bluuebunny/crossref_metadata_2025 huggingface dataset:
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.
Multicenter trial for assessing cytokine promoter gene polymorphism as a predictive parameter of PSK responder for curatively resected stage II or III colorectal cancer.
Authors: Yoshino Shigefumi , Takumi Furuya , Sakata Koichiro , Shimizu Ryoichi , Okayama Naoko , Hazama Shoichi , Oka Masaaki
Published: February 2017

619

Background: Polysaccharide-K (PSK), a protein-bound polysaccharide extracted from the mycelia of Coriolus versicolor, is an immunomodulator widely used in colorectal cancer in Japan. PSK has immunological actions including enhancement or inhibition of cytokine production. It has been reported that levels of cytokine production are influenced by polymorphisms in the promoters of cytokine genes. We hypothesized that cytokine promoter gene polymorphisms may be responsible for genetic susceptibilities to immunological effect of PSK. Methods: This is a multicenter prospective trial. One hundred and ten patients with stage II or III colorectal cancer were enrolled. All patients received adjuvant immnochemotherapy after curative resection using UFT (stage II) or UFT/LV (stage III) combined with PSK (3.0 g/day, p.o.) for 1 year. Post-operative survey of recurrence was followed with CT scan at 6-month intervals during the first 2 years after surgery and at 1-year intervals thereafter until 5 year after surgery. DNA was extracted from peripheral blood cells in all patients. The following polymorphisms of the patients were genotyped: TNF-α-1031T/C, IL-1ß-511C/T, IL-6-634C/G, IL-10-819T/C. Results: Twenty (6 in stage II, 14 in stage III) out of 110 patients showed recurrence after more than 5 years survey. Fifteen out of 74 patients with TNF-α-1031 TT genotype and 5 out of 36 with TNF-α-1031 CC or CT showed disease recurrence. Five out of 39 patients with IL-1ß-511 CC and 15 out of 71 with IL-1ß-511 CT or TT showed disease recurrence. Eleven out of 61 patients with IL-6-634 CC and 9 out of 49 with IL-6-634 CG or GG showed disease recurrence. No association between genotype frequency and disease recurrence was observed for TNF-α, IL-1ß, IL-6. Fifteen out of 61 patients with IL-10-819 CC or CT genotype showed disease recurrence, whereas only 5 out of 49 with IL-10-819 TT showed tumor recurrence (p=0.052). Especially, significant association with disease recurrence was found in stage III patients (12/30 vs 2/17, p=0.042). Conclusions: It is suggested that colorectal cancer patients with IL-10-819 TT genotype could be PSK responder after curative resection.
Macrophage polarization related gene variants to predict clinical outcome in metastatic colorectal cancer (mCRC) patients (pts) treated with bevacizumab (bev) in combination with FOLFIRI.
Authors: Sunakawa Yu , Stintzing Sebastian , Heinemann Volker , Loupakis Fotios , Cremolini Chiara , Cao Shu , Yang Dongyun , Zhang Wu , Parekh Anish , Yamauchi Shinichi , Ning Yan , Stremitzer Stefan , Matsusaka Satoshi , Mendez Angela , El-Khoueiry Rita Elie , Lenz Heinz-Josef
Published: February 2017

