bluuebunny/crossref_metadata_2025 huggingface dataset:
Single nucleotide polymorphisms in miRNA binding sites of nucleotide excision repair-related genes to predict clinical benefit of oxaliplatin in FOLFOXIRI plus bevacizumab in TRIBE trial.
Authors: Suenaga Mitsukuni , Schirripa Marta , Cao Shu , Zhang Wu , Yang Dongyun , Cremolini Chiara , Murgioni Sabina , Lonardi Sara , Ning Yan , Okazaki Satoshi , Berger Martin D , Miyamoto Yuji , Gopez Roel , Barzi Afsaneh , Loupakis Fotios , Falcone Alfredo , Lenz Heinz-Josef
Published: January 2019

663

Background: Nucleotide excision repair (NER) is known as part of the DNA damage response (DDR) against platinum-induced DNA damage. Polymorphisms in miRNA binding sites of replication protein A ( RPA2) and General Transcription Factor IIH Subunit 1 ( GTF2H1) genes in NER pathway have shown to correlate with colorectal cancer risk. We tested whether the NER-related single nucleotide polymorphisms (SNPs) predict oxaliplatin efficacy in FOLFOXIRI regimen in patients (pts) with metastatic colorectal cancer (mCRC). Methods: Genomic DNA was extracted from 457 blood samples of mCRC pts enrolled in the TRIBE trial: 230 pts received first-line (1L) FOLFOXIRI plus bevacizumab (discovery cohort: median age, 60 yrs; median follow-up, 46.6 mos); and 227 pts received 1L FOLFIRI plus bevacizumab (control cohort: median age, 60 yrs; median follow-up, 49.0 mos). SNPs were analyzed by PCR-based direct sequencing. Progression-free survival (PFS) and overall survival (OS) were analyzed using Kaplan-Meier curves, log-rank test, and Cox proportional hazards regression. Results: In KRAS wild-type population among the discovery cohort, the C/C variant in RPA2 rs7356 was associated with shorter PFS and OS than any T allele (9.1 vs 13.3 mos, HR 2.32, 95%CI: 1.07-5.03, P= 0.020; 20.8 vs 37.1 mos, HR 1.94, 95%CI: 0.94-3.99, P= 0.059). Any C allele in GTF2H1 rs4596 was associated with shorter PFS than the G/G variant (10.8 vs 13.3 mos, HR1.64, 95%CI: 0.95-2.81, P= 0.045). In multivariable analysis, RPA2 rs7356 remained significant for PFS and OS (HR 2.97, 95%CI: 1.27-6.94, P= 0.012; HR 2.58, 95%CI: 1.19-5.58, P= 0.016). In all population of the discovery cohort, pts with any GTF2H1 rs4596 C allele had remarkably shorter PFS and OS than those with the G/G variant (11.3 vs 13.2 mos, HR 1.32, P= 0.087; 28.6 vs 34.3 mos, HR 1.36, P= 0.076). The findings remained in multivariable analysis. These findings were not confirmed in the control cohort. Conclusions: We suggest that miRNA binding sites of NER-related genes regulate DDR for oxaliplatin-related DNA damage. Germline RPA2 and GTF2H1 polymorphisms may predict oxaliplatin efficacy in mCRC pts.
Comparison of the real-world adherence and compliance patterns with trifluridine/tipiracil (FTD/TPI) and regorafenib (REG) for the treatment of metastatic colorectal cancer (mCRC).
Authors: Patel Anuj K. , Duh Mei Sheng , Barghout Victoria , Yenikomshian Mihran Ara , Xiao Yongling , Wynant Willy , Tabesh Majid , Fuchs Charles S.
Published: January 2019

666

Background: FTD/TPI and REG both prolong survival in refractory mCRC and have similar indications with different side effect profiles. This study compares real-world treatment patterns with FTD/TPI and REG for mCRC in a large, representative US claims database. Methods: Retrospective data from 10/2014 to 7/2016 from the US Symphony Health Solutions’ Integrated Dataverse (IDV®) database were analyzed for patients receiving FTD/TPI or REG. The index date was the date of first FTD/TPI or REG prescription. Patients were included if: 1) age ≥18 years old, 2) ≥1 CRC diagnosis, 3) no diagnosis of gastric cancer or gastrointestinal stromal tumor, and 4) continuous clinical activity for ≥3 months before and after index date. The observation period spanned from index date to end of data, end of continuous clinical activity, or switch to another mCRC treatment. Adherence was assessed using medication possession ratio (MPR) ≥0.80 and proportion of days covered (PDC) ≥0.80 at 3 months. Compliance was assessed using time to discontinuation over the observation period using allowable gaps of 45, 60, or 90 days. Patients who never discontinued therapy were censored at the end of the observation period. Outcomes were compared between FTD/TPI and REG using multivariate logistic regression and Cox proportional hazards models, adjusting for demographic and clinical baseline characteristics. Results: A total of 1,630 FTD/TPI patients and 1,425 REG patients were identified. Mean ± standard deviation (SD) age of FTD/TPI patients was 61.0 ± 11.0 compared to 62.8 ± 10.9 for REG patients (p < 0.001). FTD/TPI patients were 80% more likely to have a MPR ≥0.80 compared to those on REG (Odds Ratio [OR] = 1.80, p < 0.001) and more than twice as likely to have a PDC ≥0.80 (OR = 2.66, p < 0.001) at 3 months. FTD/TPI patients were 37% less likely to discontinue their treatment compared to those on REG when using gaps of 60 days (Hazard Ratio = 0.63, p < 0.001). Similar results were found with 45 and 90 days. Conclusions: In this retrospective study of mCRC patients, patients on FTD/TPI were significantly more likely to adhere and comply with therapy compared to those on REG.
Lyc-55716: A novel small-molecule RORγ agonist immuno-oncology agent: Rationale for tumor selection and clinical evaluation of gastric and esophageal carcinoma in phase 2a expansion.
Authors: Weems Garry Alan , Hu Xiao , Liu Xikui , Li Hongxiu , Bogdan Madhumita , Gao Yilin , Fox Brian , Wilkins H. Jeffrey , Carter Laura
Published: January 2019

