bluuebunny/crossref_metadata_2025 huggingface dataset:
An improved method for detection of methylated circulating tumor DNA in colorectal cancer.
Authors: Boulter Nicky , Tevz Gregor , Yu Betty , Chan Michelle , Murray David , Symonds Erin L. , Young Graeme P. , Pedersen Susanne Kartin
Published: May 2019

e15120

Background: Assays detecting methylated circulating tumor DNA (ctDNA) hold promise for colorectal cancer (CRC) screening, but there is a need to develop more accurate assays for early stages. In this study, we examined modifications to COLVERA (an IKZF1 and BCAT1 methylation marker qPCR assay) that added a third biomarker (IRF4) as well as introducing double stranded detection. Methods: Bisulfite converted DNA extracted from plasma collected from subjects undergoing diagnosis for CRC was assayed using either a double-stranded (ds) specific multiplexed qPCR for BCAT1 and IKZF1 detection (Assay 1) and/or a single-stranded (ss) specific BCAT1/IRF4, ds-specific IKZF1 multiplexed qPCR (Assay 2). The unmodified COLVERA test (Clinical Genomics) was used for comparison. Detection of any marker was considered positive. Results: Assay 1 performance evaluated against COLVERA on bisulfite converted DNA from 1453 samples showed an improved sensitivity for CRC detection (93/134 (69.4%) vs 79/134 (59.0%) COLVERA, p = 0.02), but specificity was suboptimal (no neoplasia: 861/1023 (84.2%) vs 946/1023 (92.5%) COLVERA, p < 0.0001). The decrease in specificity resulted primarily from modifications in the BCAT1 assay component (1023 no neoplasia: 60 positive with ss- BCAT1 vs 146 positive with ds- BCAT1, p < 0.0001). Thus, a novel configuration, Assay 2 (ss- BCAT1, ss- IRF4 and ds- IKZF1) was compared to Assay 1 using 189 previously untested specimens. Assay 2 showed a notable improvement in specificity (99/104 (95.2%) vs 93/104 (89.4%) Assay 1; p = 0.0351). Both assays exhibited comparable sensitivities for CRC (41/49 (83.7%) vs 39/49 (79.6%), Assay 1, p = 0.3633). Assay 2 was positive in: adenomas, 9/36 (25.0%); Stage I, 3/7 (42.9%); Stage II, 17/19 (89.5%); Stage III, 6/6 (100%); Stage IV, 7/7 (100%) and 8/10 (80%) unstaged CRCs. Prior COLVERA sensitivity estimates were 9% adenoma, 41% Stage I and 76% Stage II. Conclusions: Targeting both of the non-complementary bisulfite converted strands of selective regions of interest markedly improve ctDNA assay sensitivity for cancer including early lesions while achieving good specificity. Prospective evaluation in a true CRC screening population is now underway. Clinical trial information: 12611000318987.
Mutation profiling of BRAF gene in Chinese patients with early or advanced colorectal cancer using comprehensive NGS panel.
Authors: Lin Canfeng , Jia Wanqiu , Sheng Wang , Zhang Lijiao , Zhang Feiran , Gu Yanhong , Zhu Lingjun , Chen Rongrong , Xia Xuefeng
Published: May 2019

e15124

Background: It has been reported that BRAF gene mutation occurs in approximately 10% of metastatic colorectal cancer (mCRC), and is associated with resistance to EGFR inhibitors. Moreover, patients with non-V600E BRAF mutations with a better prognosis than patients with V600E BRAF mutations in advanced colorectal cancer. However, little is known about mutation profiling of BRAF in early stage colorectal cancer. Methods: We reviewed mutation profiling of 1429 colorectal cancer samples in our institute from 2016 to 2018. Tissue biopsy and/or ctDNA samples were analyzed using hybridization capture-based NGS ER-Seq method, which enables simultaneously assess single-nucleotide variants, insertions/deletions, rearrangements, and somatic copy-number alterations at least 59 genes (range 59-1021 genes). Results: A total of 1429 samples from 1120 patients with CRC, included 86 early stage (Ⅰ-Ⅱ ), 966 advanced stage (Ⅲ -Ⅳ), and 377 samples with unknown stage. We identified 109 samples from 94 patients with BRAF gene mutation in the cohort (109/1429=7.63%), of which V600E and non-V600E were 62(4.34%) and 47 (3.29%), respectively. Among the 109 samples, there were 15 of early stage, 84 of advanced stage and 10 of unknown stages. BRAF mutation were more common in early CRC samples (15/86=17.44%) than in advanced CRC samples (84/966=8.7%, p = 0.007). Furthermore, 10 of the 15 early CRC samples yet 32 of the 84 mCRC samples had non-V600E were detected (66.67% vs 38.1%, p=0.039). In contrast to V600E mutation, some of the non-V600E variants were variants of unknown clinical significance (VUS) in this study (see Table). Conclusions: BRAF mutation is more common in early stage CRC, the higher prevalence of non-V600E BRAF mutations in early CRC may account for the better prognosis of early stage CRC. non-V600E BRAF mutation subtype. [Table: see text]
Radiomic profiles via machine learning to predict response, overall survival, and KRAS status in metastatic colorectal tumors treated with FOLFOX/bevacizumab/hydroxychloroquine.
Authors: Rathore Saima , O'Hara Mark H. , Karasic Thomas Benjamin , Redlinger Maryann , Rosen Mark Alan , Davatzikos Christos , O'Dwyer Peter
Published: May 2019

