bluuebunny/crossref_metadata_2025 huggingface dataset:
PD-L1 and PD-1 expression profile depending on the microsatellite status and the histological subtype in colorectal carcinomas.
Authors: Bossard CÃ©line , Ott Eva , Dansette Delphine , Ouairy Adrien , Jarry Anne , Bezieau Stephane , Toquet Claire , Mosnier Jean-François
Published: March 2017

590

Background: PD-1/PD-L1 blockade showed therapeutic efficacy in only microsatellite (MSI) colorectal carcinomas (CRC), however, the profile of PD-L1 and PD-1 expression in CRC is only partially described. Methods: We thus analyzed on FFPE whole-tissue sections of 80 CRC, the expression profile of PD-L1 by tumor and/or immune cells by immunohistochemistry (clone E1L3N) depending on the MSI status and the histological subtype, and correlated to the density of PD-1+ and Tbet+ (able to secrete IFNg known to induce PD-L1) tumor-infiltrating lymphocytes (TIL). Results: 78% of MSI CRC (32/41) overexpressed PD-L1 either by tumor or immune cells versus 46% of MSS CRC (18/39) (p 0.005). This overexpression was heterogeneous within the same tumor in most of cases. Among MSI CRC, PD-L1 was preferentially overexpressed in medullary carcinomas (MC, 19/21, 90%) compared with 65% (13/20) in non-medullary adenocarcinomas (p 0.06). PD-L1 expression by tumor cells was only observed in MSI CRC (19/41, 46% with PD-L1 expression in more than 5% of tumor cells – score 1), and preferentially in MC (57% vs 5% in no medullary adenocarcinomas, with PD-L1 expression in more than 50% of tumor cells – score 3, p 0.0005). Conversely, PD-L1 expression by immune cells was observed in MSI CRC (23/41, 56% with PD-L1 expression by more than 5 sheets of 50 positive cells) but also in MSS CRC (18/39, 43%) (p 0.5). The density of PD-1+ cells was significantly correlated to the PD-L1 expression, as well as the density of Tbet+ TIL. Conclusions: PD-L1 expression is 1) heterogeneous in CRC, among CRC but also within the same tumor, 2) preferentially observed in MSI CRC (78%), especially in MC (90%), where PD-L1 is expressed by tumor cells, 3) correlated with the density of PD-1+ or T-bet+ TIL, and 4) observed in a significant proportion of MSS CRC (46%) by immune cells only. From a clinical point of view, PD-L1 expression has to be determined at best in full tissue section and besides its preferential expression in MSI CRC, its significant frequency and expression profile (only by immune cells) in MSS CRC should be taken into account in the future clinical trials testing the efficacy of anti-PD-1/PD-L1 antibodies.
Placental growth factor and the angiogenic environment based on analysis of baseline plasma biomarkers from the VELOUR trial.
Authors: Tabernero Josep , Paccard Caroline , Chiron Marielle , Dochy Emmanuelle , Van Cutsem Eric
Published: March 2017

592

Background: A number of available agents (e.g., bevacizumab [bev], ziv-aflibercept, ramucirumab, regorafenib) for metastatic colorectal cancer (mCRC) target the VEGF pathway; biomarkers for anti-VEGF response are thus valuable. The Ph III VELOUR study (NCT00561470) demonstrated that addition of aflibercept to FOLFIRI improved survival and response rates in mCRC patients (pts) who had failed oxaliplatin-based therapy, including pts treated with prior bev. We report the correlation between biomarkers and bev pre-treatment in baseline plasma samples from VELOUR pts. Methods: 98 biomarkers were examined in baseline plasma samples from pts enrolled in VELOUR. Biomarker concentration was determined using multiplex assays or ELISA. Results: Plasma samples were available for 553 pts (placebo n = 265; aflibercept n = 288), of whom 169 had received prior bev. Nine biomarkers implicated in angiogenesis or bev resistance correlated with prior bev therapy (Table), including VEGF-A (~5-fold higher concentration) and PlGF (~2-fold higher concentration). Conclusions: These data indicate that prior bev therapy is likely to modulate expression of biomarkers associated with acquired resistance to anti-VEGF treatment (e.g., VEGF-A & PlGF). However, the improvement in clinical outcomes with aflibercept + FOLFIRI (vs placebo + FOLFIRI) in bev pretreated pts in VELOUR suggests that aflibercept remains active despite the development of bev-induced angiogenic resistance. Clinical trial information: NCT00561470. [Table: see text]
Is the primary tumor location (PTL) associated with differential gene expression profiles in patients with metastatic colorectal cancer (mCRC)? Analysis of the FIRE1-trial.
Authors: Le Lien D. , Mansmann Ulrich Robert , Jung Andreas , Kirchner Thomas , Schäfer Reinhold , Neureiter Daniel , Holch Julian Walter , Heinemann Volker , Stintzing Sebastian
Published: March 2017