67

Background: Type 17 effector T cell differentiation and function are regulated by the master transcription factor retinoic acid receptor–related orphan receptor γt (RORγ). Synthetic RORγ agonists, which modulate immune cell gene expression to enhance effector functions and decrease Treg and checkpoint pathways, have shown promise as mono- and combination therapy in syngeneic tumor models. Translational and bioinformatics assessments were performed during Phase 1 testing of the investigational RORγ agonist LYC-55716 to support inclusion of patients with gastric carcinoma (GC) and esophageal carcinoma (EC) in a Phase 2a expansion of clinical trial LYC-55716-1001 (NCT02929862). Methods: Transcriptional profiling of murine and human T cells treated ± RORγ agonists identified a gene signature. Using The Cancer Genome Atlas (TCGA), and other public datasets, data on GC and EC patients were evaluated to determine: (a) RORγ and RORγ-inducing cytokine expression; (b) signature genes associated with RORγ biology, surrogate biomarkers for endogenous RORγ ligands, and correlations with prognosis; (c) tumor microenvironment (TME) immune profiles. Results: Expression: In TCGA RNA sequencing data analysis, 50%-70% of GC and EC samples expressed moderate to high levels of RORγ, indicating infiltration of Type 17 T cells into the tumors. Furthermore, genes that support expression of RORγ were highly expressed in ~75% of GC and EC tumors. Biology: TCGA data showed that GC and EC tumors express low levels of sterol efflux genes, suggesting low levels of endogenous agonists in the TME. In analysis of public datasets, expression of the RORγ target gene IL17A correlated with a favorable prognosis for EC. In addition, some of the other RORγ signature genes also correlated with good prognosis in GC and EC. Immune profile: TCGA showed infiltration of T cells and a high mutational burden in GC and EC, which are positively associated with efficacy of immunotherapy. Conclusions: Bioinformatics assessments of RORγ expression, biology, and tumor immune profiles support the inclusion of GC and EC patients in an ongoing Phase 2 clinical trial of LYC-55716.
Circulating angiogenesis-related markers as predictors of benefit from regorafenib in metastatic colorectal cancer (mCRC) patients (pts).
Authors: Antoniotti Carlotta , Marmorino Federica , Pennati Marzia , Zaffaroni Nadia , Rossini Daniele , Borelli Beatrice , Zucchelli Gemma , Moretto Roberto , Pietrantonio Filippo , Masi Gianluca , Vannini Francesca , Colombo Camilla , Gini Serenella , Saettini Alessandra , Delliponti Laura , Brunetti Isa Maura , Pfanner Elisabetta , Falcone Alfredo , Cremolini Chiara
Published: January 2019

675

Background: Regorafenib is a treatment option for refractory mCRC pts. Considering its limited clinical benefit and the palliative setting, patients’ selection is essential to optimize the cost-effectiveness ratio. We aimed to describe the modulation of selected circulating angiogenic factors by regorafenib and to investigate their correlation with clinical outcome. Methods: IL-8, Ang-2, PDGF, bFGF, Tie-2, sVEGFR2, sVEGFR3, PlGF, VEGF-A, VEGF-B were assessed by ELISA on plasma samples collected at baseline (d1), after 15 days of treatment (d15), at the best RECIST response, at disease progression (PD) in mCRC pts treated with regorafenib, as per indication. Comparisons among concentrations of each marker at different timepoints were performed by Wilcoxon test. Markers showing significant changes were analyzed for correlation with outcome. Results: 105 pts were included. Median PFS and OS were 2.1 and 7.0 months (mos), respectively. As compared to d1, IL-8 and Ang-2 levels increased at PD. An early decrease at d15 was observed for PDGF, Tie-2, sVEGFR2 and sVEGFR3 levels, followed by an increase at PD. Conversely, PlGF levels increased at d15 and then decreased at PD. Baseline levels of Ang-2 and Tie-2 below the median value were associated with longer PFS (HR:0.61 [0.37-0.83] p = 0.006; HR:0.69 [0.43-0.95] p = 0.04, respectively) and OS (HR:0.45 [0.26-0.60] p < 0.0001; HR:0.68 [0.44-0.98] p = 0.04, respectively). With regard to Ang-2, 40 (45%) out of 89 pts with available plasma samples at d15 showed increased levels at d15 as compared to d1. Among them, 21 (53%) achieved disease control, as compared to 14 out of 49 (29%) pts with Ang-2 decreased levels (p = 0.03). Median PFS of pts with increased and decreased Ang-2 levels were 3.1 and 1.8 mos, respectively (HR:0.57 [0.33-0.78] p = 0.004). Conclusions: A dynamic modulation of plasma angiogenic factors occurs during the treatment with regorafenib. Low baseline Ang-2 and Tie-2 levels seem to be associated with good prognosis. The early modulation of Ang-2 levels may predict benefit from regorafenib. These results need validation in independent series.
ALK alteration association with prognosis in metastatic colorectal cancer: A single institution retrospective cohort study.
Authors: Kasi Anup , Chidharla Anusha , Schmitt Sarah , Pessetto Ziyan Y. , Gupta Nikhil , Paluri Ravi Kumar , Saeed Anwaar Mohammed , Godwin Andrew K.
Published: January 2019

