bluuebunny/crossref_metadata_2025 huggingface dataset:
Circulating endothelial cells (CECs), progenitors (CEPs), and circulating tumor cells (CTCs) for prediction of response in patients with advanced breast cancer (ABC) receiving metronomic oral vinorelbine (oV): Preliminary results
Authors: Nole F. , Munzone E. , Bertolini F. , Sandri M. T. , Petralia G. , Adamoli L. , Radice D. , Cullura D. , di Pietro A. , Goldhirsch A.
Published: August 2020

e14572

Background: Metronomic administration of chemotherapy given once or more per week with no extended gaps was shown to be effectively anti-angiogenic, causing growth arrest or apoptosis of endothelial cells in tumor neo-vessels. Preclinical and clinical studies indicate that ultra-low concentrations of various microtubule inhibitors inhibit proliferation or migration of endothelial cells. We investigated in a phase II study the activity of metronomic administration of oV in ABC, kinetics and response prediction of CECs, CEPs, CTCs and of other biomarkers of angiogenesis (soluble VEGF, VEGFr2, TSP1, bFGF). CT perfusion scans were also performed. Methods: From February 2008, 47 pts with ABC received oV (50 mg/die TTW). Currently 20 pts are evaluable for both activity and biomarker assessment. Baseline levels of biomarkers of angiogenesis were correlated with clinical response. Results: Shown in Table . Conclusions: We found that the baseline value of apoptotic cells (expressed as % of total cells) was significantly correlated with outcome. The baseline total, viable, and apoptotic CEC count and CTCs might provide an indirect measure for angiogenic turnover and an indicator of better response to anti-angiogenic therapy, supporting the use of metronomic treatments in patients expressing high levels of baseline CECs. Updated results will be presented together with correlation with perfusion CT scan and levels of CTCs.

[Table: see text]

No significant financial relationships to disclose.
The anticancer drug development pipeline of the pharmaceutical (P) and biotech (B) industries
Authors: Thomas F. P.
Published: August 2020

e14589

Background: The rapidly growing market for anticancer drugs has attracted numerous companies, and this therapeutic area represents more that a third of all drug development candidates. We studied the pipeline of Pharma (P) and Biotech (B) companies to look at possible differences and 2008 changes in composition. Methods: The anticancer drug pipeline including candidates in Phase I to III and marketed products still actively developed in new indications was reviewed by systematically searching the websites of 41 P and 269 B active in oncology. Results: B, defined as entrepreneurial R&D-focused companies had 463 molecules including 160 in Phase I, 202 in Phase II, 73 in Phase III, and 28 marketed. P develop 219 molecules including 78 in Phase I, 61 in Phase II, 38 in Phase III, and 42 marketed. The number of candidates decreased by 10 in B and increased by 15 in P pipelines during H2, 2008, translating difficult economic conditions for B (with few candidates entering Phase I) and an increased focus of P on oncology (internal R&D efforts and B acquisition that accounts for 39 of its 219 molecules). B licensed out 119 of its candidates to P or other B. The portfolio of B and P analyzed by mechanism of action significantly differs. Symptomatic/adjuvant treatments represent 7.1 vs 5.5%, targeted small molecules 28.7 vs 42.9%, cytotoxics including reformulations 20.5 vs 22.4%, cell/gene therapies, oncolytic viruses, and oligonucleotides 7.3 vs 0%, antibodies and related proteins 19% vs 17.8%; immunotherapy 13.1% vs 5%, photodynamic and radio- therapy 2.8% vs 0.5% for B vs P, respectively. The number of antibodies and targeted small molecules increased during H1, 2008 by 2.1 and 4.4%, respectively. P recently enriched its antibody portfolio through B acquisition, but completely moved out of cytotoxics, except mid-size European and Japanese P. An update as of May,1st 2009 with a trend analysis including the potential impact of the financial crisis on B portfolio will be presented. Conclusions: The oncology pipelines of P and B are broad, different for B and P, and rapidly changing in composition.

No significant financial relationships to disclose.
Effect of metronomic use of zoledronic acid (ZOL) on antitumor and antiosteoclastic effects in breast cancer patients with bone metastasis
Authors: Xi-Chun H. , Zhao X. , Xu X. , Guo H. , Wang Z. , Guo X. , Chen J. , Wu J. , Shao Z. , Li J. , Zhu B.
Published: August 2020

e14603

Background: Zoledronic acid (ZOL) can reduce the risk of skeletal-related events (SREs) and may have direct and indirect antitumor effects, which have been shown in animal models, pilot clinical studies as well as in recent phase III randomized trials. However, the pharmacokinetics of the drug in breast cancer patients remains to be elucidated and optimized. The purpose of this randomized study was to compare the effects of ZOL on osteoclasts and angiogenesis between a weekly low-dose versus a conventional dosage. Methods: Sixty breast cancer patients with bone metastases were recruited in this randomized phase II clinical study. The participants either received ZOL 1mg IV weekly for 4 doses or a single dose of ZOL 4mg IV. No other antitumor treatments were administered. During the first month after initial infusion of ZOL, serial blood samples were collected on day 1, 15 and 29 measuring markers for bone resorption (NTx), angiogenesis (VEGF), and tumor burden (CEA and CA15–3). Results: Compared to a single-dose administration, weekly low dose of ZOL resulted in a greater reduction in serum levels of VEGF and NTx, with a significant trend over time during one month observation. There were no statistically significant differences in circulating levels of CEA and CA15–3 between the two dosing regimens. Patients who received metronomic ZOL had a longer median time to disease progression (TTP) (7.0 months, 95%CI, 6.1–7.9 months) than those who had a single dose of ZOL (2.8 months, 95%CI, 0–5.7 months; p=0.076). Conclusions: The metronomic use of low-dose ZOL 1 mg appeared to be more effective than the conventional regimen in the long-lasting reduction of VEGF and NTx, and in prolonging TTP. This dosing schedule should be further assessed in phase III trials as we demonstrated that ZOL 1mg has greater antitumor properties in our study.