598

Background: Site-differential molecular characteristics of PTL as well as its influence on overall survival have been reported. In our study, we investigated the association of PTL with gene- and pathway-wise RNA expression as well as its representation in RAS mutational status subgroups. Methods: The phase-III-trial FIRE-1 compared the efficacy of FuFIRI and mIROX in patients with mCRC. Formalin-fixed, paraffin-embedded pretreatment samples from 166 FIRE-I patients were collected. The splenic flexure was used for differentiation between left colon cancer (LC) and right colon cancer (RC). RAS mutated PTs (RASmut) had PCR confirmed mutations in KRAS gene (Exon 2, 3, or 4) or in NRAS gene (Exon 2, 3, or 4). RNA expression profiling used the nCounter(R) PanCancer Pathways Panel including DNA damage control and TGF-β. We also studied a set of selected genes (Aprataxin, AREG, CD3EAP, CES1, CES2, CMPK1, DPD, DUT, ERCC1, ERCC5, EREG, FGF2, GSTP1, KLC3, MTHFR, PPP1R13L, RCC1, TOPO1, TP, TS, and UGT1A1). The gene-wise analyses used Limma and the permutated t-test with appropriate multiple testing adjustment. Holistic pathway investigation used GlobalAncova and the global tests. RAS mutation status and its correlation with tumor side were included as factors in both analyses. Additionally, gene- and pathway-wise analyses were performed in RASmut and RASWTsubgroups. Results: A total of 83 (50%) of 166 patients had any RAS mutation. A total of 28 (17%) patients had PTs on the right side and 13 of those (46%) were RASmut. The top ten differentially expressed genes between LC and RC are MMP9, ERCC3, MAP3K1, RRAS2, SOCS3, RB1, TGFβ1, LEFTY1, NKD1, and CDC25B (unadjusted p-values < 0.005). Potential enrichment in the DNA damage control pathway was found (p = 0.008). The RAS subgroup analyses showed a non-overlapping top ten list. Side specific effects may exist for the TGF-β pathway and the selected gene set in RASWTpatients. Conclusions: No strong association between PTL and gene expression showed up in mCRC tumors nor RAS subgroups. Potentially differential gene expression may exist for specific genes, two pathways and our selected gene set.
A prospective multicenter observational study of surgical palliation examining postoperative quality of life in patients treated for malignant gastric outlet obstruction caused by incurable advanced gastric cancer.
Authors: Fujitani Kazumasa , Ando Masahiko , Sakamaki Kentaro , Terashima Masanori , Kawabata Ryohei , Ito Yuichi , Yoshikawa Takaki , Kondo Masato , Kodera Yasuhiro , Kaji Masahide , Oka Yoshio , Imamura Hiroshi , Kawada Junji , Takagane Akinori , Shimada Hideaki , Tanizawa Yutaka , Yamanaka Takeharu , Morita Satoshi , Ninomiya Motoki , Yoshida Kazuhiro
Published: March 2017

6

Background: Decision-making for surgical palliation remains one of the most challenging clinical scenarios since quality of life (QOL) is a key component of cancer care. We conducted this study to examine the impacts of surgical palliation on postoperative QOL in patients (pts) with malignant gastric outlet obstruction (GOO) caused by incurable primary gastric cancer (GC). Methods: Eligibility included (1) no oral intake or liquids only requiring parenteral nutrition (2) aged ≥20 (3) surgically fit (4) ECOG PS of 0-2 and (5) written IC. Patients underwent either palliative distal/total gastrectomy (DG/TG) or gastrojejunostomy (GJS). Treatment choice was left to the discretion of the physician. Validated QOL instruments (EORTC QLQ-STO22 and EuroQol-5D) assessed QOL at baseline, 2 weeks (wks), 1 month (m), and 3 months following the surgical palliation, and two observational outcomes (postoperative improvement of oral intake, and safety of surgical intervention) were evaluated. Results: 104 pts, 71 males and 33 females with a median age of 68 years, were enrolled. The types of surgery were DG in 23 pts, TG in 9 pts, GJS in 70 in pts, and exploratory laparotomy in 2 pts. Baseline QOL questionnaires were completed by 103 (99.0%) pts. Among the 104 pts, 98 (94.2%), 100 (96.1%), and 81 (77.9%) completed the 2-wk, 1-m, and 3-m follow-up survey, respectively. The mean baseline EQ-5D score was 0.74 (SD, 0.21). During the follow-up period, the mean scores remained consistent with the baseline scores; the change from baseline score was within ± 0.05 for the index. Many pts came to eat solid food at 2 wks postsurgery and remained tolerable thereafter (from 0 at baseline to 82, 85, 75 pts at 2 wks, 1 m, and 3 ms, respectively). Overall morbidity rate of ≥grade 3 on Clavien-Dindo classification and 30-day postoperative mortality rate was 9.6% (10 pts) and 2.0% (2 pts) with a median hospital stay of 13 days and re-operation rate of 3.9% (4 pts). Conclusions: In pts with malignant GOO caused by advanced GC, surgical palliation maintained patient QOL while improving solid food intake with an acceptable surgical safety. Clinical trial information: UMIN000023494.
Effect of polymorphisms of genes encoding regulatory proteins in the coagulation cascade on outcome for mCRC patients treated with FOLFIRI and bevacizumab: Data from FIRE-3 trial.
Authors: Berger Martin D. , Stintzing Sebastian , Yang Dongyun , Suenaga Mitsukuni , Miyamoto Yuji , Cao Shu , Soni Shivani , Gopez Roel , Hanna Diana L. , Zhang Wu , Heinemann Volker , Lenz Heinz-Josef
Published: March 2017

601

Background: Components of the coagulation cascade and surface proteins of platelets are involved in the regulation of angiogenesis. There are preliminary data suggesting that antithrombin III (ATIII), beside its anti-angiogenic effects, might also inhibit cancer cell migration and invasion. We therefore hypothesize that genes encoding regulatory proteins within the coagulation cascade and surface proteins of platelets may predict outcome in patients with metastatic colorectal cancer (mCRC). Methods: 295patients with mCRC enrolled in the randomized phase III FIRE-3 trial and treated with FOLFIRI and bevacizumab were included in this study. Genomic DNA was extracted from formalin fixed paraffin embedded tissue. 6 functional SNPs in 6 genes (FGB, F2, F2R, SERPINC1, ITGA2B, ITGB3) were analyzed by PCR-based direct sequencing. Candidate SNPs and their correlation with outcome were analyzed by uni- and multivariate analysis. Results: Baseline characteristics were as follows: female/male = 99/196; median age = 65y (31-76) and median PFS/OS = 10.1/24.2 months. The SERPINC1 variant rs5877 encoding ATIII and the FGB SNP rs4220 encoding fibrinogen showed significant association with overall survival (OS). Patients with a C/C genotype of the SERPINC1 SNP rs5877 had a significantly worse OS than those harboring any T alleles (17.6 vs 25 months) in both univariate (HR 1.66, 95% CI 1.11-2.48, p = 0.011) and multivariate analysis (HR 1.74, 95% CI 1.15-2.64, p = 0.009). Similarly, A/A genotype carriers of the FGB SNP rs4220 showed a shorter OS compared to those with any G alleles (14.7 vs 24.8 months) in univariate (HR 1.90, 95% CI 1.05-3.42, p = 0.028) and multivariate analysis (HR 2.83, 95% CI 1.49-5.40, p = 0.002). Conclusions: This is the first report demonstrating that the SERPINC1 polymorphism rs5877 and the FGB SNP rs4220 might serve as predictive/prognostic biomarkers in patients with mCRC treated with FOLFIRI and bevacizumab in the first line setting.
Clinical applications of extended ctDNA RAS mutation determination in metastatic colorectal cancer.
Authors: Vidal Barrull Joana , Muinelo Laura , Dalmases Alba , Abalo Alicia , Vela Maria Carmen , Brozos EM , Muset Maria Merce , Ruiz BaÃ±obre Juan , Iglesias Mar , Blanco Cristina , Lopez Eva , Rodriguez Lopez Carmela , Jones Frederick S. , Edelstein Dan , Lukas Antje , Albanell Joan , Bellosillo Beatriz , Candamio Sonia , Montagut Clara , Lopez Rafael
Published: March 2017