676

Background: The Anaplastic Lymphoma Kinas (ALK) gene is altered in various malignancies which can be targeted with several inhibitors. However, the incidence of mutations in ALK are not well defined in colorectal cancers (CRC). We aim to study the prevalence of ALK mutations in metastatic CRC (mCRC) patients and correlate with clinical outcomes. Methods: We retrospectively evaluated 54 mCRC patients treated with chemotherapy at Kansas University Cancer Center between June 2010 and September 2017 who underwent our inhouse targeted 17 gene next-generation sequencing test using an illumina MiSeq technology. The panel included AKT1, ALK, BRAF, EGFR, FLT3, HRAS, IDH1, IDH2, JAK2, KIT, KRAS, NPM1, NRAS, PDGFRA, PIK3CA, PTEN, and TP53 genes. We compared baseline characteristics, tumor location (left vs right) and clinical outcomes between patients with ALK mutations versus ALK wild type. Results: We retrospectively evaluated 54 mCRC patients treated with chemotherapy at Kansas University Cancer Center between June 2010 and September 2017 who underwent our inhouse targeted 17 gene next-generation sequencing test using an illumina MiSeq technology. We compared baseline characteristics, tumor location (left vs right) and clinical outcomes between patients with ALK mutations versus ALK wild type. Results: We found 10 pts (18.5%) with ALK mutations and compared with 44 patients that were ALK wild type. Demographic and baseline characteristics (ALK mutation/wild type) were as follows: Median age 61 vs 61 years, Gender (male): 80% vs 50%, Left sided tumor 70% vs 60%. Of 41 evaluable patients (ALK mutation vs wild type), Objective response rate (stable + partial response) was 50% vs 91% [p = 0.03], 6-month PFS 33% vs 77%, Median PFS 12 months vs 17.7 months [p = 0.2], Median OS was 14.5 months vs not reached [HR 28.98, p = 0.025]. Conclusions: Patients harboring ALK mutations had a statistically significant poorer objective response rate and OS compared to ALK wild type patients. Findings of our study highlight the importance of assessing ALK gene alteration as a prognostic biomarker for mCRC. Therefore ALK mutation appears to be poor prognostic factor and needs to be validated in larger cohorts.
Therapeutic relevance of homologous recombination repair (HRR) pathway variants in metastatic colorectal cancer (mCRC).
Authors: Narra Ravi Kishore , Singavi Arun K. , Ritch Paul S. , Thomas James P. , Alqwasmi Abdel , Szabo Aniko , George Ben
Published: January 2019

677

Background: Intact HRR pathway genes play a critical role in repairing double-stranded DNA breaks. Genomic alterations (GA) in several HRR pathway genes have been well characterized, demonstrating prognostic and predictive significance. However, there are many variants of unknown significance (VUS) in the HRR pathway genes that need better characterization. Methods: Patients (pts) with mCRC that harbored VUSs in HRR pathway genes (HRR- VUS; BRCA1/2, ATM, ATR, RAD50, RAD51, PALB2, CHEK1, CHEK2) and treated with front line platinum therapy were identified by review of our institutional molecular data base (MDB) and EMR. Time to 2nd line therapy (TT2L) and Overall Survival (OS) were calculated. DNA was extracted from formalin fixed paraffin embedded clinical specimens and Next Generation Sequencing (NGS) was performed on hybrid-capture, adaptor ligation based libraries to a mean coverage depth of > 600 unique reads utilizing the Foundation Medicine NGS platform. Results: Among the 873 pts in our institutional MDB, 96 (11%) had mCRC, 20 (21%) harbored HRR pathway GAs, 15 (16%) were categorized as VUSs. HRR-VUS pts had a median age of 57 at diagnosis, 5 (33%) were male, 3 (20%) were right-sided. Distribution of HRR-VUSs in this cohort is summarized in table 1. All pts with HRR-VUSs were micro-satellite stable (MSS), median TMB was 4.5 (range 0 – 8.1). Median TT2L and OS in mCRC pts with HRR-VUSs were 8.0 and 43 months (m) respectively. Two pts with HRR-VUSs had TT2L > 24 m and OS > 40 m; one of them had a BRCA1P1464A mutation, while the other had a BRCA2 amplification. Conclusions: BRCA2, ATM and BRCA1 were the HRR genes that harbored VUSs most frequently in pts with mCRC. Majority with HRR-VUSs had left sided primaries, were MSS and had low TMB. BRCA1P1464A mutation in the ATM binding domain needs further characterization on account of prolonged TT2L with platinum based chemotherapy. Patients with HRR-VUSs that are predictive of benefit with platinum based chemotherapies could be considered for maintenance therapy with PARP inhibitors [Table: see text]
Molecular rationale for ERK and EGFR inhibition in colorectal cancer.
Authors: Yeatman Timothy Joseph , Davis Thomas , Yang Mingli , Schell Michael J. , Wang Heiman , Rocha Lima Caio Max S. , Pledger Jack
Published: January 2019

679

Background: Mutational activation of the RAS pathway occurs in >50% of human CRC tumors, and to date has been difficult to overcome. We recently identified receptor tyrosine phosphatase-S (PTPRS) as an important regulator of the RAS pathway, which is mutated in ~11% of CRC. Methods: Global gene expression and DNA sequencing data (1321 cancer genes) were integrated across 468 CRC tumors to identify PTPRS as one of the most commonly mutated genes with a high correlation to a RAS activation score. Apoptotic responses to inhibitors of ERK, EGFR and their combination were tested in isogenic wild-type and mutant RAS cell lines where PTPRS had been knocked out by CRISPR. Results: PTPRS was in the top 15 genes correlating with RAS pathway activation scores; as expected, KRAS, BRAF were the most correlated genes. In order to determine the effect of the loss of PTPRS on RAS/ERK signaling, we used CRISPR to knockout PTPRS in an isogenic pair of CRC cell lines (HCT116) with mutant or wild type KRAS. Cell lines lacking PTPRS expression had elevated phospho-ERK activity. Interestingly, cell lines with PTPRS knockout and elevated ERK phosphorylation had an increased sensitivity to ERK inhibitors. Moreover, the loss of PTPRS allowed ERK to localize to the nucleus. Apoptosis increased dramatically from 13.1% in control cells to 53.8% in PTPRS KO cells treated with both ERK and EGFR inhibitors (Table), a synergistic effect not seen in wild type KRAS cells, where only ERK inhibitors were active. Conclusions: The loss of PTPRS allows increased activation of ERK (and the RAS pathway) which is associated with ERK addiction and increased sensitivity to ERK inhibitors. Surprisingly, ERK inhibitors synergize with EGFRi in wild type KRAS cell lines absent PTPRS. These data suggest the potential for mutant PTPRS to serve as a biomarker for ERK/EGFR inhibition. [Table: see text]
Effect of ASXL1 on the stemness of colorectal cancer initiating cells.
Authors: Isobe Taichi , Zarnegar Mark A. , Abdel-Wahab Omar Ibrahim , Clarke Michael F.
Published: January 2019