No significant financial relationships to disclose.
Sorafenib and radiation: A promising combination in colorectal and pancreas cancer
Authors: Wilson G. D. , Suen A. , Robertson J. , Galoforo S. , Marples B.
Published: August 2020

e14622

Background: Sorafenib inhibits both RAF kinase, a critical component of the RAS/RAF/MEK/ERK signaling pathway that controls cell proliferation, and VEGFR-2/PDGFR-beta signaling cascade, blocking tumor angiogenesis; this dual activity has great potential for radiosensitization. This study examines the effectiveness of radiation and sorafenib in a series of human colorectal and pancreatic cell lines and xenografts with differing ras mutational status. Methods: Three colorectal cell lines were selected. HT29 and SW48 are wild-type ras whilst SW480 has a mutation in codon 12 . Two pancreatic cancer cell lines were selected, Panc 03.27 is wild-type ras whilst Panc 02.03 has a mutation in codon 12. MTT assays were used to find the best drug concentration and timing to combine with radiation. Radiosensitization was assayed using clonogenic assays with either 5 or 10μg/ml sorafenib. Xenografts were established and randomized to control, sorafenib at 20mg/kg daily (5 days/week) for 3 weeks, radiation given as 2.5 Gy daily fractions (5 days/week) for 3 weeks or a combination of the two with sorafenib administered immediately after radiation Results: The MTT data suggested that sorafenib given immediately after or at least within 2 hours of radiation appeared to give the greatest growth inhibition. Sorafenib had no effect on cell survival in the colorectal cell lines but showed a modest radiosensitization, the surviving fractions at 2 Gy alone or with 5 or 10 μg/ml sorafenib were 34%, 26% and 23% (p=0.0018) in SW48 cells. In pancreatic cells, sorafenib was toxic at 10 μg/ml and reduced survival by 30% at 5 μg/ml without sensitization The endpoint in the xenograft studies was tumor growth to 2000mm3 within a period of 120 days. In SW48 tumors, the number of days to grow to 2,000 mm3 was 13.6 ± 2.2 in the control arm, 37.0 ± 21.2 in the radiation alone arm, 17.0 ± 8.2 in the sorafenib arm and100.8 ± 28.0 in the combined arm (2 of the 5 animals were ‘cured‘). The respective data in the HT29 xenografts was13.2 ± 2.1, 62.6 ± 9.8, 38.2 ± 26.5 and106.2 ± 36.2 days (3 of 5 animals were ‘cured‘. Conclusions: Sorafenib and radiation are a promising combination. Phase I studies are under development in both pancreatic and colorectal cancer at our Instittution.

No significant financial relationships to disclose.
A comparison of potency of and intracellular signaling events provoked by, small molecule and protein-based mpl agonists
Authors: Hartley C. , Dyas G. , Archibeque I. , Al-Assaad A. , Sun B. , Lincoln P. , Zhan J. , Arvedson T. , Molineux G.
Published: August 2020

e14624

Background: Thrombopoietin (TPO) interacts with its receptor (mpl) to initiate signal transduction and increase platelet production. Other mpl agonists have been developed for clinical use: MDGF (a recombinant protein) and romiplostim (a peptibody) are protein-based mpl agonists; eltrombopag (a small molecule) is a nonpeptide mpl agonist. Methods: Mpl agonists were compared for their ability to induce cellular responses and activate relevant signaling pathways in mpl-expressing cells. Proliferation was assessed by ATP bioluminescence in Baf3/mpl cells treated with mpl agonists for 2 days. Differentiation was assessed by flow cytometric-detection of CD41 and CD61 expression on human CD34+ cells cultured with mpl agonists for 7 days. Activation of signaling pathways was determined by Western blot analysis of proteins from serum-starved Baf3/mpl cells treated with mpl agonists for 20 minutes. Experiments were repeated 3 or more times. Results: The TPO concentration that produced a half-maximal (EC50) proliferative response of Baf3/mpl cells was comparable with that of romiplostim (Table). In contrast, a 30-fold greater concentration of MGDF and a 2,000-fold greater concentration of small molecule were required to produce the same degree of proliferation. In addition, a 1,000,000-fold greater concentration of small molecule compared with romiplostim was required to produce an EC50 for differentiation of CD34+ cells. Mpl agonists activated identical signaling pathways in Baf3/mpl cells: a similar dose-response for phosphorylation of JAK2, Stat3, Stat5, Erk1/2, and AKT was observed at their respective EC10, EC50, and EC100 concentrations. Conclusions: In Baf3/mpl cells, the small molecule was between 1,000- and 1,000,000-fold less potent than protein-based mpl agonists when corrected for molecular mass differences. Small molecule and protein-based mpl agonists were indistinguishable when mpl intracellular signaling was compared at similarly effective concentrations. [Table: see text]