e15125

Background: Autophagy is a resistance mechanism to chemotherapy that is inhibited by hydroxychloroquine (HCQ). We have reported a phase II trial of FOLFOX/bevacizumab/HCQ in 28 evaluable patients with untreated metastatic colorectal cancer (mCRC). Overall response rate was 68%, with 11% complete response (CR) and 57% partial response (PR), while stable disease (SD) was seen in 30%. We hypothesize that analysis of CT imaging features via machine learning (ML) will enhance subtle yet important radiographic characteristics, and reveal imaging signatures determinant of outcome and mutational status. Methods: Baseline CT images were collected and 1265 quantitative imaging (QI) features extracted across all the metastatic sites – liver, lung, and lymph nodes – including descriptors of size, morphology, texture, and intensity. Cross-validated sequential feature selection coupled with support vector machine (SVM) was used to determine the most discriminative QI features for our integrative predictor of response and mutational status, and support vector regression (SVR) was used to derive imaging predictors of overall survival. The model predictions were compared with actual clinical results, including response, survival, and genomic aberrations. Results: Various QI features, primarily descriptive of texture and tumor volume, were determined as most important by the ML predictor. Using this signature, our predictor classified “Responder (PR+CR) vs Non-responder (SD)” with an accuracy of 85% [sensitivity(se) = 0.84, specificity(sp) = 0.71, AUC = 0.85]. The QI features were also able to detect KRAS and TP53 mutational status with an accuracy of 88% [se = 0.92, sp = 0.85, AUC = 0.87] and 86% [se = 0.80, sp = 0.90, AUC = 0.84], respectively. The SVM model predicted overall survival greater than the median (32 mo) with an accuracy of 86% [se = 0.93, sp = 0.79, AUC = 0.87]. The Pearson correlation coefficient between the SVR score and overall survival was estimated to be 0.73 (p < 0.0001). Conclusions: Radiomic analysis of baseline CT imaging features analyzed by ML yielded an imaging signature predictive of response, survival, and KRAS and TP53 mutational status. If validated in a larger clinical data set, machine learning may offer a predictive biomarker to aid clinical decision making for mCRC patients.
Colorectal cancer molecular subtypes by immunohistochemistry in a patient cohort from Nigeria.
Authors: Abdulkareem F Biade , Khramtsova Galina , Adedokun Babatunde , Rotimi Olorunda , Abudu Kunle , Badmos Kabir , Ibraheem Abiola Falilat , Khramtsov Andrey , Sveen Lise , Hurley Ian , Masaya Hattori , Huo Dezheng , Olopade Olufunmilayo I.
Published: May 2019

e15145

Background: Colorectal cancer (CRC) classificationusesclinico-pathological characteristics. However, patients with similar histologic features and tumor stages have different clinical outcomes and drug responses. Studies of CRCmolecular heterogeneity have used genome-wide gene-expression data to classifypatients into four consensus molecular subtypes (CMS), but the complexity of analysis has limited its use in practice. Methods: With IRB approval, tissue microarray (TMA) blocks were constructed, IHC was performed and semi-quantitative scores completed. For the MMR proteins, MLH1 (Pierce), MSH2 (LifeTechnologies), MSH6 (Novex), PMS2 (Pierce) antibodies were used. Three markers were selected from transcriptomic analysis for IHC: CDX2 (LifeTechnologies), HTR2B (LifeTechnologies), and ZEB1 (eBioscience). Ki-67 (DAKO) proliferation rate was used as anequivalent marker for β-catenin activation.The CMS1 subtype was first separated using MSI/MSS status. A panel of CDX2, HTR2B, and ZEB1was then used to distinguish between CMS4 and CMS2/CMS3 subtypes, while Ki-67 was used to separate CMS2 from CMS3 subtype. Results: Of the total evaluable 75 CRC cases(mean age 44.8 years (SD = 16.1), 38% were <40 years old, and 60% were males. Of tumors with data on tumor grade (n=66), 62% were well differentiated. 59 patients (79%) had MSS, and the remaining16(21%) had MSI (i.e. CMS1). 22 (29%) patients with increased ZEB1 and lower CDX2 expression were classified as CMS4. 37 (49%) displayed increased CDX2 and lower ZEB1 expression and were classified as CMS2 (n=24) or CMS3 (n=13). HTR2B was universally expressed, but a higher median HTR2B score of 3 was observed in CMS4 cases compared to 1 for other subtypes (p=0.0001). The CMS4 subtype was significantly more likely to occur among young patients (p<0.001): in patients < 40 years, 70% of patients were CMS4 subtype, while in patients > 40, 7% were CMS4 subtype. Of the left-sided cancers, only 5% were CMS1 subtype, while CMS1 composed 75% of right-sided cancers (p<0.001). Conclusions: Using TMAs and a panel of antibodies, we show an association of a poorer prognostic subtype (CMS4) with younger age at diagnosis. IHC-based classification and screening could be a cost effective prognostic and predictive tool for Nigerian reference laboratories.
Machine learning for CT texture analysis in chemosensitivity of colorectal liver metastases: Initial results.
Authors: Miyamoto Yuji , Nakaura Takeshi , Hiyoshi Yukiharu , Baba Hideo
Published: May 2019

e15148

Background: Despite advances in cancer treatment over the last decades, more efficacious biomarkers are needed in patients with metastatic colorectal cancer. Several studies have reported that CT texture analysis is a useful prognostic biomarker for patients with colorectal cancer liver metastases (CRLM), however, little study has been done to explore those efficacies using machine learning methods. The present study aimed to evaluate the clinical efficacy of CT texture analysis using machine learning methods as a predictive marker of systemic chemotherapy in patients with CRLM. Methods: Sixty-four patients with CRLM who received first-line chemotherapy were included. Texture analysis was performed on 92 features (First Order Statistics, Gray Level Cooccurrence Matrix, Gray Level Run Length Matrix, Gray Level Size Zone Matrix, Neighbouring Gray Tone Difference Matrix and Gray Level Dependence Matrix) using CT within 1 month before treatment. We evaluated the association between those features and chemotherapeutic response by RECIST (CR+PR vs. SD+PD+NE). We performed eXtreme gradient boost (XGBoost) as a machine learning method to predict the chemotherapeutic response and used the receiver operating characteristic curves to evaluate this prediction model. Results: Main characteristics were the following: male/female = 36/28; median age = 63.5. Patients were treated with oxaliplatin-based chemotherapy (80% of patients), bevacizumab (77%) and anti-EGFR antibody (23%). Thirty-nine patients had confirmed responders, for an overall response rate of 61%, whereas 25 patients (39%) were classified as non-responders (CR: PR: SD: PD: NE = 0: 39: 20: 4: 1). The area under curve of this prediction model was 0.771. Conclusions: We confirmed that CT texture analysis using machine learning for CRLM was feasible. Further analyses are ongoing.
Prognostic impact of mutation profiling in Chinese patients with colorectal liver metastases.
Authors: Wang Kun , Yang Yadong , Zhang Xiaoyu , Li Juan , Bao Quan , Wang Hongwei , Jin Kemin , Liu Ming , Liu Wei , Yan Xiaoluan , Wang Lijun , Da Xu , Xing Baocai
Published: May 2019