621

Background: Macrophages, particularly M2, enhance tumor progression by stimulation of angiogenesis, acceleration of tumor invasion, and suppression of immunosurveillance. CCL2 promotes the polarization into M2 whereas histidine-rich glycoprotein (HRG) redirects M2 to M1. We hypothesized that single nucleotide polymorphisms (SNPs) in CCL2 and HRG may predict clinical outcome in mCRC pts treated with anti-VEGF drug in combination with chemotherapy. Methods: This study enrolled 4 pts cohorts treated with antibody drug plus FOLFIRI in prospective trials, A: KRAS exon 2 wild-type (KRAS wt) pts (n=84), B: mutant-type pts (n=89) from FOLFIRI-arm of TRIBE trial (NCT00719797), C: bev-arm (n=295), D: cetuximab-arm (n=297), served as negative control arm, from KRAS wt pts of FIRE3 trial (NCT00433927). Functionally significant SNPs, CCL2 rs4586, HRG rs9898, and HRGrs2228243, were analyzed by PCR-based direct sequencing for associations with response rate (RR), progression-free survival (PFS), and overall survival (OS) by uni- and multivariable analyses. Results: The minor G allele of HRG rs2228243 was significantly associated with better PFS, but not RR and OS, than the A/A genotype in both uni- and multivariable analyses (10.8 vs 9.3 months. HR: 0.62, p=0.048; HR: 0.57, p=0.040, respectively) in the cohort B while it was not associated in the other KRAS wt cohorts. In the cohort C, the minor C allele of CCL2 rs4586 correlated with significantly shorter OS, but not RR and PFS, in both uni- and multivariable analyses (22.3 vs 28.4 months. HR: 1.61, p=0.003; HR: 1.63, p=0.004, respectively). No significant association of CCL2 rs4586 was seen in the other cohorts although pts with the C allele of CCL2rs4586 had a negative marginal significant association in PFS and a shorter OS than pts with the T/T genotype (22.0 vs 30.8 months) in the cohort A. Conclusions: Our study provides evidence that SNPs in CCL2 and HRG may serve as a predictive or prognostic marker in mCRC pts treated with bev plus FOLFIRI. Our results also suggest that the impact of the SNPs on outcome may differ according to KRAS status. Prospective validation of this study is warranted.
Genetic analysis of colorectal cancers in young patients.
Authors: Abbott Andrea M. , Kothari Nishi , Teer Jamie K. , Srikumar Thejal , Kim Richard D. , Reed Damon R. , Shibata David
Published: February 2017

632

Background: The incidence of colorectal cancer (CRC) is increasing in adults <50 years old and these cases may be associated with a worse prognosis. Despite our growing understanding of genetic conditions, a substantial number of CRC in young patients are classified as sporadic but may harbor unique molecular changes. We sought to compare profiles of genetic alterations between young and old patients who lack defects in known hereditary cancer genes. Methods: 283 CRC cases diagnosed between 1998 and 2010 were analyzed by targeted exome sequencing using the Illumina NGS platform with 50-100x coverage. Filtering of normal variants was performed using 1000 Genomes to enrich for somatic mutations which were then limited to those predicted to alter protein sequence. The younger and older cohorts were defined as ≤45 and ≥65 years old at diagnosis, respectively. Patients found to be MSI-high (n= 2 young, 50 old) or with a known hereditary syndrome (n=6) were excluded. For this preliminary screen, Fisher’s Exact test was used to detect differences in mutation frequencies. Results: A total of 195 older and 30 younger patients with median ages of 73 (range 65-93) and 42 (range 30-45) years respectively, were analyzed. We identified 57 genes with significant differential mutation frequencies. The top ten genes mutated with increased in frequency in the younger cohort are shown in Table 1. Conclusions: In this exploratory project, we identified mutations that occurred more frequently in younger CRC patients. Large scale validation of these findings and application of this approach may lead to novel screening and treatment strategies in younger patients [Table: see text]
Contribution of FOXC1 to the progression and metastasis and prognosis of human colon cancer.
Authors: Li Dawei , Li Qingguo , Zhuo Changhua , Xu Ye , Cai Sanjun
Published: February 2017

636

Background: Distant metastasis remains the most common causes to death of colon cancer. Thus it is crucial to identify the molecular markers associated with the progression and metastasis of this disease. Recent evidence for overexpression of FOXC1 in several types of human cancer suggests that it might play a key role in tumor biology. However, the clinical significance of FOXC1 signaling in human colon cancer pathogenesis remains unknown. Methods: We investigated FOXC1 expression in 203 cases of primary colon cancer and matched normal colon tissue and lymph node matastasis in a tissue array. The underlying mechanisms of altered FOXC1 expression and the impact of this altered expression on colon cancer growth and metastasis was explored both in vitro and in vivo. Results: We found elevated expression of FOXC1 protein in cancereous tissue and lymph node metastases than adjacent normal colonic tissues. Overexpression of FOXC1 was associated with higher clinical stage, T stage, lymph node metastasis and presence of distant metastasis. FOXC1 served as an independent prognostic marker whose expression levels correlated with poorer metastasis-free survival (MFS) and poorer overall survival (OS). A Cox proportional hazards model revealed that FOXC1 expression was an independent prognostic factor in multivariate analysis. Experimentally, FOXC1 silencing significantly inhibited the growth and metastasis of colon cancer cells in vitro and in vivo. FOXC1 transcriptionally regulates SNAIL1, contributing to epithelial-to-mesenchymal transition and metastasis in colon cancer cells. Conclusions: Dysregulated expression of FOXC1 may play a critical role in colon cancer progression and metastasis. Thus, FOXC1 may serve as a candidate prognostic biomarker and therapeutically targeted.
Radiologic and pathologic prognostic factors after neoadyuvant chemotherapy for T3 rectal cancer (RC): 3-year update GEMCAD 0801-trial.
Authors: Machado Isidro , Brown Gina , Estevan Rafael , Salud Antonieta , Gil Mabel , Montagut Clara , Busto Marcos , Safont Maria Jose , Maurel Joan , Ayuso Juan Ramón , Aparicio Jorge , Feliu Jaime , Vera Ruth , Alonso Vicente , Gallego Javier , Martin Martha , Pericay Carles , Ballesteros Eva , Santos Jesus , Fernandez-Martos Carlos
Published: February 2017