[Table: see text]
Design and development of high-drug-load paclitaxel (PTX)-antibody conjugates: Synthesis and preliminary evaluation of soluble paclitaxel (sPTX) and sPTX-cetuximab conjugates
Authors: Safavy A. S. , Quiles S.
Published: August 2020

e14634

Background: Tumor cell surface receptor-targeted therapy of cancer is an effective treatment strategy and monoclonal antibody (MAb)-drug conjugates hold promise as tumor-specific drug delivery systems. We have previously reported the synthesis and biological evaluation of paclitaxel (PTX)-MAb conjugates using the anti-EGFR MAb, cetuximab, as an agent for tumor-targeted drug delivery system (Safavy et al., Bioconjugate Chem 14:302–310, 2003). One limitation of these conjugates was a low drug load, i.e., PTX : MAb ratios of ≤4, and thereby, low target dose delivery. Attempts to synthesize conjugates with PTX : MAb of >4 failed due to their low water solubility. As the source of this problem was identified to be the highly lipophilic PTX moiety, we hypothesized that conjugation of water-soluble PTX analogues may lead to the preservation of the MAb solubility, and thus, to the preparation of conjugates with high PTX : MAb ratios. Methods: Newly available “discrete” poly(ethylene glycol) (dPEG) linkers were utilized for the synthesis of soluble PTX analogues. In contrast to the regular PEGs wit dispersed molecular weight ranges (MWs), the dPEGs are single-MW compounds which make possible the exact dose calculation for the resulting conjugates. Solid-phase peptide synthesis methodology was used to prepare a trifunctional dPEG-lysine (K) linker, which was first conjugated to PTX to yield the water-soluble drug analogue sPTXdPEGKdPEG, containing a free carboxylic acid group. Using the latter functionality, sPTXdPEGLysdPEG was covalently conjugated to cetuximab (C225) at increasing molar ratios. Cell binding assays were performed using MCF-7 human breast carcinoma cell line. Results: sPTXC225 conjugates with PTX : C225 ratios of up to 8 were synthesized through this method, which in contrast to the previously reported conjugates, demonstrated complete aqueous solubility. Cell binding assays indicated preservation of the immunoreactivity of the parent MAb in all conjugates. Conclusions: Water-soluble and high-drug-load PTX-MAbs such as sPTXC225, may be useful for targeted delivery of therapeutically effective doses of paclitaxel and warrant further investigation.

No significant financial relationships to disclose.
Effect of the apoptosis signal ceramide (C6) on antitumor activity of chemotherapeutic agents in SCID mice
Authors: Nadeem A. , Wanebo H. , Shrayer D. , Hazelwood S. , Resnick M.
Published: August 2020

e14642

Introduction: The ceramides are a major signaling pathway for apoptosis in cells undergoing stress or exposure to chemotherapy. We have demonstrated synergistic anti-tumor effects of combining C6 ceramide with paclitaxel, doxorubicin and cisplatin and are currently addressing the question; does C6 augment activity of all the major classes of drugs? Backround: Currently the in vivo anti-tumor effects of C6 with oxaliplatin and Gemcitabine. Methods: Invivo experiments SCID/Beige/Taconic male mice inoculated S.C. with 2X106 L3.6 pancreatic cells were treated 4 days post tumor implant with trice weekly (3x/wk) intraperitoneal (IP) injections of paclitaxel (P) 3.0 m/kg, oxaliplatin (OX) 2.5 mg/kg, cisplatin (CP) 2.5 mg/kg, Gemcitabine (Gem) 10 mg/kg with/without ceramide 10 mg/kg. Mice were observed for 6 weeks and were autopsied when near death. (All controls died by 3rd week). Maximum tumor volume, tumor weight; body weight and survival were recorded. Results: Combination with C6 ceramide augmented the tumor reduction obtained by chemotherapy alone by 57% (while preserving body weight), and increased 6 week survival from 0% (Chemotherapy alone) to 60% with combined therapy. Mean survival was increased from 25 to 37 days. Although short term immunohistochemical studies suggested enhanced apoptosis and increased caspase 3 production by ceramide combinations it may actually be independent of capase activation and mitochondrial activation. Conclusions: Combination therapy with the apoptotic signal C6 ceramide significantly enhanced the anti-tumor effects of the anti microtubule, alkylating Paclitaxel a DNA interculating antibiotic (doxorubicin) the alkylating/DNA adducting agents (cisplatin, oxaliplatin) and an anti metabolite (gemcitabine) suggesting generation of broad based apoptotic signals which interact with major cancer drug classes (tested).

No significant financial relationships to disclose.
Use of independent genome-wide assays to discover HOXA signature in colon and rectal cancers and validate a role in tumorigenesis
Authors: Pant S. , Hostetter G. , Armstrong B. , Bittner M. , Shack S. , Savage S. , Weiss G. J. , Ozer H. , Ramanathan R.
Published: August 2020

e15014

Background: Rectal cancer differs from colon cancer in terms of prognosis. Here we present genome-wide expression analysis of 79 colon and rectal tumors and gene dosage analysis of 34 archival colorectal tumors in paraffin (FFPE). Methods: Expression data were acquired ( http://expo.intgen.org/geo/home.do ) from Affymetrix experiments analyzed with GeneSpring software version 7.3 (Silicon Genetics, CA). Expression data were normalized ’per chip’ normalized to the 50th percentile of all values; and ’per gene’ normalized to the median expression level across all samples. Expression Project for Oncology (expO) samples included colon (n=50) and rectum (n=29). Differentially expressed genes were identified by parametric test for which variances were not assumed equal (Welch ANOVA). Array comparative genomic hybridization (aCGH) were performed separately on an independent set of FFPE colorectal tumors (n=34) by 244 K microarrays for CGH, Agilent Technologies, CA and with platform embedded analyses tools of CGH Analytics software and ADM-1 bioinformatics. Functional assay by knockout of HOXA9 promoter by lentiviral sh-RNAi construct was performed on colon cancer cell lines shown to have increased HOXA9 dosage and expression. Results: The analysis of expO colon and rectal cancers generated 42 genes with significant differential expression, 26 showed an increased expression of > 2 fold in colon versus rectal samples. Transcription factor family HOXA9 was the most highly expressed (3.8 fold) in colon versus rectal cancer. The aCGH data showed a low level gain of the HOXA gene to be the most frequent dosage alteration (38%). Validation by IHC and qRT-PCR showed 80% and 76% concordance, respectively. Functional assay by sh-RNAi in cell lines (compared to vehicle only) showed marked decrease in cellular viability (40 to 60%), marked morphologic change and a significantly increased apoptotic rate. Conclusion: Multiple genome-wide assays have identified HOXA9 as differentially dosed and expressed in colon and rectal tumors. Compelling functional data from sh-RNAi experiments suggests a tumorigenic role for HOXA9 in altered apoptosis.