607

Background: Tumor tissue is currently used for RAS testing in metastatic colorectal cancer (mCRC) patients, but detection of circulating tumor (ct) DNA in plasma is being actively investigated as an alternative for detection and monitoring RAS mutations during therapy. Methods: Concordance of RAS testing in plasma using the OncoBEAM RAS CRC test and RAS testing in tissue using standard-of-care RAS detection techniques was evaluated in 114 mCRC antiEGFR-naïve patients. In discordant cases, tissue samples were re-examined by BEAMing. OncoBEAM RAS CRC assay was also used to monitor RAS mutation status in serial plasma samples from 34 patients treated with anti-VEGF +/- chemotherapy or anti-EGFR based therapy. Results: The overall agreement of the two methods was 93% (106/114 patients), with positive agreement of 96.2% (51/53) and negative agreement of 90.2% (55/61). Longitudinal monitoring of plasma samples from 13 patients with RAS mutated tumors treated with chemotherapy +/- anti-VEGF, showed a significant decrease (or disappearance) of RAS mutant allele fraction (MAF) in all cases at the time of CT-scan (8-12 weeks after onset of treatment). In 3 patients, plasma was also analyzed at week 4 showing the same dramatic decrease in MAF. RAS decline mirrored clinical and radiological response to treatment. Interestingly, RAS decline was also observed in patients with stable disease in CT-scan by RECIST1.1. In 7 of 9 patients that subsequently progressed, RAS ctDNA fraction increased accordingly. On the other hand, among 22 patients RAS wt treated with cetuximab, 6 showed emergence of RAS mutations at progression. In two cases, multiple RAS mutations were concomitantly present. Conclusions: The high overall agreement between basal plasma and tissue RAS mutation status indicates that blood-based testing with OncoBEAM RAS CRC is a viable alternative to tissue RAS testing in mCRC patients. Moreover, this study highlights the utility of OncoBEAM RAS CRC to monitor treatment of mCRC patients in two setting: (a) evaluation of response to anti-VEGF +/- chemotherapy in RAS mutant patients, as a complement to RECIST radiological criteria and (b) to monitor emergence of resistant clones during anti-EGFR treatment in RAS wt patients.
Association of chromosome 17 copy number instability with favorable prognosis in nonsurgically treated gastroesophageal adenocarinoma and impaired response to trastuzumab.
Authors: Spada Neal , Birkness Jacqueline , Dacic Sanja , Luketich James D. , Nason Katie Sue , Sun Weijing , Davison Jon M.
Published: March 2017

61

Background: Chromosomal instability (CIN), defined as cell to cell variation in copy number, is a feature of solid tumors that has been shown in experimental systems to result in aneuploidy, DNA damage, changes in gene expression, and genetically heterogeneous cell populations. In esophageal adenocarcinoma, chromosome 17 (Chr17) aneusomy is associated with heterogeneous HER2 amplification, a marker of poor prognosis (Yoon et al., J Clin Oncol 2012), but the association between CIN, HER2 amplification and clinical outcome has not been well studied. Methods: We retrospectively analyzed individual cell Chr17 centromere counts in 348 gastroesophageal adenocarcinomas that were tested for HER2 amplification. As an estimate of Chr17 CIN (CIN-17), we calculated the percentage of cells with centromere counts differing from the mode (modal centromere deviation, MCD, Roylance et al. Cancer Epidemiol Biomarkers Prev, 2011). We analyzed the association of CIN-17 with Chr17 aneusomy, HER2 amplification, pathologic tumor characteristics and clinical outcome. Results: Using pre-established cutoffs, we found CIN-17 (MCD > 30%) in 45% (158/348) and extreme CIN-17 (MCD > 45%) in 28% (99/348) of cases. Compared to CIN-17 negative tumors, CIN-17 positive tumors were more likely to be polysomic (77% vs 0.5%, P < 0.001) and Lauren intestinal type (84% vs 70%, P = 0.005). HER2 amplification was detected in 23% (80/348) of tumors, but there was no association with CIN-17 (P = 0.493). In patients who received pharmacologic therapy (n = 77), there was a 58% reduction in overall mortality associated with extreme CIN-17 (HR = 0.42, 0.25-0.72) independent of age, stage and addition of trastuzumab. Addition of trastuzumab to pharmacologic therapy showed a trend toward improved clinical outcome only in the subgroup without extreme CIN (n = 50, HR = 0.37, 0.13-1.06). CIN-17 was not associated with differences in patient survival after surgical resection. Conclusions: In this retrospective study, extreme CIN-17 was a favorable prognostic factor in patients who receive chemotherapy but could impair response to trastuzumab. These findings warrant further study.
Prognostic value of PD-L1 expression in colorectal cancer.
Authors: Lin Albert Y. , Kouzminova Natalia B. , Chou Thomas
Published: March 2017