e15157

Background: To investigate mutation profiling in Chinese patients with colorectal liver metastases and explore new prognostic biomarkers. Methods: We investigated 197 patients who underwent surgery for colorectal liver metastases at Beijing Cancer Hospital between January 2015 and February 2017. Tumor DNA was isolated from FFPE tissues from liver metastatic lesions. Mutation spectrum was obtained by targeted deep sequencing of 620 selected genes. Tumor mutational burden (TMB) was defined as the number of non-synonymous mutations per 1Mbp and divided into quartiles. MSI status was tested with next-generation sequencing. Studies of impacts of mutation status on disease-free survival (DFS) and overall survival (OS) were performed with Kaplan-Meier analysis. Results: Mutation significance analysis confirmed enrichment in colorectal cancer driver gene mutations of TP53, APC, KRAS, SMAD4, SOX9, FBXW7 and PIK3CA in 72%, 61%, 29%, 14%, 10%, 10% and 9% of liver metastases, with no BRAF mutation found in any of these patients. Additionally, we observed frequent gene-level copy number variations, including amplifications of MYC, BRCA2, ERBB2, CCND2 and deletion of APC. Among 197 patients, only three showed MSI-high, who also exhibited high TMBs. Moreover, KRAS, MXRA5, and SETD2 mutations were significantly associated with shorter DFS ( KRAS mt vs wt: 8 months vs 11 months, p = 0.0086; MXRA5 mt vs wt: 6 months vs 11 months, p = 0.0178; SETD2 mt vs wt: 8 months vs 11 months, p = 0.0381), but KMT2A mutation associated with longer DFS (mt vs wt: 31.5 months vs 10 months, p = 0.0409). SPEN, GNAS, CTNNB1, SETD2 and FANCM mutations were significantly associated with poor OS ( SPEN mt vs wt: 23 months vs 30 months, p = 0.0047; GNAS mt vs wt: 24.5 months vs 30 months, p = 0.0386; CTNNB1 mt vs wt: 20 months vs 30 months, p = 0.0001; SETD2 mt vs wt: 20 months vs 30 months, p = 0.0026; FANCM mt vs wt: 13 months vs 30 months, p = 0.0007). Conclusions: Our study has found a novel panel of prognostic biomarkers in colorectal liver metastases. The clinical significance of mutations of MXRA5, SETD2, KMT2A, SPEN, GNAS, CTNNB1, FANCM needs to be confirmed on a larger scale.
Prognostic nutritional index and colorectal cancer deaths: A retrospective cohort study in Chinese patients with locally advanced colorectal cancer.
Authors: Wang Wenling
Published: May 2019

e15160

Background: The nutrition and immune status-based scoring system PNI was originally used to assess the nutritional status and surgical risk of surgical patients before surgery. Subsequent studies have shown that PNI is an important prognostic indicator for some malignant tumors, including hepatocellular carcinoma, pancreatic cancer, laryngeal cancer, renal cell carcinoma and gastric cancer. We speculated that since PNI is associated with the prognosis of other malignancies, is it also relevant in colorectal cancer? In view of the obvious increase in the incidence of colorectal cancer in China, and the clinical progress of colorectal cancer in the entire colorectal cancer accounted for up to 70% and current studies on PNI and local advanced colorectal cancer prognosis association study in Chinese is limited. In the present study, our primary objective was to explore whether PNI evaluated at baseline was independently associated with colorectal cancer deaths after controlling other potential confounders. Methods: Through a retrospective cohort study, we analyzed the clinical data of 400 patients with pathologically confirmed colorectal cancer and radical resection of colorectal cancer from January 2013 to April 2018.The exposure variable was PNI at baseline, the outcome variable was death, and other covariates of this study include: Continuous variables: age, platelet count, tumor markers (CEA, CA199, CA724, CA242).Categorical variables: gender, survival status, tumor location, histological type, degree of differentiation, TNM staging, and postoperative treatment. Results: Fully-adjusted model demonstrated that PNI was not associated with deaths of local advanced colorectal cancer and radical resection of colorectal cancer (HR = 0.88; 95% CI, 0.78-1.00). By applying the cubic spline function to the model, we found that the relationship between PNI and death is nonlinear.Two-piecewise cox model suggested that On the left side of the inflection point (41.35), PNI per 1 change is associated with the 68% decrease of risk of deaths (HR = 0.32; 95% CI, 0.13-0.79), to the right of the inflection point, PNI and death The association is not detected. Conclusions: PNI and deaths are nonlinearity, and within a certain range (PNI = 26-41), PNI is an independent protective factor for death risk.
Validation of neoadjuvant rectal cancer(NAR) score as a surrogate endpoint for disease free survival in Chinese FOWARC study.
Authors: Deng Yanhong , Cai Yue , Zhang Jianwei , Hu Huabin , Ling Jiayu , Wu Zehua , Wang Jianping
Published: May 2019