No significant financial relationships to disclose.
Magnitude of benefit of the addition of bevacizumab (BEVA) to first-line chemotherapy (CT) for advanced/metastatic colorectal cancer (MCRC): Meta-analysis of randomized clinical trials
Authors: Vaccaro V. , Cuppone F. , Loupakis F. , Milella M. , Carlini P. , Nisticò C. , Falcone A. , Terzoli E. , Cognetti F. , Bria E.
Published: August 2020

e15021

Background: The monoclonal antibody against vascular endothelial growth factor BEVA has recently demonstrated to improve survival for MRC patients (pts). Nevertheless, the magnitude of the provided benefit in the daily practice is still controversial. In order to quantify the benefit of adding BEVA to CT for MCRC, a literature-based meta-analysis was conducted. Methods: Survival Hazard Ratios (HR) were extracted from prospective, randomized clinical trials (RCTs, either phase II/III) reports. HR and event-based relative risks (RR) with 95% confidence intervals (CI) were derived through a random-effect model. Differences in primary (progression-free- and overall-survival, PFS/OS) and secondary outcomes (overall, partial and complete response rates, ORR/PR/CR) were explored. Absolute differences (AD) and the number of patients needed to treat (NNT) were calculated. Heterogeneity test and a meta-regression analysis with clinical predictors for outcomes were conducted as well. A sensitivity analysis according to the trial phase-design was accomplished. Results: Five trials (2,728 pts), 2 phase II (313 pts) and 3 phase III (2,415 pts), were gathered. No significant interaction was found in the sensitivity analysis between phase II and III, although a trend showed a better PFS results for BEVA in phase II trials (p=0.057). At the meta-regression analysis female gender and rectal primary site were significant predictors for PFS (p=0.003, p=0.005). Toxicity analysis is ongoing. Conclusions: Although concerns regarding costs and toxicities still exist, BEVA significantly improves the outcome of untreated MCRC pts. The absolute benefit provided into an unselected population for molecular features should be considered of paramount importance for advanced disease.

[Table: see text]

No significant financial relationships to disclose.
A novel mutation in MUTYH-associated polyposis (MAP) syndrome
Authors: Fostira F. , Panopoulos C. , Papademitriou K. , Lypas G. , Lypas G. , Barbounis V. , Efremidis A. , Yannoukakos D.
Published: August 2020

e15022

Background: MUTYH-associated polyposis (MAP) syndrome is inherited as a recessive trait and is characterized by the intrinsic phenotypic feature of adenomas present at the individual's colorectum. In the case of non-surgical removal of these polyps, adenoma will transform to carcinoma. Colorectal tumours from mutation carriers display an excess of somatic mutations, specifically G:C to T:A transversions. The risk of colorectal cancer among biallelic mutations carriers seems to be close to 100% by the age of 60. Methods: Four families meeting the MAP phenotypic criteria were studied.Genomic DNA was purified from peripheral blood leukocytes following standard chloroform extraction. The complete coding sequences of the MUTYH-gene, including splice junctions, were amplified by Polymerase Chain Reaction (PCR) and electrophorized in an ABI Prism 310 Genetic Analyzer. Results: Analysis of the MUTYH gene revealed five different germline mutations, one of which novel and never characterized in respects of its pathogenicity. A mutation has been identified on exon 7 of the MUTYH gene at a homozygous state, in an individual with less than one hundred colorectal polyps at the age of forty. Another patient diagnosed with colorectal cancer at the age of 34 carried two maternally and one paternally inherited mutation, located on exon 14. A milder MAP phenotype characterized an individual, who was diagnosed with less than one hundred polyps at the age of 69. Genetic testing revealed that the patient was compound heterozygote for two missense variants located on exons 8 and 12 of the MUTYH gene. Conclusions: A novel mutation located on intron 6 of the MUTYH gene has been identified in two families. Our findings confirm previous observations highlighting the necessity for genetic testing of MUTYH in patients with MAP phenotypic features, as well as his/her extended family, in order to determine the phenotyping expression of each mutation and propose the appropriate clinical surveillance, which includes biennial colonoscopy after the age of 30. In specialized cases surgical intervention might be essential.