683

Background: Impaired cellular differentiation causes uncontrolled proliferation of stem cells and can be a cause of oncogenesis. The epigenome regulates the expression of genes that are essential for cellular differentiation. On the other hand, chromosomal abnormalities are common in colorectal cancer cells. We hypothesize that epigenetic disorders caused by chromosomal abnormalities may result in oncogenesis. Methods: The Cancer Genome Atlas (TCGA) database was investigated to identify epigenetic regulating genes affected by chromosomal abnormalities. Patient-derived colorectal cancer cells were obtained from consented patients to validate these findings. Results: The amplification of Chr20q11 is the most frequent chromosomal alteration in left-sided colonic and rectal cancers. ASXL1, a recruiter of PRC2, is coded in Chr20q11 and has upregulated expression in many cases. Excessive expression of ASXL1 may cause oncogenesis as PRC2 trimethylates histone H3 on lysine 27 (H3K27me3), an epigenetic mark of transcriptional silencing. Immunofluorescence revealed that the ASXL1 protein was located in the nuclei of colorectal cancer cells that did not express differentiation markers. This finding was consistent with the finding that ASXL1 highly expressed at the bottom of crypts where undifferentiated cells exist in normal colonic epithelium. Knockdown of ASXL1 in colorectal cancer cells reduced the levels of H3K27me3 and diminished organoid formation in vitro. Knockdown of ASXL1 in patient-derived cancer cells obtained from clinical specimens impaired the tumorigenicity of cancer initiating cells in vivo and increased the expression of differentiation markers. Murine colonic tumor cells from ApcMin mice highly expressed ASXL1, while homozygous Asxl 1 knockout reduced the occurrence of tumors in vivo. Many putative genes regulated by ASXL1 are identified by gene expression analysis and ChIP-sequencing. Conclusions: High expression of ASXL1 initiates oncogenesis and maintains the stemeness of colorectal cancer cells.
CEA as a pharmacodynamic biomarker for metastatic anal cancer.
Authors: Hester Robert Harrison , Taniguchi Cullen M. , Holliday Emma Brey , Koay Eugene Jon , Das Prajnan , Minsky Bruce D. , Rashid Asif , Mehdizadeh Amir , Ohinata Aki , Messick Craig , You Y. Nancy , Chang George J. , Bednarski Brian , Rodriguez-Bigas Miguel A. , Skibber John Michael , Wolff Robert A. , Eng Cathy , Morris Van Karlyle
Published: January 2019

684

Background: While carcinoembryonic antigen (CEA) as a tumor marker is frequently used in the management of colorectal adenocarcinoma, its clinical utility in squamous cell carcinoma of the anus (SCCA) has never been reported. Indeed, no tumor markers have been validated for monitoring tumor burden in the course of any HPV-associated malignancy like SCCA. We hypothesized that CEA levels might be preferentially elevated in patients with metastatic SCCA. Methods: Charts from 212 patients with SCCA were reviewed under an IRB-approved protocol for correlations between CEA levels and corresponding oncologic status. Clinical status was categorized as newly diagnosed non-metastatic (D), after chemoradiation with no evident remnant disease (N), recurrent/resectable SCCA (R), locally advanced/unresectable SCCA (U), metastatic SCCA to lymph nodes only (M-LN), or metastatic SCCA to visceral organs (M-V). Mean CEA levels were compared between subgroups via student t-tests, and frequencies of elevated CEA were compared via Chi-squared analyses. Results: 118 SCCA patients had metastatic disease (98 M-V, 19 M-LN). Mean CEA levels by clinical status were 5.4 (D), 2.0 (N), 2.3 (R), 4.6 (U), 6.0 (M-LN), and 22.2 (M-V), with a higher (statistically insignificant) mean CEA level in the M-V relative to other populations. However, patients with visceral metastases were more likely to have an elevated CEA at presentation (40.4%) relative to patients with newly diagnosed, non-metastatic SCCA (17.6%, p = .07) or recurrent SCCA (11.1%, p = .02). For patients with metastatic SCCA, a significant association existed between change in CEA and corresponding change in radiographic tumor dimensions (OR 24, p < 0.0001). Conclusions: Trends in CEA correlate with dynamic changes in tumor burden for patients with metastatic SCCA, and patients with metastatic SCCA were more likely to have an elevated CEA. Given that immune checkpoint blockade agents like nivolumab have proven benefit for metastatic SCCA, these data provide rationale for use of newer generation immunotherapeutic approaches like CEA-T cell bispecific antibodies, which target CEA-expressing tumors, in clinical trials for patients with metastatic SCCA.
Modulation of T cell and macrophage tumor infiltration by the TLR9 agonist lefitolimod in a murine model of colorectal cancer.
Authors: Schmidt Manuel , Oswald Detlef , Volz Barbara , Wittig Burghardt , Kapp Kerstin
Published: January 2019