e15162

Background: NAR score is considered as a surrogate endpoint for survival in patients with locally advanced rectal cancer who received neoadjuvant treatment. Here we validate it in Chinese FOWARC study, especially in the chemotherapy alone arm. Methods: NAR score is calculated based on cT, ypT, ypN as reported for patients who received surgery in FOWARC study. NAR score is correlated to DFS with COX regression and Kaplan-Meier analysis. Results: 451 patients had surgery, 147 in FU-RT group, 152 in FOLFOX-RT arm and 152 in FOLFOX arm. NAR score is a significant prognostic factor in COX regression (P=0.000, HR 1.034, 95% CI 1.024-1.045) and in FOLFOX alone group (p=0.001, HR 1.030, 95% CI 1.013-1.047). When NAR score is quartile to <5, 5-10, 10-15,>15, the survival of patients is listed below, and similar to pathological staging. Conclusions: NAR score can be used as a surrogate endpoint for disease free survival Chinese patients. But it is not superior to pathological stage, especially in patients who received chemotherapy alone. [Table: see text]
Nomogram incorporated the number of adjuvant chemotherapy cycles for predicting the prognosis in stage II-III rectal cancer patients without neoadjuvant therapy.
Authors: Chen Weiwei , Wang Wenling
Published: May 2019

e15163

Background: Current recommendations for adjuvant chemotherapy in rectal cancer are based on the studies in colon cancer. However, it is now known that rectal cancer differs from colon cancer significantly regarding clinical course and biology. No RCTs in the TME era have evaluated the value of postoperative chemotherapy and are unlikely to be performed as neoadjuvant treatment has become a “gold standard” approach. However, we found that not all patients with locally advanced rectal cancer underwent neoadjuvant chemoradiotherapy before TME in real-world clinical practice in China. Whether the number of adjuvant chemotherapy cycles is significantly related to the prognosis of these patients deserves further study. Methods: A total of 246 patients with stage II-III rectal cancer from January 2013 to April 2018 were enrolled. All patients underwent surgery and had not received neoadjuvant therapy. The survival curve was drawn by the Kaplan-Meier method, and the log-rank method was used for statistical analysis. The Cox proportional hazard model was used for multivariate analysis to determine the independent prognostic factors. Then, MFP(Multiple Fractional Polynominal) and stepwiseAIC were used for variable selection. The R software was used to establish the nomogram. The bootstrap method was employed to internal verification. Concordance index(C-index) was applied to evaluate the predictive power of nomogram. Calibration curves were drawn to compare the 3-year overall survival rate predicted by nomogram and that of actual observation. Results: 87.8% of patients received adjuvant chemotherapy including oxaliplatin combined with fluorouracil or capecitabine. Univariate and multivariate analysis showed that the number of adjuvant chemotherapy cycles was independent prognostic factors. Patients who received more than 5 cycles of chemotherapy (HR = 0.09, 95%CI(0.01,0.80)) had a significantly better overall survival than patients with less than 5 cycles (HR = 0.33,95%CI(0.12,0.89)) or no chemotherapy (p < 0.05).Through MFP and the stepwiseAIC screening, a nomogram was established based on CEA, PLR, N, and the number of chemotherapy cycles, and the C-index of the model was 0.86. Conclusions: The number of adjuvant chemotherapy cycles is an independent prognostic factor in stage II-III rectal cancer patients without neoadjuvant therapy. Moreover, nomogram incorporated the number of chemotherapy cycles was accurate and visible.
Factors of local immunity in patients with rectal cancer after prolonged radiotherapy.
Authors: Kit Oleg Ivanovich , Snezhko Aleksandr V. , Zlatnik Elena Yu. , Novikova Inna A. , Al-haj Nabil , Bondarenko Elena S. , Vladimirova Liubov Yu
Published: May 2019

e15164

Background: Immunological study of the blood and tumor tissues was performed in patients with rectal cancer receiving neoadjuvant chemoradiotherapy. Methods: 30 patients with rectal cancer (13 women and 17 men aged 37-68 years with stage II-III adenocarcinomas G1-G3) were divided into groups according to results of DNA cytometry and their response to neoadjuvant treatment. When tumor proliferative activity was stable for 4 weeks of treatment, patients received surgery on time (group 1), while patients with verified inhibition of tumor proliferation (the index decrease by 1.5 times and more) continued treatment for 6-8 weeks and then were operated on (group 2). The immune status of patients (T, B, NK, DN, Tregs) was assessed during treatment. Homogenates of tumor tissue samples obtained during surgery were studied for the levels of lymphocytes (flow cytometry) and cytokines TNF-α, IL-1ß, IL-1RA, IL-6, IL-8, IFN-α, IFN-γ (ELISA); tumor proliferation index was assessed by DNA cytometry. Results were analyzed by Statistica 10.0 program. Results: The dynamics of parameters of the cellular immunity was different in patients of two groups. In group 1, percentage of T lymphocytes in blood decreased (from 66.7±3.3 to 50.4±1.6%), as well as their main subsets (CD4+ and CD8+ cells: from 33.6±2.7 to 27.0±1.7% and from 26.7±2.4 to 20.7±1.7% respectively). Patients of group 2 developed an increase in levels of NK cells from 10.1±1.2 to 15.3±2.2%, and levels of CD3+, CD4+ and CD8+ cells were significantly higher than in group 1: 35.0±1.8% for CD4+ and 28.3±2.9% for CD8+ (p < 0.05). The groups also differed in indices of local immunity: DN cells levels in group 2 were lower than in group 1 (5.8±1.0 vs. 18.4±5.4%) and CD4+ were higher (36.6±3.3 vs. 26.2±3.1%; p < 0.05). Patients of group 2 showed lower levels of IL-1ß, IL-6, IL-10, while IFNγ was elevated by 5.4 times, indicating a more favorable local cytokine status of the patients, compared to group 1. Conclusions: In rectal cancer patients with effect confirmed by DNA cytometry, prolongation of chemoradiotherapy to 6-8 weeks provides the formation of a more favorable immunological microenvironment of the tumor, and in such cases it is considered appropriate.
Patterns of local failure after definitive radiotherapy in patients with esophageal squamous cell carcinoma: Implications for the longitudinal clinical target volume design.
Authors: Cheng Bo , Meng Xue
Published: May 2019