No significant financial relationships to disclose.
A phase I/II study of combination therapy of S-1 and irinotecan in patients with previously untreated metastatic or recurrent colorectal cancer
Authors: Choi Y. , Kim T. , Lee S. , Lee J. , Chang H. , Kim H. , Shin J. , Lee J. , Kang Y.
Published: August 2020

e15023

Background: To investigate S-1 and irinotecan (CPT-11) combination as an alternative to infusional 5- fluorouracil/leucovorin plus CPT-11, we performed a phase I/II trial to determine maximum tolerated dose (MTD), efficacy and toxicity in metastatic or recurrent colorectal cancer. In addition, we evaluated the association between genotypes of candidate genes and phenotypes. Methods: S-1 was administered orally at a dose of 70 mg/m2 (level I-III) or 80 (IV and V) from day 1 to 14. CPT-11 was given i.v. on day 1, stepping up to 175 (level I), 200 (II), 225 (III and IV) or 250 (V) mg/m2 . The treatment was repeated every 3 weeks. The association of the UGT1A1 genotypes (*6, *28, and *60) and CYP2A6 genotypes (*4, *7, and *9) with toxicities or efficacy were analyzed in patients who participated in phase II portion. Results: Twenty-three patients entered the phase I and 30 enrolled in phase II study. The MTD of S-1 and CPT-11 was considered to be 80 mg/m2 and 250 mg/m2, respectively. The dose-limiting toxicities (DLTs) were diarrhea and neutropenia. The recommended dose (RD) was determined at a S-1 dose of 80 mg/m2 and a CPT- 11 dose of 225 mg/m2. The overall response rate was 66.7% (95% CI, 48.7–84.6) at the RD level. Median time to progression was 7.6 months (95 % CI, 5.7–9.5). Median survival time was not reached. Grade3–4 neutropenia was observed in 53.4% of the patients. Grade 3–4 nonhematologic toxicities were diarrhea (16.7%) and asthenia (6.7%). The frequencies of UGT1A1*60, *28, and *6 allele were 25.8%, 10.3%, and 15.5%, respectively. Homozygous for *28 or *6 were not observed. All three double heterozygous for *28 and *6 experienced grade 3–4 neutropenia. The allele frequencies of CYP2A6*4, *7, and *9 were 15.5%, 8.6%, and 29.3%, respectively. There were no association between CYP2A6 genotypes and response rates or toxicities. Conclusions: The combination of S-1 and CPT-11 was effective and had manageable toxicities in patients with metastatic or recurrent colorectal cancer.

[Table: see text]

No significant financial relationships to disclose.
Perioperative chemotherapy with bevacizumab (BV) for liver metastases (LM) in colorectal cancer (CRC): McGill University pilot study
Authors: Bouganim N. , Kavan P. , Eid M. , Metrakos P. , Chaudhury P. , Batist G.
Published: August 2020

e15027

Background: Colorectal liver metastases treated with perioperative chemotherapy were previously shown to increase progression free survival. Given the survival benefit of bevacizumab in metastatic CRC, the aim of this study was to assess the efficacy and safety of bevacizumab based chemotherapy in the perioperative setting. Methods: In this single arm prospective pilot study, patients with resectable LM eligible to receive perioperative BV and chemotherapy were included. Kaplan Meier survival analysis was used to calculate overall survival and progression free survival. Results: A total of 60 patients were recruited, 41 male, with an average age of 55. Forty-three patients had synchronous LM. All but seven patients received pre and post-operative BV-based chemotherapy (34/60 oxaliplatin based, 22/60 CPT-11 based and 4/60 CPT-11 and oxaliplatin based). All patients underwent hepatectomy 6–8 weeks post last dose of BV. Overall response rate was 80% (48/60), 4pt with stable diseaase; 10% had a complete pathological response and 27% had no evidence of disease post hepatectomy with a median follow up of 33 months.8 patients progressed prior to surgery. Overall survival (OS) rates at 12, 24, 36 and 48 months were: 100%, 86%, 74% and 66% respectively and 5 year median survival of 55%. Progression free survival (PFS) was 14 months. Subgroup analysis of the data according to the chemotherapy pts received showed that PFS in the CPT-11 and the oxaliplatin arm were 13 and 15 months respectively. Most of the adverse events recorded were associated with the post-operative period and included wound healing (8pts), infections (2pts) and thromboemblic (6pts) disease. No sudden deaths or bowel perforations were reported. Conclusions: Bevacizumab-containing chemotherapy regimens in the peri-operative setting is effective in patients with colorectal liver metastases. Our 80% response rate and 10% complete pathological response is one of the highest reported and warrants further investigation in phase III trials.

[Table: see text]
Phase II trial of imatinib, bevacizumab and cetuximab plus modified FOLFOX-6 in advanced untreated colorectal cancer
Authors: Sobrero A. F. , Bennicelli E. , Pessino A. , Guglielmi A. , Mammoliti S. , Caprioni F. , Fornarini G. , Comandini D. , Sciallero S. , Ansaldi F.
Published: August 2020