687

Background: The use of TLR9 agonists for immunotherapeutic approaches is supported by preclinical and ongoing clinical studies. Lefitolimod, a covalently-closed dumbbell-shaped DNA construct, is an immune surveillance reactivator which broadly activates cells of the innate and adaptive immune system. The conversion of non-immunogenic (cold) tumors into immunogenic (hot) tumors, characterized by their T-cell infiltration, is a pre-requisite for response to immunotherapies. By its mode-of-action lefitolimod likely provides the necessary signals for activation of immune cells and their differentiation into anti-tumor effector cells as well as their recruitment to the tumor microenvironment (TME). Methods: To support the further clinical development of lefitolimod in mCRC, the syngeneic colon carcinoma model CT26 was used to evaluate anti-tumoral efficacy of lefitolimod and its modulation of the TME. The phenotype of T cells and macrophages present within tumor-infiltrating immune cells were analyzed by flow cytometry. Their localization within the tumor was investigated by immunohistochemistry. Results: Intratumoral injection of single-agent lefitolimod in the CT26 model resulted in reduced tumor growth. This was accompanied by an increased infiltration of T cells, in particular CD8+ T cells, into the tumor center. The CD8+ T cells were activated and showed an up-regulation of the cytolytic effector Granzyme B. Correspondingly, the ratio of activated CD8+ T cells with cytolytic function to regulatory T cells was increased by lefitolimod. Moreover, lefitolimod led to the recruitment of macrophages into the tumor and, importantly, an increase of M1 macrophages and a decrease of M2 macrophages inside the TME. Conclusions: Lefitolimod treatment resulted in modulation of the TME towards an increase of effector cells with anti-tumoral functions like cytotoxic CD8+ T cells and M1-type macrophages as well as a decrease of pro-tumoral M2-type macrophages. Therefore, lefitolimod provides the essential requirements for clinical response to immunotherapeutic drugs like checkpoint inhibitors. This predestines lefitolimod as combination partner in immuno-oncological trials.
Effect of tumor progression on colon cancer stem cell plasticity.
Authors: Chiu Vi Kien , Gallupalli Rama , Osman Diaa , Zeng Zhaoshi , Shia Jinru , Paty Philip , Le Rolle Anne-France
Published: January 2019

688

Background: Stage I colon cancer initiation is optimized through the activation of the embryonic stem cell (SC)-like program in colon adenoma cells (Le Rolle AF, et al., 2016). This malignant transformation requires cellular dedifferentiation since the embryonic SC-like program does not exist a priori in colon cells. High initial tumor plasticity provides a competitive advantage as the tumor has more intrinsic phenotypic flexibility to survive environmental challenges. Inhibition of the embryonic SC-like program represents a novel therapeutic strategy. Herein, we examine the evolution this high tumor plasticity in stage I-IV colon cancer. Methods: Human colon cancer tissues were collected prospectively under an MSKCC IRB protocol (Jan 1990-Dec 2000). RNA in situ hybridization of LGR5, a colon SC marker, was carried out on human colon tumors before and after therapeutic interventions. We performed Gene Set Enrichment Analysis (GSEA 2.0 Broad Institute software) using Affymetrix U133A expression profiles from normal colon mucosa (n = 33) and colon cancer epithelia from stage I (n = 17), II (n = 35), III (n= 39) and IV (n= 46) tumors. Statistical analyses were conducted with GraphPad Prism 5 and Microsoft Excel. Results: Putative colon SC markers and differentiation markers are increased and decreased, respectively, in stage I-IV colon cancer in comparison to normal colon. Although colon adenoma originates from LGR5+ colon SC, LGR5+ overexpression per se correlates with good prognosis stage IV colon cancer and is not a colon cancer stem cell biomarker. Maximal embryonic SC-like program enrichment occurs in stage I colon cancer. GSEA comparisons of moderately differentiated stages II-IV versus stage I colon cancer reveal progressive tumor differentiation from the stage I embryonic SC-like program toward the intestinal SC program at more advanced tumor stages. Notably, poorly differentiated stage IV colon cancer retains an embryonic SC-like program. Conclusions: We conclude that except for poorly differentiated tumors, colon cancer progression from stage I to IV involves a heterogeneous tumor differentiation process from an embryonic SC-like program towards the intestinal SC or more differentiated intestinal cell programs.
Clinical and molecular characterization of early-onset colorectal cancer patients with inflammatory bowel disease.
Authors: Willauer Alexandra Nicole , Loree Jonathan M. , Pereira Allan Andresson Lima , Lam Michael , Raghav Kanwal Pratap Singh , Morris Van Karlyle , Menter David , Shaw Kenna Rael , Broaddus Russell , Meric-Bernstam Funda , Wang Yinghong , Overman Michael J. , Kopetz Scott
Published: January 2019

689

Background: Recent reports suggest colorectal cancer incidence is steadily increasing in adults < 50 years old. This population has a number of unique pre-disposing conditions, such as inflammatory bowel disease (IBD), that may alter the disease biology within these patients. Methods: Using a cohort of 1876 patients with metastatic CRC (mCRC), we identified 622 patients aged < 50 years and reviewed their medical records for a diagnosis of IBD. Clinical, pathologic and overall survival (OS) were compared between patients with and without IBD in this age group. Results: Twenty of 622 (3.2%) early-onset mCRC patients had IBD. These patients were more likely to have mucinous or signet ring histologic features (50.0% vs 15.6%, OR 5.43, 95% CI 2.20-13.41, P < 0.0001) and less likely to have APC mutations (15.0% vs 42.7%, P = 0.019). A trend toward an increased prevalence of right-sided location (40.0% vs 24.1%, P = 0.053) and TP53 mutations (85.0% vs 65.1%, P = 0.091) and decreased prevalence of PIK3CA mutations (0.0% vs 15.5%, P = 0.057) and KRAS mutations (35.0% vs 50.0%, P = 0.11) were noted. There were no differences in prevalence of BRAF V600 (P = 0.99), CTNNB1 (P = 0.99), NRAS (P = 0.99), PTEN (P = 0.43), or SMAD4 (P = 0.75) mutations between patients with IBD and other early-onset mCRC. The presence of IBD in early-onset mCRC patients was associated with a worse OS compared to patients without IBD but who had mCRC (HR 1.82, 95% CI 0.90-3.69, P = 0.026). Conclusions: Early-onset mCRC patients with IBD are significantly more likely to have mucinous or signet ring histologic features and have fewer mutations in canonical CRC genes such as KRAS, PIK3CA, and APC. This suggests that this population likely develops through a unique molecular pathogenesis and they may respond differently to therapy. Further molecular characterization of this population is required.
Occurrence and characteristics of hospitalizations during first-line chemotherapy among individuals with metastatic colorectal cancer.
Authors: Fisch Michael Jordan , Grabner Michael , Mytelka Daniel S , Raval Amit D , Bowman Lee , Kern David M , Churchill Collin , Singer Joseph , Wetmore Stewart , Barron John , Eleff Michael
Published: January 2019