610

Background: Growing evidence have shown promise for targeting programmed death-1 (PD-1)/programmed death-ligand 1 (PD-L1) signaling in several tumors. However, the role of PD-L1 expression in colorectal cancer (CRC) tumor cells and its interaction with other clinicopathologic factors remain elusive. Methods: We constructed a tissue microarray with paraffin-embedded primary colon cancer tissue (n=103; stage I, 22; II: 44; III: 37). Expression levels of biomarkers (PD-L1, EGFR, p53, Ki67, LEF1, VEGF, COX2, MMR – by IHC, and MiR-34a - by in situ hybridization), clinicopathologic variables and clinical outcomes were investigated. H-score evaluation of PD-L1 was performed. Tumors with an H score >55, derived using Cutoff Finder, were considered PD-L1 positive for use in Kaplan-Meier survival analysis with log-rank test and in Cox regression models for recurrence-free survival (RFS) and overall survival (OS). Results: Median follow-up time for the entire cohort was 4.7 years (range 0.1-20.8). Positive PLD1 was identified in 89% of cases. As a group, cases with positive PD-L1 expression had better OS than those with negative expressions (12.2 vs 3.3 years) (p = 0.043), especially in cases with mismatch repair proficiency (MMR-P, 12.1 vs 1.0 years) (p <0.001). Negative PD-L1 expression in cases who had no chemotherapy was associated with worse outcome for OS (p=0.001) and RFS (p=0.008). There was no significant association between PD-L1 expression and gender, age, tumor size, pathologic stage or tumor grade. However, PD-L1 expression was significantly associated with MiR-34a (p<0.001) and p53 overexpression (p=0.008) with a trend toward association in cases with mismatch repair proficiency (MMR-P, p=0.053). Conclusions: Our results suggest that negative PD-L1 expression appears to have negative prognostic value with worse RFS and OS in stage I - III CRC patients, especially those who had not received any chemotherapy. Given a recent study suggesting p53/miR-34a/PD-L1 axis as a novel mechanism of immune evasion, and strong correlation in the PD-L1/p53 and PD-L1/miR-334a expressions in our cohort, further investigation of their interactions is warranted that may lead to identifying their predictive value for immuno/chemotherapy.
Magnetic resonance Imaging (MRI), liquid biopsies, and patient derived organoids (PDOs) as biomarkers of response to regorafenib (REG) in treatment-refractory metastatic colorectal cancer (mCRC) patients (pts).
Authors: Khan Khurum Hayat , Rata Mihaela , Koh Dow-Mu , Tunariu Nina , Vlachogiannis George , Hahne Jens , Jamin Yann , Collins David J. , Chau Ian , Rao Sheela , Watkins David J. , Starling Naureen , Kalaitzaki Eleftheria , Fotiadis Nicos , Begum Ruwaida , Temple Eleanor , Fassan Matteo , Braconi Chiara , Cunningham David , Valeri Nicola
Published: March 2017

613

Background: REG demonstrated efficacy in pre-treated mCRC pts. Lack of predictive biomarkers, potential toxicities and cost/effectiveness concerns highlight the unmet need for better patient selection. Methods: RAS mutant mCRC pts with biopsiable metastases were enrolled in this phase II trial. Tissue biopsies (6-12 cores) were obtained at baseline (BL), after 2 months if stable disease (SD) and at disease progression (PD). Dynamic contrast enhanced (DCE) MRI was acquired pre and at day 15 post-treatment. Median values of volume transfer constant (Ktrans) and enhancing fraction (EF) [K-trans*EF/100] were generated. Circulating tumour (ct)DNA was collected monthly until PD and tested for clonal RAS mutations by digital droplet PCR. PDOs derived from responders and non-responders pts were implanted orthotopically in the liver of mice and treated with REG for 5 days. Changes in tumour and fractional blood volume (fBV) were monitored by oxygen-enhanced MRI. Results: mCRC pts (n = 27) with paired MRI scans were analysed; a single target lesion per pt was chosen (25 liver and 2 pelvic metastases). Median K-trans*EF/100 product decrease was 58.2%. In the 23 analysable pts (4 received < 1 cycle of treatment due to toxicities), > 70% drop in K-trans*EF/100 (8/23) was associated with higher disease control rate (6/6 vs. 0/6, p = 0.048) measured by RECIST 1.1 at 2 months, improved progression free survival (PFS) [HR = 0.24 (0.07-0.86), p = 0.03], and 4-month PFS (58.3% VS 21.2%). Sequential tissue biopsies analysis confirmed reduction in CD31 in pts with K-trans*EF/100 drop. RAS mutant clones decay in ctDNA after 8 weeks of treatment was associated with better PFS [HR = 0.25 (0.08 - 0.83), p = 0.02] independently of K-trans*EF/100 drop. PDOs xeno-transplants treated with REG compared to controls had significant lower tumour fBV (4.5 VS 10.6, p = 0.03) and lower microvascular density measured by CD31 staining (4.3 VS 8.9, p = 0.02). Conclusions: Combining DCE MRI and ctDNA predicts depth and duration of anti-angiogenic response to REG monotherapy and may improve pt selection with potential health/economic implications. Clinical trial information: 201400357951.
Gaining biomarker insights from existing stage II colorectal cancer (CRC) patient cohorts in the United Kingdom: Using real-world data to guide treatment decisions.
Authors: Fleming Sarah Jane , Morris Eva , Shires Mike , Hemmings Gemma , Wang Lisa , Muranyi Andrea , Singh Shalini , McCusker Margaret Elizabeth , Shanmugam Kandavel , Quirke Philip
Published: March 2017