e15030

Background: Imatinib inhibits PDGFR interfering with pericytes, the structural support to newly formed tumor blood vessels. It may thus synergize with bevacizumab. Microenvironment and tumor targeted agents along with chemotherapy could be a promising add-on approach. Methods: cetuximab 500 mg/m2, bevacizumab 5 mg/kg and modified FOLFOX-6 were given i.v. on day 1 and repeated every 2 weeks. Imatinib 400 mg/day per os was given continuously. Due to the cost and potential toxicity of the combination, the endpoint for this phase II study was very ambitious: at least 25% of complete response (medically or surgically achieved), lasting a minimum of 12 months in advanced untreated colorectal cancer patients with clearly unresectable disease. Results: Of 26 patients (16 with 1 site of disease), 17 completed the first 4 months of treatment according to the protocol, while 9 had to discontinue one biologic drug due to side effects (5 cetuximab, 3 imatinib and 1 bevacizumab). Grade 3–4 toxicity: diarrhea 12%, neutropenia 24%, skin rash 24%, hypersensitivity reactions 16%, asthenia 8%, neuro 8%. All patients were evaluable for response. Eleven responses ( 1 CR and 10 PR), 13 SD and 2 PD were observed, corresponding to 42% RR ( 95% CI = 23–61). The minimum follow up is 12 months; median PFS is 10 months. One patient among responders underwent radiofrequency ablation and 17 patients underwent surgery: 8 R-0, 3 R-1, 5 R-2 and one exploratory laparotomy. Major post surgical complications occurred in 5/17 patients. No evidence of macroscopic disease after the entire treatment plan was obtained in 13/26 patients: 12 surgical and 1 medical CR. Seven/13 were disease free at 6 months, but only 3 were still disease free at 12 months. ERCC1, ERCC2/XDP, GSTP1,TS,EGF, COX2, CYCLIN D, FCgR polymorphisms and K-RAS mutations were evaluated on all 26 patients, but no correlations were found with clinical outcome. Conclusions: The triple combination of biologics with modified FOLFOX-6 is feasible and tolerable as initial aggressive treatment. The primary endpoint of the study was not met . In fact the activity, 42% RR, was not outstanding and the high resectability rate, 69%, is misleading in the light of the short duration of the surgically induced CR.

[Table: see text]
Adjuvant chemotherapy for resected stage II and III colon cancer: Comparison of widely used two prognostic calculators
Authors: Bardia A. , Loprinzi C. , Grothey A. , Nelson G. , Alberts S. , Patiyil S. , Thome S. , Gill S. , Sargent D.
Published: August 2020

e15031

Background: Two web based prognostic calculators, Adjuvant! and Numeracy (JCO 22:1797–1806, 2004) are widely used to provide individual patient outcome predictions to aid decisions regarding adjuvant therapy for colon cancer. The comparability of these tools has not been studied yet. The aim of this project was to assess whether the Numeracy and Adjuvant! Colon cancer prognostic tools produced similar results for patients with resected stage II and III colon cancer based on a set of hypothetical patients. Methods: All possible hypothetical scenarios were formulated for the Numeracy calculator based on all potential combinations of age, lymph nodes status, tumor stage, and grade of tumor. These were applied to three post- surgical therapy choices: observation, 5-FU (5-fluorouracil), or FOLFOX (5-FU, leucovorin and oxaliplatin). Predictions for these hypothetical scenarios (N= 192 for males and 192 for females) were also entered into the Adjuvant! Program. Predicted relapse free survival (RFS) and overall survival (OS) were obtained using both calculators. Wilcoxon signed rank tests were used to compare the numerical predictions between the calculators for each outcome. Results: In the majority of the 384 hypothetical patient scenarios, predictions for RFS and OS from Adjuvant! were statistically significantly higher than from Numeracy (p value < 0.05), except for age≥70 for FOLFOX, and the number of positive nodes of 0 and 1–4 using FOLFOX, among males. The net estimate of benefit for RFS and OS for 5-FU over surgery, obtained from Adjuvant! and Numeracy, were similar (for both males and females), but the benefit in RFS and OS for FOLFOX over 5-FU obtained from Adjuvant! was significantly lower than the estimate obtained from Numeracy (p value < 0.05). Conclusions: Based on a hypothetical set of patients with resected stage II and III colon cancer, the estimated benefit in RFS and OS of FOLFOX over 5-FU based chemotherapy is lower in Adjuvant! than in the Numeracy tool (but benefit for 5-FU over surgery alone is similar). Further studies are needed to clarify the discrepancy and to assess which of these tools most accurately reflects actual patient outcome.

No significant financial relationships to disclose.
Prognostic significance of VDR, DKK-1, and DKK-4 expression in colorectal cancer
Authors: Posey J. A. , Koya S. , Katkoori V. R. , Davis D. E. , Manne U.
Published: August 2020

e15047

Introduction: Inactivation of the vitamin D receptor (VDR) and up-regulation of its down-stream molecules, Dickkopf-1 (DKK-1) and DKK-4 are implicated in the aggressive progression of colorectal adenocarcinomas (CRC). In this study, the expression of these molecules was evaluated in sporadic CRCs, and their expression levels were assessed for correlation with patient prognosis. Methods: By the qRT-PCR, 98 CRCs and matching normal tissues were analyzed for expression of VDR, DKK-1, and DKK-4 at the mRNA level. These levels were normalized to b-actin expression and calculated as ratios of copy numbers. The expression levels were assessed for correlation with clinicopathological features and for their prognostic importance. The survival probabilities were estimated by the Kaplan-Meier method. Results: VDR expression was 1.5 times lower in CRCs (53 of 98, 54%) than in their corresponding normal tissues. Expression of DKK-1 was found in 51 of 98 (52%) of the CRCs and in 29 of 98 (29%) of the normal tissues. Expression of DKK-4 was found in only 14 of 98 (14%) of the CRCs and in none of the normal tissues. VDR expression was 2.0 fold lower in tumors that exhibited metastasis, and poor differentiation and in large tumors ( > 5) cm as compared to tumors without metastasis, well or moderately differentiated tumors and tumors of small size (< 5 cm), respectively. In contrast, increased (3-fold) expression of DKK-1 and DKK-4 was observed in tumors with distant and/or nodal metastases as compared to tumors without such metastases. Expression of DKK-1 was higher in tumors with poor differentiation and in large tumors. There was an inverse relationship between VDR and DKK-4 expression. Survival analyses suggested that CRCs expressing DKK-1 (log rank, P = 0.046) and those with decreased or lack of expression of VDR (log rank, P = 0.043) were associated with poor patient survival. There was, however, no prognostic value for DKK-4 expression (log rank P=0.624). Conclusions: These findings suggest that, for CRCs, increased expression of DKK-1 and reduced or lack of expression of VDR, are associated with aggressiveness and with a poor prognosis for patients. These results provide evidence augmenting the VDR-DKK- pathway may represent a rational therapeutic strategy in colorectal carcinoma.