e15504

Background: The correlation between the clinical target volume (CTV) margin and esophageal squamous cell carcinoma (ESCC) recurrence pattern is not completely clear. Our aim was to retrospectively evaluate the recurrence patterns in ESCC patients who received definitive radiotherapy (RT) with different CTVs, and determine the optimum longitudinalmargin. Methods: Patients with histologically proven ESCC (N = 134) were classified into the following three groups according to the initial expanding margin of CTV at the recurrence side: CTV1 with longitudinal margin ≥ 3cm (N = 73), CTV2 with 2-3cm (N = 37), and CTV3 with < 2cm (N = 24). The recurrence patterns were analyzed relative to the CTV margins, as well as the radiation dose, tumor stage, comorbidities etc. Results: The recurrence sites were significantly different between the CTV1 and CTV3 (p < 0.001), but not between the CTV1 and CTV2 groups (p = 0.052). In addition, 42 (91.3%) of the 46 patients that received low-dose (55-60Gy) RT showed recurrence at the site of the primary tumor, an obviously greater proportion compared to those who underwent a high-dose ( > 60Gy) treatment (77.3%, p = 0.018). Drinking was also a clear contributing factor (p = 0.025). Age, smoking, comorbidities,tumor location, stage and treatment mode were not significant prognostic factors for ESCC recurrence. Conclusions: CTV with a 2-3cm longitudinal expansion to the gross tumor volume (GTV) is acceptable for radical RT for ESCC, and recurrence at the primary tumor site is associated with low radiation dose ( < 60Gy).
Dosimetric and radiobiological comparison of external beam radiotherapy using simultaneous integrated boost technique for esophageal cancer in different location.
Authors: Wang Lu , Yu Jinming
Published: May 2019

e15505

Background: Based on dosimetry and radiobiology to compare treatment plans for esophageal cancer (EC) in different location using intensity modulated radiotherapy (IMRT), volumetric modulated arc radiotherapy (VMAT) and helical tomotherapy(HT) with simultaneous integrated boost (SIB) technique. Methods: A total of 20 patients including 5 cases respectively located in the cervix, upper, middle and lower thorax were generated for IMRT, VMAT and HT plans. The dose volume histogram statistics, conformity index (CI), homogeneity index (HI), tumor control probability (TCP) and normal tissues control probability (NTCP) were analyzed to evaluate treatment plans. Results: HT showed significantly improvement over IMRT and VMAT in terms of CI(0.93±0.03), HI(0.07±0.03) and TCP(88.08±0.82%) in cervical EC(p<0.05). IMRT greatly developed TCP(88.29±1.79%;85.11±0.79%), and offered superior CIs (0.87±0.04;0.90±0.01) and HIs(0.10±0.01; 0.06±0.01) compared with VMAT and HT in upper and middle thoracic EC(p<0.05). Meanwhile, the V30(33.30±6.49%), mean dose (2559.00±219.64cGy) and NTCP(0.50±0.61%) of heart for IMRT were significantly reduced than other two techniques in middle thoracic EC. Patients with lower thoracic EC yielded the similar CIs and HIs(all p>0.05) for the 3 techniques, but VMAT showed the lowest NTCP of lungs (0.01±0.01%) with improved TCP (84.84±1.13%). Conclusions: HT was a good option with little lung and heart involvement as it achieved superior dose conformality and uniformity. IMRT was a perfect strategy with large thoracic involvement. It significantly improved tumor local control and reduced heart dose and complications with acceptable dose to lungs. VMAT was preferred with a smaller target volume but surrounded by more heart and less lungs. Individually choosing optimal technique for EC in different location will be warranted.
A phase Ib study of oraxol in combination with ramucirumab in patients with gastric or esophageal cancers who failed previous chemotherapy.
Authors: Hung Yi-Ping , Chao Yee , Chen Ming Huang , Cutler David , Kramer Douglas , Chan Wing Kai , Kwan Rudolf , Hung Noelyn Anne , Hung Tak , Tenner Laura LaNiel
Published: May 2019

e15518

Background: Oraxol consists of oral paclitaxel (PTX) administered with the novel P-glycoprotein inhibitor HM30181A which enables the oral absorption of PTX. Ramucirumab (RAM) + intravenous weekly PTX is FDA approved 2nd line treatment of gastric cancer. Oraxol (oral PTX) 200mg/m2 days 1-3, weekly results in similar exposure to weekly 80/m2 intravenous PTX. This phase 1 clinical trial is to determine the maximum tolerated dose (MTD) of Oraxol + RAM. Methods: Eligible patients (pts) have gastric or esophageal cancers and failed prior fluoropyrimidine or platinum containing chemotherapies. Dose escalation followed the standard 3+3 design: Cohort 1: Oraxol 200mg/m2 days 1-3, weekly. Cohort 2: Oraxol 250mg/m2 days 1-3, weekly. Cohort 3: Oraxol 300mg/m2 days 1-3, weekly. Dose of RAM is standard 8 mg/kg IV every 2 weeks. Pts were monitored for dose limiting toxicity (DLT) by week 4. Adverse events (AEs) were assessed per CTCAE v4.03 and response by RECIST v1.1. Results: Cohort 1: One febrile neutropenia (DLT) observed in 6 pts. Partial response (PR) = 2/6, stable disease (SD) = 1/6 and progressive disease (PD) = 3/6 pts. Cohort 2: One grade-3 neutropenia with treatment delay (DLT) in 7 pts. PR = 3/6 and PD = 3/6 in 6 evaluable pts. Cohort 3: One febrile neutropenia (DLT) in 3 pts and study is ongoing. All pts had complete recovery of their DLT. PK showed increase of plasma PTX exposure with dose. Conclusions: The MTD for Oraxol has not been reached yet in combination with RAM. Preliminary results showed that Oraxol + RAM is tolerable and have encouraging anti-tumor activity in gastric or esophageal cancers pts who failed prior chemotherapies. Final results of this study will be presented. Clinical trial information: NCT02970539.
Investigation to metastasis of regional lymph node station and prediction to long-term survival following esophagectomy in thoracic esophageal cancer with stage T1 to T3.
Authors: Li Yin , Qin Jianjun , Li Xiaofei , Fu Xiangning , Liu Lunxu , Liu Yang , Xu Lin , Liu Deruo , Hu Jian , Zhao Heng , Sun Wei , Yang Haiying
Published: May 2019