643

Background: Prognostic factors in RC treated only with neoadjuvant chemotherapy, have not been explored. We analized the prognostic value of clinical-radiological and pathological factors for disease-free, (DFS) and cumulative incidence of distant metastases (DM), for patients treated with preoperative capecitabine, oxaliplatin and bevacizumab (CAPOX-B) within our multicentre phase II GEMCAD 0801 trial. Methods: 46 patients were enrolled to evaluate safety and efficacy of neoadjuvant CAPOX-B followed by surgery. Results have been recently published (Oncologist 2014;19:1-2). Eligibility included baseline magnetic resonance (MR) showing a T3 tumour with mesorectal fascia potentially clear. Clinical, pathologic (ypN+, T or N downstaging, tumor regression grade [TRG], pathologic complete response [pCR]) and radiologic factors both at baseline (mr extramural venous invasion [EMVI]) and post neoadjuvant chemotherapy (ymr TRG, ymr lengh change [RECIST]), were analyzed. Univariate and multivariate analysis was performed. Results: With a median follow up of 36 months, fourteen patients experienced relapse (2 local, 11 distant, 1both). 3-year DFS was 69%. It was 95%/49% for T downstaging/no T downstaging (p=0.0009) and 95%/43% for mrEMVI positive/negative (p=0.0001), respectively. ymrTRG (Mandard) ranging from no regression, TRG 5, to complete response, TRG 1, p=0.0108 and ypN0/ypN+ (p=.02) were also significantly related to DFS in univariate analysis. The same factors and N-downstaging were also significant for cumulative incidence of DM. On Cox multivariate analysis, T downstaging and mrEMVI were the only independent prognostic factors for DM ( p=0.0363 and 0.0111 respectively) and DFS (p=0.0315 and 0.0277 respectively). Conclusions: T3 rectal cancer with MR detected EMVI positive and those without downstaging after neoadjuvant chemotherapy are associated with unfavourable prognosis. This suggests that future strategies for treatment intensification and surveillance could be based on EMVI and T staging. Clinical trial information: NCT00909987.
Genomic markers of panitumumab resistance including ERBB2/HER2 in a phase II study of KRAS wild-type (wt) metastatic colorectal cancer (mCRC).
Authors: Barry Garrett , Li-Chang Hector Hugo , Kennecke Hagen F.
Published: February 2017