No significant financial relationships to disclose.
Development of an assay for KRAS mutation as a predictive marker for cetuximab in colorectal cancer
Authors: Yokota T. , Muro K. , Shibata N. , Ura T. , Takahari D. , Shitara K. , Yatabe Y.
Published: August 2020

e15048

Background: The activating KRAS mutation is an important predictive marker for anti-EGFR therapy against colorectal cancer (CRC), although valid assays using pathological specimens have not been yet established. We present a rapid, sensitive assay for KRAS genotyping using small biopsy specimens. Methods: We used Cycleave PCR to detect the KRAS mutation in CRC with a chimeric DNA-RNA-DNA probe labeled with fluorescent dye and quencher, with results obtained within 4 hours. Template DNA was extracted from formalin-fixed, paraffin-embedded specimens, which are surgically resected or biopsied in clinical practice. Results: To evaluate Cycleave PCR accuracy for detecting the KRAS mutation, we compared with results of reverse transcriptase-PCR-coupled direct sequencing (RT-PCR-DS) of metastatic lung tumor specimens from CRC patients. KRAS mutations were present in 40 (43%) of 94 patients, including 28 (30%) and 8 (9%) in codon 12 and codon 13 mutations, respectively. Concordant results between Cycleave PCR and RT-PCR-DS in KRAS codon 12 and 13 were found in 35/36 (97%) and 23/23 (100%), respectively. We also applied this method to surgical specimens in clinical practice. Although 8 from 73 patients (11%) could not be evaluated with Cycleave PCR, corresponding biopsy specimens could be used alternatively. Because biopsy specimens were fixed by formalin for a shorter period, fixation of surgical specimens for longer time may have contributed to PCR failure. Indeed, over- fixation by formalin impaired PCR amplification of KRAS in time-dependently. Furthermore, results of 4 randomly selected biopsy specimens using Cycleave PCR were consistent with those of surgical specimens. Conclusions: Cycleave PCR even using biopsy specimens is accurate, rapid, and useful to detect KRAS mutations in CRC patients. This study also called attention to specimen selection, as over-fixation by formalin may lead to failure of the gene examination due to template DNA fragmentation. This method can be applied to examine BRAF genotyping as well as KRAS. KRAS/BRAF genotyping by Cycleave PCR leads to individualized therapy using cetuximab for CRC patients.

No significant financial relationships to disclose.
High- versus low-dose leucovorin in 5-fluorouracil-based oxaliplatin- or irinotecan-containing regimen in metastatic colorectal cancer
Authors: Song E. , Lee N. , Kwak J. , Yhim H. , Lee K. , Roh H. , Sohn M. , Yim C.
Published: August 2020

e15054

Background: Although leucovorin (LV) is widely used as a modulator of 5-fluorouracil (5-FU) for the chemotherapy of advanced colorectal cancer, the optimal dose has not yet been established. Low-dose LV appears to be as active as high-dose LV in the several studies. So, we tried to compare the efficacy of high-versus low-dose LV in the commonly used palliative chemotherapy regimen. Methods: Between May 2003 and May 2008, 40 patients with metastatic colorectal cancer were randomly treated with high-LV (200mg/m2/i.v.) or low-LV (20mg/m2/i.v.) in 5-FU based oxaliplatin (FOLFOX-4) or irinotecan (FOLFIRI) containing regimen. The primary endpoint of the study was the comparison of response rates and the secondary endpoint was the assessment of survival and tolerability. Results: The response rate was 40% in low-LV group with 2 CR and 6 PR, and 35% in high-LV group with 2 CR and 5 PR, without any significant difference (P = 0.89). The median overall survival was 24.3 months in low-LV group and 25.2 months in high-LV group, with no difference between treatments. Toxicity mainly consisted of gastrointestinal side effects, which were rare and similar in the two groups. Conclusions: In this randomized phase II study, the low and high doses of LV appeared to be equivalent in palliative chemotherapy of metastatic colorectal cancer although large-scale phase III study are necessary.

No significant financial relationships to disclose.
Bevacizumab (BV) plus chemotherapy (CT) as first-line treatment of patients with metastatic colorectal cancer (mCRC): First results from a large community-based observational cohort study in Germany
Authors: Arnold D. , Kindler M. , Petersen V. , Tummes D. , Moelle M. , Schoeberl C. , Srock S. , Kutscheidt A. , Grothe W.
Published: August 2020