614

Background: Trial data, while valuable, does not reflect the importance of a biomarker or treatment in the general population as trials generally exclude patients (pts) due to age or comorbidities. We used merged large population datasets to validate a linked anonymized stage II CRC pt cohort as representative of the population to identify the prognostic and predictive value of deficient mismatch repair (MMR) and a new biomarker Ga-interacting vesicle-associated protein (GIV). Methods: English national data on stage II CRC pts surgically treated from 2001-2015, n = 92,147, were analyzed for survival and clinicopathological parameters. Anonymized data were then linked to pathology and a subset of 405 unselected pts surgically treated from 1990-2003 at the Leeds Teaching Hospitals NHS Trust. These were investigated for 5 antibodies. 4 identified deficient MMR status and one was a new marker; GIV. Results: Population data vs the cohort of 405 pts showed a median age of 73 vs 74 yrs, M:F 55%:45% vs. 53%:47%. These are older than seen in most clinical trials and more reflective of the general stage II CRC population. The median survivals of 108 vs 92 months are as expected based on relative time periods covered. 374 patients yielded valid MMR status on immunohistochemistry (92%); 17% were dMMR and 83% were proficient. dMMR vs pMMR pts did not differ in age (median age 75 vs 74.5) or distribution of T3 (69% vs 66%), but were more likely to be female (71% vs 42%), sited in the right colon (76% vs 30%) and poorly differentiated (42% vs 8%). 11% of dMMR and 8% of pMMR received adjuvant chemotherapy in this cohort. GIV scoring status was also evaluated (405): 30% (121) were negative, 65% (264) positive and 5% (20) were not scored. Survival and risk scores by MMR and GIV status will be presented. Conclusions: Population based data has been created to validate the representativeness of a stage II biomarker cohort. This approach using large, population based data-sets provides opportunities to understand the generalizability of biomarker cohorts. The creation and expansion of such cohorts will more effectively validate existing and new biomarkers and treatments in real-world populations.
Tumor heterogeneity in anal cancer assessed with diffusion-weighted (DW) and T2-weighted MRI: Potential biomarker of treatment outcome.
Authors: Owczarczyk Kasia M. , Prezzi Davide , Siddique Muhammad , Stirling James , Cook Gary J. R. , Glynne-Jones Robert , Gaya Andrew M. , Leslie Martin , Goh Vicky
Published: March 2017

617

Background: Despite advances in the management of anal squamous cell carcinoma (ASCC), roughly 25% of patients undergoing chemoradiotherapy (CRT) will experience a disease recurrence. Currently, there is no validated way to predict treatment failure and guidance on the timing of follow-up scans differs. This study was carried out to assess the role of statistical and fractal heterogeneity parameters from pre-treatment and early post-CRT T2- and diffusion-weighted MR in predicting disease outcome. Methods: Baseline and follow-up MR scans from patients with histologically proven non-metastatic ASCC undergoing radical CRT were included in this study. T2-W and DWI MR images were analyzed by two readers, blinded to clinical data. Statistical and fractal heterogeneity parameters were derived using in-house software. Results: 40 patients were included in the final analysis. Follow-up MRI scan was performed at a median of 15 weeks from the start of radiotherapy. 31/40 patients achieved a radiological complete response (CR) and a further 7/40 partial response (PR). Median follow-up was 34.5 months. During the course of follow-up, 12 patients experienced disease recurrence (30%). Radiological CR was not significantly associated with disease outcome. First order statistical heterogeneity analysis demonstrated a significantly lower ROI Skewness (median -0.05±0.49 vs 0.45±1.04 p=0.01) on baseline T2-W imaging as well as lower ROI Entropy (3.95±0.46 vs 4.45±0.49, p=0.01) and higher ROI Energy (0.07±0.03 vs 0.03±0.03, p=0.01) on follow-up T2-W imaging in subjects who experienced disease recurrence. In these patients T2-W Homogeneity derived from Grey-Level Co-occurrence Matrices (GLCM) was lower at baseline (0.41±0.12 vs 0.58 ±0.15 p=0.02) and associated with a higher T2-W Sum Average (median 35±7 vs median 29±7 p=0.01), whereas T2-W Contrast and ADC Correlation derived from GLCM were lower on follow-up imaging (30±11 vs 38 ±35, p=0.01 and 0.77±0.12 vs 0.88±0.12 p=0.04, respectively). Conclusions: First order statistical and fractal heterogeneity analysis of baseline and early post CRT MR scans in patients with ASCC might have a role in predicting long-term disease outcome.
National trends in local excision and esophagectomy for cT1N0 esophageal cancer.
Authors: Sturm Emily C. , Zahnd Whitney , Mellinger John D. , Ganai Sabha
Published: March 2017

62

Background: Esophageal cancer management has evolved due to improvements in staging and treatment strategies. Endoscopic local excision presents an attractive option for definitive management of T1 cancers, avoiding the morbidity of esophagectomy. We hypothesized that for cT1N0 cancers, patients who underwent local excision would have lower survival compared to esophagectomy due to potential discordant staging. Methods: The National Cancer Database was queried for esophageal squamous cell carcinoma (SCC) and adenocarcinoma (AC) with AJCC T1N0 clinical stage who underwent local excision (n = 1625) or esophagectomy (n = 3255) between 1998 and 2012. Chi-square analysis was used to compare demographic and clinical characteristics by procedure. Chi-square trend analysis was performed to assess trends in procedure type over time. Cox Regression analysis was performed to assess survival by procedure controlling for demographic and clinical characteristics. Results: Between 1998 and 2012, the proportion of patients who underwent local excision increased from 12% to 50% for all patients (p < 0.001); from 17% to 40% for SCC patients (p < 0.001); and from 9% to 51% for AC patients (p < 0.001). Surgical procedure varied significantly by demographic, socioeconomic status, facility, and tumor-related factors. 65% of cT1N0 cancers had concordant clinical and pathological staging after esophagectomy, with 11% having positive nodal disease; 44% were concordant after local excision. While no significant difference was seen in unadjusted survival, adjusted Cox Regression analysis indicated worse survival after esophagectomy compared to local excision for all cases (HR 1.67; 95% CI, 1.40-2.00) and for ACs with concordant staging (HR 1.54; 95% CI, 1.11-2.14). Conclusions: Local excision for cT1N0 esophageal cancer has increased over time. Staging concordance for esophagectomy is seen in two-thirds of cases. Contrary to our hypothesis, patients undergoing local excision for T1N0 cancers have better overall survival than those undergoing esophagectomy, which may reflect early differences in mortality and/or selection bias. As this study was unable to distinguish T1a from T1b, further analysis is warranted.
A prospective, multicenter, randomized clinical trial of UGT1A1 promoter polymorphism for irinotecan dose escalation in metastatic colorectal cancer patients treated with bevacizumab plus FOLFIRI as the first-line setting (PURE FIST).
Authors: Wang Jaw-Yuan
Published: March 2017