e15519

Background: The Lymph node metastasis (LNM) trends toward inferior outcomes for the esophageal cancer (EC) patients after radical operation. However, it is not well known for status to metastasis of regional lymph node (RLN) and impacts of RLN metastasis on long-term survival following esophagectomy, especially in squamous cell carcinoma. Methods: Data was extracted from the Collaborative Prospective Esophageal Cancer Database (designed by LinkDoc Technology Co, Ltd.) for patients with stage T1-3 thoracic tumors who underwent radical esophagectomy between 1 January 2010 and 30 June 2017 across ten hospitals with high volume of surgeries in China. The rate of LNM in every lymph node station was defined as the number of patients with LNM in the station divided by the number of patients with examination of lymph node (ELN) in the station. The hazard ratios (RLN metastasis versus no RLN metastasis) for postoperative survival were analyzed by method of multivariate cox’s proportional hazard models, and confounders such as age, sex, histological grade, stage T, tumor size and neoadjuvant therapy were adjusted. Results: Total 6485 patients (mean age: 61.80±8.13) were included, composing of 75.2% male and 24.8% female. For tumor types, there were 95.5% squamous cell carcinoma, 2% adenocarcinoma and 2.5% others. The rates of RLN metastasis and hazard ratios are in the table. Conclusions: Besides lymph node station 8, LNM was commonly found at station 1,2,4,16,17 and 20 in thoracic stage T1-3 EC. The RLN metastasis predicted inferior postoperative long-term survival. [Table: see text]
Expression of CD44 and CD133 in tumor сells of metastatic and non-metastatic gastric cancer.
Authors: Sagakyants Aleksandr B. , Kit Oleg I. , Ulianova Elena P. , Zlatnik Elena Yu. , Novikova Inna A. , Shulgina Oksana G. , Gevorkyan Yuriy A. , Soldatkina Natalya V. , Samoylenko Nikolay S. , Vladimirova Liubov Yu
Published: May 2019

e15531

Background: High incidence and mortality rates of gastric cancer cause a constant search for the most informative and effective methods of diagnosis and treatment assessment. In this regard, studying the expression of markers with the stem phenotype in primary tumor tissues in patients with gastric cancer with and without metastases is of undoubted interest. Methods: The study included 20 gastric cancer patients aged 30-80 years: group 1 – gastric cancer T3-4аN0-3M0 (G2) without metastasis (58.9±9.7 years); group 2 – gastric cancer T3-4аN0-3M1 (G2) with peritoneal metastasis (53.4±11.9 years). Immunohistochemical study was performed on paraffin-embedded tumor tissue sections using mouse monoclonal antibodies to CD44 (156-3С11 Thermo Scientific) at a 1:2500 dilution and rabbit polyclonal antibodies to CD133 (Cloud-Clone Corp.) at a 1:700 dilution; the Thermo Scientific autostainer was used for staining. Membrane staining and staining intensity were assessed: 0, 1+ weak, 2+ moderate, 3+ strong staining. Positive expression was defined as ≥10% cut-off for CD44 and ˃5% for CD133. Results: Positive expression of CD44+ was detected in 67% (13) in group 2 vs. 20% (4) in group 1. In the metastatic group, the number of cells that stained positive for CD44 expression ranged from 9 to 15%, on average 10.0±3.08%, without metastases – from single cells to 13%, on average 6.0±2.3%. A chi-square test showed statistically significant association in the groups (8.256 at p = 0.004). Positive CD133+ expression in tumor tissues was registered in 100% (20) in group 2 and 80% (16) in group 1. The range of positively stained cells in group 2 was from 10 to 40%, on average 21.3±11.6%, in group 1 - from single cells to 14%, on average 10.0±2.4%. A chi-square test showed statistically significant association in the groups (4.444 at p = 0.036). Conclusions: Immunohistochemical study of the selected tumor cell markers in gastric cancer revealed some characteristics of their expression depending on the presence of metastases. The results can be the basis for further research for the most complete characterization of a heterogeneous tumor population in gastric cancer and the role of individual cells in the tumor growth, progression and metastasis.
Apatinib inhibits tumor progression and promotes antitumor efficacy of cytotoxic drugs in esophageal squamous cell carcinoma.
Authors: Chi Yanyan , Wang Feng , Meng Xiangrui , Shan Zhengzheng , Sun Yan , Fan Qingxia
Published: May 2019

e15554

Background: Apatinib, a highly selective inhibitor of vascular endothelial growth factor receptor-2 (VEGFR-2), inhibits the angiogenesis of tumors. The function and mechanism of Apatinib in esophageal squamous cell carcinoma (ESCC) remains unknown. Methods: The expression of VEGFR-2 in ESCC cell lines (KYSE450, EC1, KYSE410, KYSE70) were detected by western blot. KYSE450 and EC1 cell lines were treated with Apatinib, or combined with cytotoxic drugs: paclitaxel (TAX), 5-fluorouracil (5-FU) or cisplatin (DDP) respectively. Cell proliferation was then measured using CCK-8 assay; cell apoptosis was analyzed by flow cytometry; cell migration and invasion were evaluated by wound healing and transwell assays. The expression of VEGFR-2, Bcl2, MMP-2/MMP-9, p-Akt and p-mTOR in KYSE450 and EC1 cell lines were determined by western blot. Esophageal cancer xenografts model was established and used to evaluate the antitumor effects of combination of Apatinib and cytotoxic drugs in vivo. Immunohistochemistry was used to detect the expression of Ki67, VEGFR-2 and CD31 in tumor tissues of esophageal cancer xenografts model. Results: We found that Apatinib efficiently inhibited cell growth, metastasis and activity of the Akt/mTOR pathway in ESCC cells. Western blot analysis showed that Apatinib significantly increased Bax protein levels, decreased VEGFR-2, Bcl2, MMP-2/MMP-9, p-Akt and p-mTOR protein levels in ESCC cells. Moreover, Apatinib enhanced chemosensitivity of cytotoxic drugs TAX, 5-FU and DDP by upregulating expression of Bax protein, and downregulating expression of VEGFR-2, Bcl2, MMP-2/MMP-9 protein in vitro. Compared with single agent groups, the combination of Apatinib with each chemotherapeutic drug significantly repressed tumor growth and angiogenesis through blocking the expression of Ki67, VEGFR-2 and CD31 in vivo. Conclusions: Taken together, Apatinib suppressed cell growth, migration and invasion, and promoted antitumor effect of chemotherapeutic agents in ESCC.
Tumor copy number alteration (CNA) burden as a prognostic factor for overall survival in Chinese gastric cancers.
Authors: Han Bo , Ren Dandan , Mao Beibei , Song Xue , Yang Wanning , Zhang Henghui , Gao Feng
Published: May 2019