e15057

Background: BV prolongs overall survival (OS) and progression-free survival (PFS) when added to standard CT for patients (pts) with mCRC. After approval of BV in Germany in 01/05, this observational cohort study was initiated in pts receiving BV with various first-line CT regimens to evaluate safety events and effectiveness. Methods: To facilitate and evaluate enrollment of a typical mCRC population, eligibility criteria were minimized. Choice of the CT regimen was at physician´s discretion, but influenced by the current registration status. Predefined endpoints were treatment characteristics, response rate (RR), PFS, OS, and adverse events assessed as potentially related to the treatment (AERT) or as severe (SAE). Pts are followed for up to 4 years, and clinical data were updated every cycle (2–4 weeks). Results: 1,300 pts were enrolled at 261 sites from 01/05 through 06/08; abstract data cutoff date was 11/24/08. Median age: 64 (range 19–100) years, > 70 years.: 25%; male 63%; ECOG PS status 0–1/2/>2 88%/11%/1%. CT choice was fluoropyrimidine (FU)/BV alone (any 5-FU or capecitabine) 12%; FU/BV/oxaliplatin 18%; FU/BV/irinotecan 69%; other 1%. Median treatment duration with BV was 7.4 (range 0–28) months; duration > 10 months 21%. 60-day mortality was 4.0%. In total, 183 AERT were reported in 10% of pts., most commonly diarrhea (34 AERT), nausea (26) and hypertension (18). Reported SAE were deep venous TE in 0.8% of pts, bleeding 1.2%, pulmonary embolism 0.5%, arterial TE 0.4%. Best investigator- assessed RR was 59% (CR 10%, PR 49%; all pts) with 64% for FU/BV/oxaliplatin, 59% for FU/BV/irinotecan and 53% for FU/BV alone. Conclusions: The safety profile of BV in this population of mCRC pts with different CT regimens appears consistent with that observed in the other reports like BRiTE or first BEATrial. RR in this preliminary dataset are higher than that reported in the randomized controlled trials, but likely biased by the methodology of evaluation. However, activity with FU/BV alone was surprisingly high. Updated efficacy data including PFS will be presented.

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Targeted plasma proteome profiling for early prediction of chemotherapy response and toxicity in colorectal cancer (CRC)
Authors: Chua W. , Randall S. , McKay M. J. , Horvath L. , Clarke S. J. , Molloy M. P.
Published: August 2020

e15063

Background: Accurate predictors of chemotherapy response and toxicity in CRC are required to improve the safety, efficacy and costs of treatment. Plasma proteomic profiling using multiple reaction monitoring mass spectrometry (MRM-MS) couples assay multiplexing with high specificity to determine levels of pre-selected biomarker proteins. Our aims were to investigate the utility of plasma proteomic profiling using MRM-MS for predicting: (1) early haematological toxicity, (2) response and (3) survival for patients receiving chemotherapy for CRC. Methods: Patients with locally advanced and metastatic CRC receiving chemotherapy were enrolled. Plasma collection was performed pre-chemotherapy, at day 3 and day 15 of treatment. Toxicity assessments (NCI Criteria v3.0) were prospectively collected for all patients and treatment response (RECIST) assessed for patients with metastatic disease. MRM-MS assays were designed for 39 peptides representing 31 liver derived plasma proteins with reported roles in inflammation and/or cancer. Statistical analysis was performed using the 2-sample t-test to assess statistically significant fold change differences (p<0.05) between sample days for: (1) patients with absence or presence of ≥ Grade 2 neutropenia after two cycles and (2) responders (CR and PR) versus non-responders (SD or PD). Results: Plasma proteomic profiling for 39 peptides was performed for 17 patients. The greatest change in protein levels was observed between Day 3 and 15 with 9% of proteins showing a 1.5 fold or greater change with some proteins showing a ≥ 200-fold change in level. Three proteins (clusterin, paraoxanse and apolipoprotein A1) were significantly different in the group analysed for neutropenia and four proteins (kininogen I, serum paraoxonase/ arylestease 1, apolipoprotein A-1 and complement C4a) were significantly different in the group analysed according to response. Survival analysis was not performed due to the short follow up. Discussion: Our results are encouraging for the use of a robust mass spectrometry technique for early prediction of neutropenia and chemotherapeutic response in CRC.

No significant financial relationships to disclose.
Evaluation of thymidylate synthase and ERCC1 mRNA levels as predictive markers in colorectal cancer patients treated with S-1 and oxaliplatin
Authors: Kuramochi H. , Hayashi K. , Nakajima G. , Kamikozuru H. , Yamamoto M.
Published: August 2020

e15071

Background: Oxaliplatin has been widely used for the treatment of colorectal cancer. The mechanism of action of platinum compounds such as oxaliplatin is to bind to a DNA molecule in the form of a platinum-DNA-adduct. Excision repair cross complementation group 1 (ERCC1), which plays a major role in the nucleotide excision pathway, has a polymorphism in codon 118, and is reported to be associated with a resistance to platinum-based therapy. Thymidylate synthase (TS) and dehydropyrimidine dehydrogenase (DPD) are key enzymes of 5-FU metabolism and are well known to be associated with a response to 5-FU-based therapy. Methods: Twenty-one colorectal cancer patients (male:female = 7:14; median age, 65) treated with a combination of oxaliplatin and S-1 as a first-line therapy were analyzed for ERCC1 codon 118 polymorphism and the mRNA expression levels of TS, ERCC1, and DPD. Formalin-fixed paraffin- embedded surgical specimens were used and t-RNA and DNA were extracted. The mRNA expression levels were measured using real-time RT-PCR, and the polymorphism was analyzed using the allelic discrimination method together with real-time PCR. Results: No correlation was observed between ERCC1 codon118 polymorphism and any response to the chemotherapy. ERCC1 mRNA levels tended to be higher in the patients with wild-type homozygous alleles in codon 118 than in those with at least one mutant allele(1.19 vs.0.68: p= 0.15). Patients with both high TS and ERCC1 mRNA levels showed a significantly lower response rate than the others (25% vs. 67%, p=0.02). No relationship was seen between DPD mRNA expression levels and the response. Conclusions: The mRNA expression levels of TS and ERCC1 appear to be useful markers for the treatment of S-1 and oxaliplatin. No particular usefulness of ERCC1 codon 118 polymorphism was verified.

No significant financial relationships to disclose.