625

Background: The polymorphisms of genes encoding drug-metabolizing enzymes can play a crucial role in the increased susceptibility of patients to chemotherapy. This study is a prospective, multicenter, randomized clinical trial to compare the clinical outcomes and adverse events in metastatic colorectal cancer (mCRC) patients treated with bevacizumab plus 5-fluorouracil/leucovorin/irinotecan (FOLFIRI) with or without UGT1A1genotyping and irinotecan dose escalation as the first-line setting (NCT02256800). Methods: The enrolled patients were randomly assigned to one of two groups on the basis of receiving UGT1A1 genotyping or not. The study group receives a biweekly FOLFIRI plus bevacizumab, with irinotecan dose escalation based on UGT1A1 genotyping; whereas the control group receives the conventional biweekly FOLFIRI plus bevacizumab without UGT1A1 genotyping. The primary endpoint was the overall response rate (ORR), and the second endpoint was progression-free survival (PFS), overall survival (OS) and toxicities between the two groups. Results: BetweenAugust 2013 and May 2016, eighty-eight mCRC patients were enrolled, including 45 patients in the study group and 43 patients in the control group. With a median follow up time of 12.5 months (range, 5-30), the ORR was 71.4% vs. 44.2% ( P = 0.011). The PFS was 14.04 ± 1.44 vs. 9.08 ± 0.58 months in study group and control group ( P = 0.195), OS was 19.12 ± 1.24 vs. 14.16 ± 1.04 months ( P = 0.098), and ≥ Gr. III anemia was 2.2% vs. 23.3% ( P= 0.002) in study group and control group, respectively. However, there were no significant differences in ≥ Gr. III leukopenia or diarrhea even though the 1.36 fold relative dose intensity (RDI) in the study group. Conclusions: In the current study, patients with mCRC undergoing UGT1A1 genotyping may receive escalated doses of irinotecan for a potentially more favorable clinical response and outcome, in addition to acceptable toxicities. Such a personalized medicine based on genotyping may be feasible for potentially clinical practice. Clinical trial information: NCT02256800.
Tumor mutational burden of microsatellite stable metastatic colorectal tumors by patient factors and KRAS, BRAF, and PIK3CA mutation status.
Authors: Gong Jun , Sy Marvin , Fakih Marwan
Published: March 2017

627

Background: Microsatellite instability (MSI) can predict response to pembrolizumab in metastatic colorectal cancer (mCRC). Benefit to PD-1 blockade is limited in microsatellite stable (MSS) mCRC yet MSS tumors represent >95% of all mCRC cases. Evidence suggests that high mutational load remains an important predictor of benefit to checkpoint inhibition across several tumors. We aimed to explore associations between tumor mutational burden (TMB) and various patient factors and mutations of interest to identify features of patients (pts) with MSS mCRC of potential significance to prognosis and PD-1 targeting. Methods: We conducted a retrospective study based on next-generation sequencing (NGS) of MSS metastatic colorectal tumors at our single institution using FoundationOne. TMB was categorized as high ≥ 20 mutations/megabase (Mb), intermediate 6-19 mutations/Mb, and low ≤ 5 mutations/Mb. TMB frequencies were analyzed according to age (≤50, >50, >65, and >70 years old), gender, race, location of primary tumor, and KRAS, BRAF, and PIK3CA mutations. Results: Among 194 MSS tumors with available TMB data, frequencies of tumors with intermediate TMB were significantly different in pts ≤50 years (24.6%, n=69) than in pts >65 (48.9%, n=47, Fisher’s exact (two-tailed) p=0.0096) and >70 (53.1%, n=32, p=0.0067). Frequencies of tumors with low TMB were significantly different in pts ≤50 years (75.4%, n=69) than in pts >65 (46.8%, n=47, p=0.0029) and >70 (43.8%, n=32, p=0.0032). Frequencies of high TMB tumors by age were 0% (≤50 years), 1.6% (>50), 4.3% (>65), and 3.1% (>70) and did not reach significance. Frequencies of high, intermediate, or low TMB tumors were not significantly different based on gender, race, and location of primary tumor. Frequencies of KRAS, BRAF, and PIK3CA mutations were similar across all TMB categories (p=0.7995, 0.8774, and 0.053, respectively). Conclusions: Older individuals with mCRC are less likely to have a low mutation burden by FoundationOne assay. Further validation of these findings in larger cohorts may indicate a potential higher likelihood for elderly colorectal cancer patients to benefit from immune-based strategies.
The relationship between the non-canonical NF-κB pathway, tumour microenvironment, systemic inflammation and survival in patients undergoing surgery for colorectal caner.
Authors: Patel Meera , Bennett Lindsay , Quinn Jean A , van Wyk Hester Catharina , Horgan Paul G. , McMillan Donald C , Roseweir Antonia K , Park James Hugh , Edwards Joanne
Published: March 2017