e15555

Background: Gastric cancer (GC) is associated with high morbidity and mortality rates in the world with poor prognosis and limited treatment options. The level of copy number alteration (CNA), termed CNA burden, is reported as a pan-cancer prognostic factor associated with recurrence and death. The current study aims to investigate association between CNA burden in primary tumor tissue and overall survival (OS) of Chinese patients with GC after surgical resection. Methods: The present study included 78 patients who had received surgical resection and adjuvant chemotherapy with poorly differentiated GC. The primary outcome was OS. Tumor specimens were obtained from surgery and submitted for next generation sequencing (NGS) with matched normal tissue samples. A 1408-gene panel was used to identify genome profiles. Data were analyzed using Cox proportional hazards models and Kaplan-Meier survival analysis. Results: The most frequently altered genes were TP53 (47%), PIK3CA (10%), PTEN (9%), NOTCH1 (8%) and RNF43 (6%), and copy numbers of TRPS1 (65%), COL1A2 (50%), CSMD3 (45%), ZFHX4 (45%), NAV3 (36%) varied most frequency in current cohort. Greater tumor CNA burden correlated with an increase in death from disease compared to a lower tumor CNA burden ( p= 0.0066). In addition, there were statistically significant differences in OS between different clinical staging ( p= 0.0011). Moreover, the Cox proportional hazard model showed that CNA burden was an independent prognosis factor in GC. Finally, we performed an independent signature that includes CNA burden and clinical staging to predict survival of GC. Conclusions: This study indicates that tumor CNA burden is an independent predictive survival biomarker for Chinese gastric cancers. CNA burden combined with clinical staging is a better predictor for postoperative survival prediction of gastric cancer.
Comparison of tumor characteristics and survival between esophageal and other gastroenteropancreatic neuroendocrine neoplasms.
Authors: Taherian Mehran , Samankan Shabnam , Groman Adrienne , Yendamuri Saikrishna S. , Bhalla Amarpreet
Published: May 2019

e15556

Background: Neuroendocrine neoplasms (NEN) of esophagus are extremely rare. Limited information is available on survival of these tumors. The objective of this study was to define the clinicopathologic predictors of overall survival (OS) in esophageal NEN, and to compare them with the other gastroenteropancreatic NEN (GEP-NEN). Methods: Esophageal NEN were selected from the National Cancer Database (2004–2013). Multivariable analysis and Kaplan–Meier method were performed. The prognostic factors for GEP-NEN were derived from literature including WHO classification and AJCC TNM classification. Results: Of 802 selected patients with esophageal NEN, 97.5% were NEC and only 2.5% typical NET. The median age for NET was 58 vs. 66 for NEC (p = 0.007). NET more commonly presented in females (60%) compared to NEC wherein 68% patients were male. Most of the NEC were grade III/IV and > 4 cm, while most NET were grade I/II and < 4 cm. They most frequently metastasized to the liver. 10.7% of patients with esophageal NEN underwent esophagectomy while 86.5% had no surgery; 68.5% had adjuvant and 6.6% neoadjuvant therapy. Multivariable analysis showed that tumor > 4 cm (hazard ratio (HR) 1.45; P = 0.013), stage III and IV (HR 2.27; p = 0.030, and HR 4.02; P < 0.001, respectively) were associated with significantly worse OS, while esophagectomy (HR 0.30; P = 0.019) and neoadjuvant therapy (HR 0.35; p = 0.006) were predictors of better OS. The 5-year OS rate was 12% for all esophageal NEN (95% CI, 10-15): 89% for NET and 9% for NEC. Pancreatic NET are generally > 2 cm and NEC have an average size of 4 cm. The factors associated with worse prognosis in pancreatic NEN include positive surgical resection margins, lymph node metastases, advanced TNM stage, vascular invasion and distant metastasis. The 5-year OS for patients with pancreatic NET and NEC is 65% and 16%, respectively. NET of ileum are < 2 cm in 47% of cases, and the 5-year OS is about 60%. Tumor stage is the most important predictor of survival. The malignant potential is retained for ileal NET > 1 cm. Only Stage III vs. Stage IV has a better OS. The G3 NEC and mixed neuroendocrine-non-neuroendocrine neoplasms (MiNEN) have poor OS and variable median survival time reported up to 40 months. Conclusions: Using the largest dataset of esophageal NEN to date, the major independent predictors of OS include tumor size, stage, esophagectomy, and chemotherapy. The majority of esophageal NEN are NEC. Their OS is similar or slightly higher in comparison to ileal NEC but poor in comparison to similar tumors presenting in the pancreas.
The genetic profiles of esophageal squamous cell carcinoma (ESCC) in Hebei, a northern province of China.
Authors: Zuo Jing , Jia Yimeng , Wang Yudong , Fan Zhisong , Wang Long , Su Yu , Liu Fengling , Jiang Da
Published: May 2019

