authors: Shimada Takehiro , Tsuruta Masashi , Hasegawa Hirotoshi , Okabayashi Koji , Seishima Ryo , Takahashi Hidena , Matsui Shimpei , Yamada Toru , Kondo Takayuki , Matsuda Mutsuhito , Yahagi Masashi , Yoshikawa Yusuke , Asada Yusuke , Sugiura Kiyoaki , Suzuki Yoshiyuki , Tajima Yuki , Nakadai Junpei , Kitagawa Yuko 
Published: February 2017

610 Background: In colorectal cancer (CRC), liver metastasis is one of the critical prognostic factors. However, its pathogenesis still remains unclear. “Stream line theory” is known to be in the liver, where the bloodstream from superior and inferior mesenteric vein make a laminar flow in portal vein, and pour into the right and left hepatic vein, respectively. Recently we have elucidated that stream line theory might affect the pattern of liver metastasis in CRC. We anticipated that the way how to administrate anticancer drug also influence on the effectiveness for liver metastasis in CRC, namely oral administrated drug is absorbed in small intestine and delivered to the right lobe in the liver mainly. We hypothesized that oral intake of anticancer drug suppressed especially recurrence in the right hepatic lobe. The aim of this study is to elucidate the specific pattern of recurrence after curative resection of CRC depending on the way to administrate anticancer drug as adjuvant chemotherapy (AC). Methods: Between January 2003 and September 2013, of 365 patients who were received AC following R0 excision of primary CRC in our institution, we analyzed 94 patients who were diagnosed recurrence. They were divided into 2 groups: 64 patients were received only oral anticancer drug as AC following curative resection of CRC (PO group), and the others are distributed into IV group. A comparative review of clinicohistopathological factors and recurrence pattern were statistically performed. Results: Preoperative CEA levels were significantly higher in IV group than PO group (3.3 vs. 7.6, p=0.03). No other difference was observed in background factor of 2 groups. Comparing the recurrence pattern, 11 (17.2%) were diagnosed recurrence in the right hepatic lobe in PO group while 11 (36.7%) in IV group. There was significantly lower incidence of recurrence in the right hepatic lobe in PO group comparing with IV group (HR=2.79, 95% CI=0.92 to 8.37; p=0.04). Conclusions: AC by oral anticancer drug might specifically suppressed recurrence in the right hepatic lobe behind “stream line theory” in which oral anticancer drug is absorbed in small intestine and mainly drained into the right hepatic lobe.
Notch expression and bevacizumab efficacy in colorectal cancer patients.
Authors: Negri Francesca , Sala Roberto , Bozzetti Cecilia , Crafa Pellegrino , Lagrasta Costanza , Pedrazzi Giuseppe , Gardini Giorgio , Tamagnini Ione , Bisagni Alessandra , Azzoni Cinzia , Bottarelli Lorena , Romano Ida , Porzio Rosa , Bacchini Gian Paolo , Paties Carlo , Tomasello Gianluca , Leonardi Francesco , Ardizzoni Andrea , Pinto Carmine 
Published: February 2017

612 Background: Antiangiogenic therapies represent a well established additional treatment to standard chemotherapy, nevertheless no markers are available to suggest a successful outcome linked to the use of bevacizumab. To investigate potential mechanisms of resistance to angiogenesis inhibitor bevacizumab, Notch expression was correlated with response and survival in a series of bevacizumab treated advanced colorectal cancer (CRC) patients. Methods: Notch expression was evaluated by immunohistochemistry (IHC) on 65 primary CRC enrolled within 6 randomized clinical trials assessing first-line bevacizumab plus chemotherapy. Notch IHC was conducted using a polyclonal antibody to Cleaved Notch1 (Cell Signaling Technology, Danvers, MA). Notch expression was scored based on intensity of staining (0: none, 1+: weak, 2+: moderate; 3+: strong) and on percentage of immunostained cells. A control series of 21 advanced CRC treated with chemotherapy alone was also examined. Results: Notch positivity was localized to the cytoplasm or nucleus of malignant epithelial cells. In all, 11 of 61 (18%) evaluable primary tumours had a high Notch expression (IHC 3+). Six of the 11 cases (55%) with high Notch expression (IHC 3+) experienced progressive disease compared with 5 of 50 (10%) low Notch expression cases (IHC 0 1+ 2+) (p = 0.003). A high Notch expression also demonstrated an inferior median PFS (4.9 vs. 12.1 months; HR = 2.51; 95% CI, 0.96 to 6.58; p = 0.007) and OS (19.3 vs. 30.4 months; HR = 2.21; 95% CI, 0.79 to 6.15; p = 0.039) compared with low Notch expression cases. When the groups were further analyzed considering VEGF expression, the best outcome was found in low Notch (IHC 0 1 +) and high VEGF expressing tumors (IHC 2+ 3+) (response rate 9 of 11, 82 % vs. 1 of 5, 20%, in patients with high levels of Notch and VEGF expression, respectively). No correlation was found between Notch expression and clinical response in the series of patients treated with chemotherapy without bevacizumab. Conclusions: Notwithstanding the limited power of the present analysis, these data seem to suggest that low Notch expression might be a marker for successful bevacizumab treatment.