646

Background: Epidermal Growth Factor Receptor Inhibitors (EGFRi) are effective agents in the treatment of mCRC but up to one third of KRAS wt tumors do not respond. The primary objective was to identify a gene expression signature associated with primary resistance to panitumumab (Pmab) monotherapy and identify genomic markers associated with resistance. Methods: Patients with previously treated codon 12/13 KRAS wt mCRC were administered Pmab 6mg/kg IV q2weeks in a prospective phase II study. The primary endpoint was tumor response by RECIST and patients with progressive disease 8 weeks after initial Pmab were classified as resistant. Gene expression analysis was performed using a Nanostring-based assay with candidate genes implied in EGFRi resistance expression patterns. Differentially expressed genes were identified by Significance Analysis of Microarrays. Further interrogation of the HER2 signaling pathway was performed by immunohistochemistry and in situ hybridization. Illumina-based targeted resequencing was conducted on 20 cancer-implicated genes. Results: Of 34 patients treated with Pmab, 11 (32%) had progressive disease at 8 weeks and were classified Pmab resistant. A 5-gene expression signature was significantly associated with Pmab resistance (p = 0.001), including ERBB2, MLPH, IRX3, c11orf9, and KLK6. HER2 protein was overexpressed in 4/11 resistant and 0/21 responding cases (p = 0.035). Two (2) resistant tumors had ERBB2 gene amplification only, and one demonstrated both ERBB2 amplification and mutation. A non-codon 12/13 RAS mutation occurred in one treatment-resistant patient and was mutually exclusive with ERBB2 mutation, amplification, or HER2 overexpression. Four BRAF mutations were detected, two occurring in resistant patients. Conclusions: In this prospective study, a 5-gene signature including ERBB2 was associated with primary resistance to single agent Pmab. A subgroup of patients had evidence of aberrant HER2/ERBB2 signaling by protein over-expression and gene amplification. Results suggest a significant portion of KRAS wt tumors unresponsive to EGFRi may be identified by gene signature analysis. Clinical trial information: NCT00853931.
Phenotypic differences among RAS mutational variations in colorectal cancer (CRC): Analysis of 1,001 patients in single institute.
Authors: Shinozaki Eiji , Miki Yoshio , Ueno Masashi , Igarashi Masahiro , Suenaga Mitsukuni , Chin Keisho , Ozaka Masato , Ogura Mariko , Takahari Daisuke , Ichimura Takashi , Wakatsuki Takeru , Mizunuma Nobuyuki
Published: February 2017

649

Background: RAS mutation has been reported as a predictor of benefit from anti EGFR therapies in the recent sub-analysis of some large prospective trials, PRIME, FIRE-3 and so on. On the other hand, some in vitro experiments had identified the differences of capability of GTP bounding and colony formation according to KRAS mutational variations. It remains unknown whether there are phenotypic differences clinically among RAS mutational variations as potentially negative predictive factors for anti EGFR treatments. This study aimed to reveal the prevalence and the phenotypic differences of these variations in a large cohort. Methods: Consecutive patients of 1,001 CRC from January 2012 to October 2013 were analyzed in this study. Multiplex genotyping of EGFR pathway was performed on archival samples using Luminex Assay (MABGEN and GENOSEARCH Mu-PACK, MBL) for KRAS including codons 61,146 and NRAS. We examined the correlation among mutation variations; KRAS G12D, G12V, G13D and other exon2, exon 3 and 4 and NRAS, and clinical phenotypic features. Statistical analysis was conducted by chi-square test. Results: The incidence of mutations; KRAS G12D, G12V, G13D, other exon2, exon 3 and 4, and NRAS were 16.2%, 10.0%, 6.4%, 7.0%, 4.6% and 3.5%, respectively. So called all RAS mutations were occupied in 47% in this cohort. There was no significant difference of prevalence among mutational variations according to clinical feature as follows; clinical stage, gender, age. In histopathology, moderately differentiated type of G12D was more frequent compared with other type of mutations. The respective proportion of KRAS exon 3 and 4, and NRAS mutations for left sided primary was relatively higher than those for right sided, although the frequency of other mutations were higher in right side. Conclusions: According to our analysis there were not so large phenotypic differences among these RAS mutational variations except histology and localization of primary. Futher follow up of this cohort will provide more information about treatment outcomes in near future.
Are circulating tumor cells (CTCs) a feasible tool for predicting disease recurrence and survival in nonmetastatic (M0) colorectal cancer (CRC)?
Authors: Bergamo Francesca , Rossi Elisabetta , Pucciarelli Salvatore , Rossi Giovanna , Facchinetti Antonella , Battaglin Francesca , Toppan Paola , Fiduccia Pasquale , Amadori Alberto , Lonardi Sara , Zamarchi Rita , Zagonel Vittorina
Published: February 2017