691

Background: Choosing chemotherapy for metastatic colorectal cancer (mCRC) requires balancing clinical effectiveness and risk of complications. This study characterized real-world inpatient/ER hospitalizations (HOSP) during first-line chemotherapy among individuals with mCRC. Methods: We conducted a retrospective cohort study of adults with mCRC identified using claims data from the HealthCore Integrated Research Environment as initiating first-line chemotherapy from 12/23/2013 to 06/30/2016 (no minimum follow-up). Cohorts were analyzed in aggregate and for the most frequently observed first-line agents (5 overlapping subcohorts). HOSPs were identified from initiation of first-line chemotherapy to 30 days after the end of first-line chemotherapy or last available data. Results: A total of 717 individuals (mean age 55y; 58% male; 44%/39%/6%/12% with ECOG = 0/1/2+/missing; median follow-up 116 days) met study criteria. Metastasis was most commonly to the liver (51%) and 53% of patients had cancer-attributable morbidities. Chemotherapies included 5-FU (79%), oxaliplatin (67%), bevacizumab (58%), irinotecan (21%), and capecitabine (19%). Overall, 40% of patients had ≥1 HOSP [n = 285; total 415 events], ranging from 38% to 49% across the 5 chemotherapy-based subcohorts; 12% (n = 85) had > 1 HOSP. The median time to first HOSP for patients with an event was 52 days. The median length of inpatient stays was 4 days; Infections/neutropenia (21%), bowel-related complications (17%), cardiac and circulatory disorders (9%), malnutrition (5%), pain (5%) and renal disease (2%) were the most common issues associated with inpatient HOSPs. An increase in HOSPs was observed with worsening ECOG status: 0 (34%), 1 (46%), and 2+ (65%). In regression analyses, ECOG≥1 was associated with a 64%-72% increase (p < 0.01) in the odds of HOSPs compared to patients with ECOG = 0. Conclusions: Approximately 40% of mCRC patients had hospitalizations during the study period. Hospital stays were typically short and associated with infections, neutropenia, or bowel-related complications. Further research is needed to determine how many of these hospitalizations may be avoidable.
Matrix metalloproteinase-related gene polymorphisms to predict efficacy of regorafenib in patients with metastatic colorectal cancer.
Authors: Suenaga Mitsukuni , Schirripa Marta , Cao Shu , Zhang Wu , Yang Dongyun , Ning Yan , Bergamo Francesca , Zagonel Vittorina , Rossini Daniele , Borelli Beatrice , Okazaki Satoshi , Berger Martin D , Miyamoto Yuji , Gopez Roel , Barzi Afsaneh , Yamaguchi Toshiharu , Loupakis Fotios , Lenz Heinz-Josef
Published: January 2019

692

Background: Matrix metalloproteinases (MMPs) are proteolytic enzyme for extracellular matrix and involved in regulation of tumor environment including angiogenesis. MMP-2 and MMP-9 were reported to be prognostic marker of anti-angiogenic therapy. We tested whether genetic polymorphisms in the MMP family predict clinical outcomes in patients with refractory mCRC receiving regorafenib. Methods: 228 patients with mCRC receiving regorafenib were included in this pooled analysis combining two different cohorts from Italy and Japan (median age, 62 years; male, 51.5%; median follow-up time, 26.2 months; Italian/Japanese, 150/79). Six single nucleotide polymorphisms (SNPs) of five genes in MMPs ( MMP2, MMP9, MMP14, TIMP2, CD44) were analyzed by PCR-based direct sequencing. Correlations between candidate SNPs and progression-free survival (PFS) and overall survival (OS) were analyzed using Kaplan-Meier curves and log-rank test. Disease control was compared by Fisher’s exact test. Multivariable analyses were conducted using the Cox proportional hazard regression model. Results: In univariate analysis, patients carrying the G/G variant in MMP2 rs1477017 had a significantly shorter OS compared to those with any A allele (4.4 vs. 7.8 months, HR 1.85, 95%CI: 1.13-3.03, P= 0.011). The findings were confirmed in multivariable analysis (HR 1.84, 95%CI: 1.11-3.07, P= 0.019). Although the frequency of the T/T variant in MMP9 rs2274755 was considerably low, the T/T variant was associated with shorter PFS and OS compared to any G allele in univariate analysis (1.7 vs. 2.1 months, HR 2.53, 95%CI: 1.17-5.46, P= 0.011; 3.9 vs. 7.6 months, HR 2.51, 95%CI: 1.11-5.72, P= 0.021) and multivariable analysis (HR 3.04, P= 0.009; HR 5.20, P< 0.001). No significant interaction was shown between the SNPs and cohorts. Disease control was significantly infrequent in patients carrying the T/T variant in MMP9 rs2274755 ( P= 0.046). Conclusions: Our data suggest that MMPs network potentially mediates tumor angiogenesis responding to regorafenib. The MMP2 and MMP9 SNPs may predict efficacy of regorafenib in patients with refractory mCRC. However, this preliminary finding warrants further validation.
Epithelial mesenchymal transition in an advanced BRAF mutation type colorectal cancer.
Authors: Hwang Jun-Eul , Bae Woo Kyun , Shim Hyun-Jeong , Cho Sang-Hee , Chung Ik-Joo
Published: January 2019