631

Background: In patients with colorectal cancer (CRC) the local and systemic inflammatory responses (LIR and SIR) are important determinants of disease progression. The present study examines the association of transcription factor RelB, a key member of the non-canonical NF-κB pathway and its association with LIR and SIR in patients undergoing resection of CRC. Methods: Patients with stage I-III CRC who underwent curative resection in a single institution and were in a previously constructed tissue microarray were studied. IHC was utilised to examine cytoplasmic and nuclear RelB expression. The relationship between RelB, clinicopathological characteristics, LIR (Klintrup-Mäkinen (KM) grade, CD3+ and CD8+T-cell density), SIR and cancer-specific survival (CSS) was examined. Results: 208 patients were included in the analysis. Cytoplasmic RelB (cyto-RelB) was associated with nuclear RelB ( p=0.006). High expression of cyto-RelB was associated with MMR competence ( p=0.010) but not with TNM stage ( p=0.468), venous invasion ( p=0.973), tumour budding ( p=0.068), tumour necrosis ( p=0.786), tumour cell proliferation ( p=0.907), BRAF V600E mutation ( p=0.585) or administration of adjuvant chemotherapy ( p=0.853). High cyto-RelB was inversely associated with mGPS (mGPS >1: low cyto-RelB – 19% vs. high cyto-RelB – 8%, p=0.017). Also, cyto-RelB was inversely associated with tumour inflammatory cell infiltrate at the margin, Klintrup-Mäkinen grade ( p=0.059), (CD3+ p=0.010, CD8+ p=0.007) and in the tumour (CD3+ p=0.002) and a trend with tumour stroma percentage ( p=0.079). High expression of cyto-RelB was not significantly associated with CSS ( p=0.052). Conclusions: In patients undergoing CRC resection, high expression of cyto-RelB was associated with an adverse host local inflammatory response. Up-regulation of the non-canonical NF- κB pathway may be an important mechanism whereby the tumour deregulates local inflammatory responses and evades host immunosurveillance. Further investigation of inflammation based signal transduction pathways in patients with colorectal cancer is warranted.
Anti-KRAS siRNA nanoparticles for targeted colorectal cancer therapy.
Authors: Krasnick Bradley , Strand Matthew S. , Bi Ye , Goedegebuure Peter S. , Fleming Timothy , Wickline Samuel A. , Pan Hua , Fields Ryan C
Published: March 2017

636

Background: Standard treatment for metastatic colorectal cancer (mCRC) is systemic chemotherapy with anti-EGFR treatment, depending on KRAS mutational status. However, tumors harboring a KRAS mutation do not respond to existing targeted therapy. Moreover, targeting mutant KRAS has, to date, not been possible. Herein, we explore using a KRAS inhibitory nanoparticle (NP), to directly knock down mutant KRAS. Methods: Utilizing fluorescent-labeled small interfering RNA (siRNA) NPs, uptake was assessed via fluorescent microscopy. KRAS mutant CT26 and wild-type MC38 CRC cell lines were incubated with either scramble (Sc) sequence siRNA NP, KRAS siRNA NP, or FOLFOX (fluorouracil + oxaliplatin) chemotherapy ± KRAS siRNA NP. Cell viability was assessed via a luminescent viability assay. KRAS and cleaved caspase 3 protein expression were assessed using western blotting. Results: Fluorescent NP uptake was demonstrated in CT26 cells as early as 260 minutes post treatment, with increased uptake through 780 minutes. Decreased cellular viability was seen with KRAS siRNA NP treated CT26 cells, as compared to both Sc siRNA NP and non-treated CT26 cells (both p < 0.0001). Cell viability was significantly diminished with FOLFOX combined with KRAS siRNA NP as compared to FOLFOX alone for CT26 cells ( p = 0.0003), but not MC38 cells (p = 0.2259). Western blot demonstrated decreased KRAS and increased cleaved caspase 3 expression in CT26 cells treated with KRAS siRNA NP. Conclusions: A KRAS siRNA tagged NP was internalized by the CRC cells in vitro, and induced cellular death via apoptosis in mutant type KRAS CRC. In addition, KRAS siRNA NP acted synergistically with FOLFOX chemotherapy to enhance cell death. We believe KRAS inhibition based NP treatment is a promising target for mutant type KRAS CRC. [Table: see text]
Patterns of KRAS, NRAS and BRAF testing among patients with metastatic colorectal cancer in the EU5.
Authors: Lowe Kimberly , Niu Xin , Kafatos George , Kelsh Michael Anthony
Published: March 2017

637

Background: In mid-2013, the requirement for mCRC patients treated with panitumumab changed from wild-type KRAS to wild-type RAS ( KRAS and NRAS). The objective of this study was to describe patterns of KRAS, NRAS, and BRAF testing among mCRC patients in the EU5 (United Kingdom (U.K.), Germany, France, Italy and Spain). Methods: This descriptive study utilized Oncology Analyzer data (IMS Health Oncology, Fairfield, CT, USA), which provides real-world quarterly cross-sectional, epidemiologic and clinical data for mCRC patients. Trends in biomarker testing during the study period were evaluated by calculating the frequencies and proportions of patients who were /were not tested for combinations of KRAS, NRAS, and BRAF within each of the five countries. KRAS and BRAF data were available in all countries from quarter three (Q3) 2012 to quarter two (Q2) 2015. NRAS was available in all countries from quarter 1 (Q1) 2014 to Q2 2015. Frequencies and proportions were also calculated to summarize patient demographic and clinical characteristics between groups with different testing choices. Results: A total of 32,961 patients with mCRC were included in the analyses. Prior to 2013, the majority of patients in each country were tested for only KRAS. The proportion of patients tested for both KRAS and NRAS increased substantially during the period of data availability for NRAS (Q1 2014 to Q2 2015), including <1% to 19.1% in the U.K., 8.8% to 31.9% in France, 13.1% to 43.2% in Germany, 5.9% to 24.6% in Italy, and 8.7% to 31.4% in Spain. The proportion of patients who were tested for all three biomarkers also increased during the period of data availability for all three biomarkers (Q1 2014 to Q2 2015), including 2.3% to 16.4% in the U.K., 4.8% to 26.6% in France, <1% to 2.4% in Germany, 4.2% to 22.3% in Italy, and 1.5% to 9.3% in Spain. There were few differences in patient demographic or clinical characteristics between those with different testing groups. Conclusions: This analysis provides a description of biomarker testing using read-world data and suggests an overall increase in testing among mCRC patients in the EU5. Although there was a trend towards increased testing, many mCRC patients are still not tested for these important biomarkers.
Parallel sequencing of a 50 genes panel in metastatic colorectal cancer (MCRC) patients (pts) treated with intensive first-line FIr-B/FOx triplet chemotherapy plus bevacizumab (BEV): Preliminary data and clinical outcome.
Authors: Bruera Gemma , Malapelle Umberto , Pepe Francesco , Pisapia Pasquale , Dal Mas Antonella , Calvisi Giuseppe , Troncone Giancarlo , Ricevuto Enrico
Published: March 2017