650

Background: Based on the correlation between CTCs and clinical outcome of metastatic breast, prostatic and CRC cancer, CTCs might be an ideal model to investigate the individual risk to develop metastasis. On the other way, data on their significance in M0 CRC are still limited. In a previous study we investigated CRC cell death in the circulating compartment, finding high value of apoptotic CTCs (M30-positive) independently from the disease stage. Our aim was to evaluate the prognostic role of total and M30-positive CTCs in M0 CRC patients (pts). Methods: From January 2009 to December 2011, we prospectively enrolled 192 stage 0-III CRC pts (stage 0 identified rectal cancer pts with Tis or complete pathological response after neoadjuvant therapy). We used CellSearch assay to quantify total and M30-positive CTCs in peripheral blood of CRC pts candidate to curative resection at diagnosis, time of surgery and four months later. Multivariate survival analyses were conducted using a Cox-regression. Results: At baseline 81 out of 192 pts (43.3%) had at least 1 CTC/7.5 mL of peripheral blood, 40 pts (22.2%) had >2 cells and 20 pts (11.6%) had >3 cells. We found a weak association between OS and total CTCs number (p=0.069), a significant association with M30-positive CTCs (p=0.022) and no association between RFS and CTC status. Limiting the analysis to stage I-III (n=151), we found a strong association between OS and both CTCs number (p=0.021) and M30-positive cells (p=0.007). The multivariate analyses demonstrated a significant improvement of survival prediction adding baseline CTCs count to clinical and pathological characteristics (HR=1.51, 95% CI 1.16-1.96). Conclusions: We first demonstrated a significant association between OS and CTCs status in stage I-III CRC pts. This result is consistent with previous report in metastatic CRC. It highlights a potential advantage of CTCs count in stratifying CRC pts and identifying a subset of pts with a high metastatic risk that could benefit from an adjuvant therapy. Further studies are needed to explore this aspect, which might be particularly significant in stage II CRC where the indication for adjuvant therapy is still under debate.
The difference between the subjective and objective evaluation of oxaliplatin-induced vascular pain due to administer chemotherapy via peripheral vein.
Authors: Aisu Naoya , Yoshida Yoichiro , Yamada Teppei , Matsuoka Taisuke , Kojima Daibo , Tanimura Syu , Hoshino Seiichiro , Mogi Ai , Koganemaru Tomoko , Oda Mayumi , Fukuda Mahiru , Kiyomi Fumiaki , Noda Keita , Hirata Keiji , Tamura Kazuo , Yamashita Yuichi
Published: February 2017

651

Background: We have reported the safety and effectiveness of chemotherapy via median cubital vein for metastatic colorectal cancer without implantation of a central venous port. However, vascular pain (VP) occasionally requires switching of the drip infusion route during chemotherapy for the administration of oxaliplatin via the peripheral vein. VP and phlebitis induced by intravenous infusion of antineoplastic agents reduces the rate of completion or continuation of chemotherapy. We also reported that addition of dexamethasone to oxaliplatin drip infusion controlled the vascular pain caused by administration of oxaliplatin via the peripheral vein (ASCO-GI2012). Pain is currently evaluated using subjective methods such as the visual analog scale (VAS). However, because the assessment of pain can greatly vary depending on the mood and physical state of the patient at the time of assessment, it is best to objectively evaluate pain. Pain Vision PS-2100 (PV) is an analytical instrument that was designed to quantitatively and objectively assess sense perception and nociception in a patient. Although it is used in the field of anesthesiology, there have been no reports concerning it for the assessment of oxaliplatin-induced VP. Methods: The present study examined the correlation of subjective and objective assessment results using VAS and PV, respectively, for cases of oxaliplatin-induced VP. Subjects comprised 57 patients with colorectal cancer who underwent chemotherapy at the Fukuoka University. Results: Both VAS and PV assessments of PN were performed 162 times in total, and partial correlation coefficient analysis adjusted by subject and gender. The VAS and PV mean values of VP were 24.8 (0–100) and 44.5 (0–596), respectively. The partial correlation coefficient was 0.350 (p= 0.0002). Conclusions: Although both assessments evaluated the same events, no strong correlation was observed between the results and a weak correlation was observed between VAS and PV. These results suggest that because VAS and PV each measure different factors, both subjective and objective assessments of drugs designed to ameliorate oxaliplatin-induced VP are necessary.
Highly sensitive quantitative detection of circulating tumor DNA in urine and plasma from advanced colorectal cancer patients in aid of early diagnosis of clinically relevant KRAS mutations.
Authors: Poole Jason C. , Vibat Cecile Rose T. , Benesova Lucie , Belsanova Barbora , Hancock Saege , Lu Timothy , Erlander Mark , Minarik Marek
Published: February 2017