693

Background: BRAF mutation is associated with poor survival in colorectal cancer. We aimed to generate genomic signature associated with BRAF mutation that possibly predict prognosis in colorectal cancer. Methods: A gene expression signature reflecting BRAF mutation was generated in TCGA cohorts (n = 207). The colorectal cancer patients were stratified into two groups according to this signature: BRAF mutation type colorectal cancer or BRAF wild type colorectal cancer. Prognostic significance of BRAF mutation-associated gene signature was tested in three independent cohorts (GSE 17536, GSE 14333, and GSE 39582). Results: The BRAF mutation signature was associated with poor prognosis in two independent cohorts (total n = 522). BRAF mutation type colorectal cancer was associated with poor disease-free survival (median: not reached, P = 0.0303) in GSE14333, and associated with poor overall survival (BRAF mutation vs. wild, P = 0.0355), poor disease-free survival (P = 0.00794), and poor disease-specific survival (P = 0.0341) in GSE 17536. In GSE 39582, BRAF mutation type colorectal cancer demonstrated the trend of poor overall survival according to increase of stage. In a multivariate analysis, BRAF mutation gene signature was independent poor prognostic factor for disease-free survival (hazard ratio 2.7; 95% CI 1.59-2.83: P = 0.001). Gene network analyses suggested epithelial-mesenchymal transition and colon cancer metastasis signaling are the possible explanations for poor prognosis of BRAF mutation type colorectal cancer. Conclusions: BRAF mutation signature is highly associated with poor prognosis in colorectal cancer, especially in advanced stage, and the molecules associated with epithelial-mesenchymal transition can be potential therapeutic targets in BRAF mutation type colorectal cancer.
COX2 and HLA immunohistochemical (IHC) staining in colorectal tumour samples from patients participating in the ASCOLT clinical trial of adjuvant aspirin therapy.
Authors: Segelov Eva , Sieber Oliver , Chia John Whay Kuang , Williams David S
Published: January 2019

695

Background: ASCOLT is an international Phase III randomized control trial (NCT00565708) investigating the impact of adjuvant aspirin on recurrence and survival after curative resection of colorectal cancer (CRC). Retrospective data suggest that COX2 and HLA Class 1 antigen expression may predict for aspirin benefit (Reimers MS, JAMA internal medicine, 2014;174(5):732-739; and Chan AT, JAMA, 2009;302(6):649-658.) Translational studies aim to confirm putative biomarkers of aspirin sensitivity in this prospective trial. This study reports the IHC assessment of COX2 and HLA in representative samples. Methods: FFPE slides from consenting Australian patients were stained with COX2 and HLA Class 1 antibodies (Chan AT, JAMA, 2009;302(6):649-658.). Digital images were captured using Aperio software. IHC expression was scored by a pathologist (DW) blinded to sample identification. Intensity was assessed semi-quantitatively for each antibody in one area of dominant (primary) intensity and one of secondary intensity in each section. A scale was applied for staining intensity: 0=absent, 1=weak, 2=moderate and 3=strong; and for amount of tumor epithelium stained: 1= 5%-24%, 2= 25%-49%, 3=50%-74% 4= >75%. Normal epithelium, which typically has strong COX2 and HLA staining, was used as internal control. Only tumor epithelium was scored (not inflammatory cells). Results: 90 tumors were stained for both COX2 and HLA Class 1. All tumors showed diffuse staining for both markers, mostly strong, but with many tumors having regions with lesser degrees of expression (Table). Conclusions: COX2 expression was generally strong and HLA Class 1 was more likely to show regions of weak to moderate expression. No significant correlation was detected between COX2 and HLA expression levels. Further pathological analysis is underway and correlations will be made with outcome once trial data has matured in 2020. Clinical trial information: NCT00565708. [Table: see text]
Prognostic value of progranulin in patients with colorectal cancer underwent curative resection.
Authors: Koo Dong-Hoe , Do In-Gu , Oh Sukjoong , Kim Kyungeun , Sohn Jin Hee , Park Soo-Kyung , Yang Hyo-Joon , Jung Yoon Suk , Dong Il Park , Jung Kyung Uk , Kim Hyung Ook , Kim Hungdai , Serrero Ginette , Chun Ho-Kyung
Published: January 2019

696

Background: Progranulin (PGRN), characterized as an autocrine growth and survival factor, is known to stimulate the tumorigenesis and proliferation of several cancer cell types. Evidence has emerged for a potential role of PGRN as biomarkers in some diseases, however, little is known about the prognostic role of PGRN in colorectal cancers (CRCs). Methods: A retrospective analysis was performed on patients with CRCs underwent curative resection between May 2013 and June 2015. PGRN expression in tumor cells was semiquantitatively categorized (no expression, 0; weak/focal, 1+; moderate/focal or diffuse, 2+; strong/diffuse, 3+). PGRN was considered positive when either tumor cells were shown with 2+ or 3+ staining intensity. Results: A total of 109 patients (28 stage I, 32 stage II, and 49 stage III) were analyzed. The median age was 67 years (range, 30–86) and 66 patients (61%) were male. Thirty-eight patients (35%) had high tumor PGRN expression (PGRN-positive), and there was a trend of elevated pre-operative CEA and CA19–9 in patients with PGRN–positive tumor compared to those with PGRN–negative tumor (CEA, 49% vs. 21%, P = 0.003; CA19–9, 21% vs. 7%; P = 0.077). The median follow-up duration for surviving patients was 28.9 months (interquartile range, 22.5–33.9). The 3–year RFS and OS were 83.7% (95% CI, 76.8–90.6) and 96.0% (95% CI, 92.3–99.7), respectively. Patients with PGRN–positive tumors had a worse recurrence free survival (RFS) with a 66.8% at 3–year (95% CI, 51.8–81.8) compared to PGRN–negative patients with a 92.4% at 3–year (95% CI, 86.2–98.6; P = 0.01). Multivariate analysis for RFS showed that PGRN–positive tumors (hazard ratio [HR] 4.61, 95% CI, 1.26–16.93; P = 0.021) and age ( > 66 years, HR 8.87, 95% CI, 1.94–40.53; P = 0.005), stage (III, HR 8.56, 95% CI, 2.02–36.27; P = 0.004) and perineural invasion (HR 4.64, 95% CI, 1.05–20.62; P = 0.043) were prognostic factors independently associated with poor RFS after adjusting for possible confounding factors including sex, MSI status, tumor location, KRAS status, and lympho-vascular invasion. Conclusions: PGRN overexpression was significantly associated with poor RFS in patients with CRCs underwent curative resection.
Prognostic value of DNA mismatch repair status and KRAS mutations in Korean colorectal cancer patients.
Authors: Hwang Soon Woo , Cho Young-Seok , Kyeong Ok Kim , Cha Jae Myung , Joo Young-Eun
Published: January 2019