644

Background: RAS/BRAF genotype guide MCRC treatment. First line triplet chemotherapy/BEV significantly improved PFS and OS. OS was significantly worse in KRAS c.35G > A and BRAF mutant (mt). Most CRC (86%) harbored mt genes, prevalently TP53, RAS, BRAF, PIK3CA. Methods: MCRC samples of 67 pts treated with FIr-B/FOx (77% overall) were analyzed through a 50 genes panel (PGM/Colon Lung Cancer) by ION Torrent: 57 (85%) primary, 10 (15%) metastatic samples; 59 (88%) pre-, 8 (12%) post-treatment. Molecular diagnostic criteria: > 50% coverage; > 1% mutant allelic fraction. Clinical outcomes were evaluated and compared by log-rank. Results: All wild-type (wt) and mt MCRC were 6 (8.9%) and 61 (91.1%), respectively; median mt genes 3 (1-12). Mt genes, 35 (%): KRAS 44 (65.6%), TP53 38 (56.7%), APC 26 (38.8%), KIT 23 (34.3%), PDGFRA 19 (28.3%), PIK3CA 18 (26.8%), EGFR and NRAS 15 (22.3%), SMAD4 10 (14.9%), FBXW7 and MET 7 (10.4%), GNAS and PTEN 6 (8.9%), BRAF and NOTCH1 5 (7.4%), ATM, PTPN11 and SMARCB1 4 (5.9%), HRAS, KDR, JAK3 and VHL 3 (4.4%), ERBB2, FGFR2, FGFR3, IDH1 and STK11 2 (2.9%), ABL1, AKT1, CDKN2A, FGFR1, FLT3, HNF1A, RB1, RET 1 (1.4%). BRAF mutations: c.1756G > A, 1796C > T, 1405G > A, 1406G > C. Median follow-up 21 months (m), overall PFS 13, OS 27m: KRAS exon 2 ( KRAS2) wt/mt, PFS 14/12m, OS 28/21m, respectively, not significantly different; c.35G > A KRAS mt showed trendly worse OS 14m; BRAF mt significantly worse PFS (8 vs 14m, p .026) and OS (11 vs 28m, p .002); RAS2-4/ BRAF15 wt/mt, PFS 18/13m (p .954), OS 28/22m (p .956). TP53 wt/mt, PFS 14/12m, OS 28/23m. All wt, PFS 24, OS 44m, not significantly different vs ≥ 1 mt gene. Conclusions: Multigenic analysis of MCRC patients treated with FIr-B/FOx shows worse clinical outcome conferred by uncommon BRAF mutations.
Prognostic impact of preoperative albumin to globulin ratio in curative colon cancer patients.
Authors: Toiyama Yuji , Fujikawa Hiroyuki , Inoue Yasuhiro , Imaoka Hiroki , Okigami Masato , Yasuda Hiromi , Hiro Junichiro , Yoshiyama Shigeyuki , Saigusa Susumu , Kobayashi Minako , Ohi Masaki , Araki Toshimitsu , Mohri Yasuhiko , Kusunoki Masato
Published: March 2017

647

Background: Albumin to globulin ratio (AGR) has been reported to predict long term mortality in patients with several cancers. However, prognostic impact of preoperative AGR in colon cancer patients with curative intent has not yet been fully addressed. Therefore, we, for the first time, investigated the association between AGR and clinico-pathological findings including overall survival (OS) and disease free survival (DFS) in stage I-III colon cancer patients. Methods: Clinicopathological findings including preoperative laboratory data (carcinoembryonic antigen [CEA] and AGR) from 251 curative colon cancer patients were assessed as indicators of early recurrence and poor prognosis in this retrospective study. AGR was calculated as [AGR = albumin/ (total protein - albumin)]. The cut-off value of AGR was 1.32 in current study. Results: Several clinicopathological categories related with tumor progression such as lymph node metastasis, T4 tumor, large tumor size, undifferentiated tumor, venous and lymphatic invasion, and high CEA were significantly associated with low AGR level. The patients with low AGR were significantly poorer OS (P = 0.001) and DFS (P = 0.003) than those with high AGR, respectively. In addition, multivariate analyses demonstrated that low AGR was independently associated with early recurrence (HR = 2.87, P = 0.007) and poor prognosis (HR = 2.56, P = 0.008), respectively. On the other hand, sub analysis of survival curves revealed that stage III colon cancer patients with low AGR were significantly poorer OS (P = 0.007) and DFS (P = 0.02) than those with high AGR, respectively. Furthermore, significantly poorer OS and DFS were also shown in stage I-II colon cancer patients with low AGR, respectively (OS: P = 0.02, DFS: P = 0.01). Conclusions: Preoperative AGR was an independent predictor of early recurrence and poor prognosis in curative colon cancer patients. AGR may represent a simple, potentially useful predictive biomarker for selecting stage I-II colon cancer patients who might need adjuvant chemotherapy. Furthermore, AGR may select candidates who are better to introduce more intensive adjuvant chemotherapy after curative operation in stage III colon cancer patients.