e15558

Background: Esophageal squamous cell carcinoma (ESCC) is a common malignancy without effective therapy. Methods: DNA was extracted from formalin-fixed, paraffin-embedded (FFPE) sections from 29 ESCC samples. A panel of 520 genes was sequenced with NGS to identify genomic alterations at an average sequencing depth of 1000×. Results: In total, 421 genomic alterations were identified. We identified 230 single nucleotide variants (SNVs) and 35 insertions and deletions (indels) , 147 CNAs, 9 deletions and 0 translocations. The most frequently mutated genes were TP53 (97%), NOTCH1 (31%), CDKN2A (21%), NFKBIA (21%). Most frequent CN gains were observed in FGF3/4/19/CCND1(41%) which were co-occurrence in cBioPortal.By performing targeted NGS in 29 ESCC samples, we found a median TMB of 5.6/Mb (range: 0.8 to 42.9/Mb).TMB-high was located according to TMB > = 10, where 17.2% of the samples were TMB-H.C·G > T·A transversions were the most common mutations, followed by G·C > A·T transitions,The patients with NFKBIA mutation were all significantly older (p = 0.05), and the patients with FGF3/4/19/CCND1 amplification were mostly moderately differentiated, and the patients without high differentiation, suggesting a higher degree of malignancy (p = 0.03). CDKN2A, FGF3/4/19/CCND1 amplification, and NFKBIA mutated were associated with a smaller invasion depth, respectively (p = 0,p = 0.03,p = 0.05). Patients with wild type KMT2D had earlier stage, and all N0 stages were wild type for KMT2D (p = 0.03).Notably, high activity of DNA damage,Cell cycle and Notch signaling pathway might be particularly promising strategies for ESCC.In potential therapeutic target analysis, CDKN2A(20.7%) and PIK3CA (13.8%) were the validated and candidate therapeutic targeted genes. Conclusions: Collectively, our data provide comprehensive insights into the mutational signatures of ESCC and identify markers for early diagnosis and potential therapeutic targets.
Management of synchronous Krukenberg tumors from gastric cancer: A single-center experience.
Authors: Yu Pengfei , Huang Ling , Du Yian , Cheng Xiangdong
Published: May 2019

e15563

Background: Whether metastasectomy improves prognosis of gastric cancer patients with ovarian metastases (Krukenberg tumors) is not clear. In this study, we examined the survival benefit of metastasectomy combined with chemotherapy for treatment of synchronous Krukenberg tumors from gastric cancer and identified the prognostic factors. Methods: The subjects of this study were patients diagnosed as synchronous Krukenberg tumors of gastric origin in the period between December 2004 and December 2015. Patients were classified in accordance with treatment modality: metastasectomy group (metastasectomy combined with chemotherapy) and non-metastasectomy group (chemotherapy alone). Clinicopathological characteristics together with treatment records were investigated in detail and their relationship with survival outcomes was examined. Results: Out of a total of 103 patients, 54 (52.4%) underwent metastasectomy of Krukenberg tumors while 49 (47.6%) patients had chemotherapy alone. Overall survival (OS) in the metastasectomy group was significantly longer than that in the non-metastasectomy group (18.9 months vs. 12.4 months, respectively; P < 0.001). Metastasectomy (hazard ratio [HR] 0.486; 95% confidence interval [CI] 0.323–0.729; P < 0.001), signet ring cells (HR 1.938; 95% CI 1.182–3.175; P = 0.009), peritoneal carcinomatosis (HR 1.934; 95% CI 1.230–3.049; P = 0.004), expression of estrogen receptor-β (ER-β) (HR 0.404; 95% CI 0.251–0.648; P < 0.001), and progesterone receptor (PR) (HR 0.496; 95% CI 0.301–0.817; P < 0.001) were independent predictors of OS. Conclusions: Metastasectomy combined with chemotherapy showed an association with survival benefit in patients with synchronous Krukenberg tumors from gastric cancer. Metastasectomy, expression of ER-β and PR, peritoneal carcinomatosis, and signet ring cells were independent predictors of survival. Further prospective studies are warranted.
Integrating endogenous peptides analysis and protease mapping for identification of potential serum biomarkers in gastric adenocarcinoma.
Authors: Forones Nora Manoukian , Oliveira Talita Mendes de , Lacerda Jose Thales , Dias Meiriellen , Mendes Maria Anita , Juliano Maria Aparecida
Published: May 2019

e15564

Background: Gastric cancer (GC) is the third cause of cancer death in the world. In Brazil 21.000 new cases had been diagnosed in 2018 and the mortality rate is associated with late diagnosis that contributed to poor prognosis. The objective was to study endogenous peptides in serum of patients with GC. Methods: Fifteen serum samples of patients with diagnosis of gastric adenocarcinoma (TNM stage I-IV) and 15 controls were included. Endogenous peptides were extracted and pooled of random mode in 5 biological replicates (n = 3 samples) for each group. The mixture peptides was submitted to nLC-MS/MS analysis and peptide sequences identified were used to protease mapping. Results: A total of 191 peptides (≥ 7 amino acids) were identified corresponding to 36 proteins involved mainly on metabolic and immune system processes, signal transduction, platelet and neutrophils degranulation. Peptidome-based protease mapping identified 59 proteases (29 serine-, 19 metallo-, 8 cisteine- and 3 aspartic-), in which Prothrombin, Plasminogen, MMP14, MMP7 and MMP3 proteases yielded most of peptides. Twenty sequences derived from Fibrinogen A, Fibrinogen B, Complement C3 (C3f), Apolipoprotein A-I (N-terminal), Prothrombin (C-terminal of Activation peptide fragment 2; N-terminal of Thrombin light chain) and Coagulation factor XIII A (N-terminal) were significantly different (p ≤ 0.5). Between these peptides, 14 were up regulated and six downregulated in GC patients. The phosphopeptide 20ADSpGEGDFLAEGGGVR35 (Fibrinopeptide A) was significantly increased (p< 0.03) in GC serum samples, while the non-phosphorylated Fibrinopeptide A was decreased (p<0.04), suggesting a higher phosphorylation rate in this protein in GC patients. Further, the peptide 31QGVNDNEEGFFSAR44 (Fibrinopeptide B) was downregulated (p<0.04) in GC serum samples; in counterpart, the Fibrinopeptide B derived peptides NDNEEGFF (p<0.0007) and QGVNDNEEGFFS (p<0.0013) was up regulated, suggesting a higher proteolysis in GC serum samples. Conclusions: The investigation of protease/substrate activity relationship may be described such as a novel panorama for discovery of serum biomarkers.