697

Background: The association of DNA mismatch repair (MMR) and KRAS mutation on clinical outcome of patients with colorectal cancer are frequently documented. However, development of more accurate prediction on clinical outcome using combination of both biomarkers remains a worthy area of investigation. The aim of this study was to evaluate the clinical relevance of KRAS mutation and MMR status in Korean patients with colorectal cancer (CRC). Methods: Clinical and pathological information for 1174 patients, who underwent surgery for colorectal cancer in 3 teaching hospitals from 2011 to 2013, were reviewed and recorded. Mutation analysis for exon 2 of KRAS gene was performed by Sanger sequencing. Expression of MMR proteins including MLH1 and MSH2 was evaluated by immunohistochemistry. Results: The overall frequencies of KRAS mutation was 34.9%. Mutations in KRAS codon 12 and codon 13 were detected in 26.7% and 8.2% of patients, respectively. Overall, 90 s (7.7%) tumors that had at least loss of expression for at least one MMR protein were defined as MMR protein deficient (MMR-D), and the remaining tumors were classified as MMR protein intact (MMR-I). According to KRAS mutation and MMR status, CRC was classified to 4 groups: Group 1, KRAS-mutated and MMR-I; Group 2, KRAS-mutated and MMR-D; Group 3, KRAS wild and MMR-I; and Group 4, KRAS wild and MMR-D. We found that patients in Group 4 had the best prognosis, and patients in Group 2 had the worst overall survival. KRAS mutation was independently associated with shorter time to recurrence and poorer overall survival. Conclusions: Combination status of KRAS mutation and MMR provide fundamental genetic signatures affecting tumor behavior, and may be used as a prognostic biomarker for CRC patient. These findings should be validated by further larger prospective study. [Table: see text]
Serological assessment of type ΧVΙ collagen in patients with colorectal cancer and ulcerative colitis.
Authors: Jensen Christina , Nielsen Signe Holm , Mortensen Joachim Høg , Kjeldsen Jens , Klinge Lone Gabriels , Krag Aleksander , Harling Henrik , Jørgensen Lars N , Karsdal Morten A , Willumsen Nicholas
Published: January 2019

698

Background: Altered extracellular matrix (ECM) remodeling is an important part of the pathology of gastrointestinal (GI) disorders. In the intestine, type XVI collagen (col-16) plays a role in pathogenesis by affecting ECM architecture and induce cell invasion. Measuring col-16 in serum may therefore have biomarker potential in GI disorders such as colorectal cancer (CRC) and ulcerative colitis (UC). The aim of this study was to determine whether col-16 can serve as a biomarker for altered ECM remodeling in patients with CRC and UC. Methods: A monoclonal antibody was raised against the C-terminal end of col-16 (C16-C) and a competitive enzyme-linked immunosorbent assay (ELISA) was developed and technically validated. Levels of C16-C were measured in serum from patients with CRC (before (n = 50) and three months after (n = 23) tumor resections), UC (n = 39) and healthy controls (n = 50). Results: The C16-C ELISA was specific towards the C-terminal of col-16. C16-C was significantly elevated both in serum from patients with CRC (p = 0.0026) and UC (p < 0.0001) compared to controls. No difference was detected in levels of C16-C between patients with CRC at baseline and three months after tumor resections (p > 0.999). Levels of C16-C identified patients with a GI disorder with a positive predictive value of 0.9 and an odds ratio of 12 (95%CI = 4.5-29.5, p < 0.0001). Conclusions: The newly developed assay detected significantly elevated levels of C16-C in serum from patients with GI disorders compared to controls suggesting its potential as a biomarker in this setting. Future studies are needed to validate these findings.
Prognostic significance of SATB1 expression in metastatic colorectal cancer: A Nordic prospective cohort study.
Authors: Siesing Christina , Sorbye Halfdan , Dragomir Anca , Pfeiffer Per , Qvortrup Camilla , Ponten Fredrik , Nodin Bjorn , Glimelius Bengt , Eberhard Jakob , Jirstrom Karin
Published: January 2019

707

Background: Special AT-rich sequence binding protein 1 (SATB1) is a transcription factor involved in global gene regulation. SATB1 expression has been associated with poor prognosis in numerous cancer forms, including colorectal cancer (CRC). The value of SATB1 expression in metastatic CRC (mCRC) has, however, not yet been described. The aim was to evaluate the expression and prognostic significance of SATB1 in mCRC. Methods: SATB1 expression was analysed by immunohistochemistry in tissue microarrays with tumors from a prospective cohort with 454 mCRC patients from three Nordic countries. Cox regression proportional hazards models were applied to assess the impact of SATB1 expression on overall survival (OS); in the entire cohort, and in strata per chemotherapy and mutational status. Results: Among the 454 evaluable cases, 144 (32%) were positive and 310 (68%) were negative for SATB1 expression. Positive SATB1 expression was significantly more frequent in cases with lung metastases (41% vs 29%, p = 0.018), but did not correlate with other sites of distant metastasis. SATB1 expression (positive vs negative) was not prognostic in the entire cohort. The prognostic value of SATB1 did not differ between patients receiving palliative treatment (n = 281) and untreated patients (n = 178), and did not differ in relation to type of chemotherapy. In patients with BRAF wild type tumors, SATB1 expression was significantly associated with prolonged OS, whereas the opposite was seen in BRAF mutated tumors (hazard ratio = 0.77 vs 1.60, p for interaction = 0.005). SATB1 expression was significantly associated with prolonged OS in patients with KRAS mutated tumors (hazard ratio = 0.68), but not in KRAS wild type tumors, and there was no significant prognostic interaction between SATB1 expression and KRAS status. The prognostic value of SATB1 expression did not differ by tumor location. Conclusions: The prognostic value of SATB1 expression in mCRC appears to depend on the mutational status of the tumors, in particular BRAF. The potential value of SATB1 expression as a predictive biomarker for targeted therapies merits further investigation.