654

Background: Acquisition of point mutations in KRAS gene is causally associated with the onset of development of a resistance to anti-EGFR therapy in colorectal cancer. Newly acquired KRAS mutations can be detected in blood plasma months before radiographic detection. The objective of this study was to demonstrate feasibility of an ultrasensitive non-invasive method for detection of KRAS mutations in urine and plasma of patients with advanced colorectal cancer. Methods: Archived, matched urine and plasma samples (stored between 3-5 years prior to ctDNA extraction) from 20 treatment naïve, advanced stage cancer patients with known tumor tissue KRAS mutations determined by an accredited clinical laboratory, were used in a retrospective setting for a blinded concordance study. KRAS status in urine and plasma was compared to that in tumor tissue in order to assess clinical sensitivity of the ctDNA assay. An ultrashort-amplicon (31bp) assay for KRAS mutation enrichment and detection in highly fragmented urinary and plasma ctDNA was developed. The assay detected 1 copy of KRASG12A/C/D/R/S/V or G13D mutant allele in a background of wild-type DNA with a verified analytical sensitivity of 0.007% (7 copies per ~100,000 genome equivalents). Results: In a pilot study of 20 advanced stage colorectal cancer patients, 15 of 16 evaluable archived urine samples (94%) had KRAS mutation that was concordant with tissue biopsy. Of 20 archived plasma samples evaluated, 19 (95%) displayed the KRAS mutation concordant with tumor tissue. Of 16 paired urine and plasma samples, 15 (94%) had concordant KRAS mutation calls. Conclusions: This study demonstrates high clinical sensitivity (≥94%) of concordant KRAS mutation detection between urine, plasma and tissue specimens from advanced colorectal cancer patients. Early detection and monitoring of acquired KRAS mutations in circulating tumor DNA, and in particular urinary ctDNA, opens the possibility of a new paradigm for a truly non-invasive method of individualized care for colorectal cancer patients.
Anal cancer outcomes in patients treated with intensity modulated versus 3-dimensional chemoradiotherapy.
Authors: Cooper Benjamin Thomas , Grew David , Bitterman Danielle , Sanfilippo Nicholas , Du Kevin Lee
Published: February 2017

662

Background: Combined chemoradiotherapy (CRT) has been successful in both tumor eradication and colostomy prevention in patients with squamous-cell carcinoma of the anal canal. Unfortunately, CRT can be toxic with high rates of acute gastrointestinal and skin toxicity. This can necessitate treatment interruptions, prolonging therapy, possibly leading to loss of local control. In RTOG 0529, intensity modulated radiation therapy (IMRT) decreased the rate of treatment interruption compared to historical controls. We aim to compare toxicity and outcomes in patients treated with CRT based on radiation technique. Methods: We retrospectively reviewed 107 consecutive patients, 39 HIV+, 68 HIV-, who underwent definitive CRT for anal cancer at a single institution between 2004 and 2013. Overall survival (OS), colostomy-free survival (CFS), local recurrence-free survival (LRFS) and distant metastasis-free survival (DMFS) were analyzed. Chi-square test was used to compare frequencies and t-test was used to compare means. Kaplan-Meier survival was calculated and differences were evaluated by Log-rank statistic. Results: Median follow-up was 15 months. Radiation technique was IMRT in 60% of patients with the remainder treated with 3-dimensional conformal radiation therapy (3D). Dose to the draining lymph nodes was higher in patients treated with IMRT (mean dose 40 Gy vs. 32 Gy, p < 0.001). Fewer patients had a greater than 10 day treatment break in IMRT cohort than the 3D cohort (21% vs. 43%, p = 0.028). Three-year OS (91% vs. 47%, p < 0.001) and DMFS (88% vs 64%, p= 0.033) were improved in patients treated with IMRT. There was no significant difference in acute GI or skin toxicity. There was no difference in stage, number of chemotherapy cycles and dose reductions, growth factor support, transfusion necessity, hospital admission, LRFS, sphincter function preservation, or CFS. Conclusions: In this cohort, patients treated with IMRT had better OS and DMFS than patients treated with 3D. Higher radiation doses to the draining lymph nodes and fewer prolonged treatment breaks may contribute to improved outcomes in patients treated with IMRT. Further studies are necessary to establish the etiology of this difference in outcomes.