bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical significance of mutational status in gastrointestinal stromal tumors (GIST).
Authors: Arkhiri Peter , Stilidi Ivan S. , Poddubnaya I. , Nikulin Maxim P. , Tsymzhitova Natalia , Mazurenko Natalia N.
Published: March 2020

e20501

Background: Mutational status of GIST patients plays an important role in clinical practice. In most cases, mutations are localized in KIT exon 11. KIT exon 9 mutations are less common and are associated with poor prognosis. The study aims to analyze the prognostic role of mutational status and type of mutation in GIST patients. Methods: Analysis of the mutational status of 52 GIST patients with localized disease was conducted between January 1, 2001 and November 1, 2007. KIT/PDGFRA tyrosine kinase inhibitors were used in the treatment of GIST patients with diseases progression only. Results: 89.6% of KIT mutations were associated with spindle cell stromal tumors while 85.7% of PDGFRA - with gastric epithelioid GISTs. KIT mutations were discovered in 40 patients (76.9%), of which 34 cases (65.5%) exhibited 11 exon, 5 (9.6%) – 9 exon and 1(1.9%) case - 13 exon. PDGFRA mutations have been detected in 7 (13.5%) patients and all of them involved 18 exon. Deletions were detected in 17 cases: 10 in patients with gastric and 7 with small intestinal GISTs. Less frequent were substitutions, in 14 patients and duplications, in 3 patients. All 5 cases with KIT 9 exon mutations were duplications in small intestine stromal tumors. Duplications and substitutions in 11 exon KIT were associated with better clinical outcome (5-year survival rate of 100.0% and 75.0 ± 21.7%) when compared with deletions and duplications in 9 exon (5-year survival rate of 48.4 ± 14.1% and 25.0 ± 21.7%). Survival analyses with deletions KIT 11 exon showed no significant difference, with 5 year survival rate in patients with gastric stromal tumors of 47.6 ± 17.1%, compared with 53.3 ± 17.3% in intestinal tumors (g = 0.6496). Conclusions: The better prognosis was observed for patients with duplications and substitutions in 11 exon KIT and worse outcome—in patients with deletions in 11 and duplications in 9 exon KIT. No prognostic difference has been detected in patients with gastric and small intestinal stromal tumors with similar mutations (deletions in KIT exon 11). The poor prognosis in small intestinal stromal tumors patients is probably due to duplications in KIT exon 9.
Analysis of Hohn (HC) and porth-a-cath (PAC) central venous catheters (CVC) adverse events (AEs) and related costs in advanced soft tissue sarcomas (STS) treated with trabectedin (TR) 24-hour (24-h) infusion therapy.
Authors: Palesandro Erica , Boccone Paola , D'Ambrosio Lorenzo , Galizia Danilo , Aliberti Sandra , Debernardi Felicino , Bona Francesco , Manca Antonio , Anselmetti Giovanni Carlo , Russo Filippo , Campanella Delia , Aglietta Massimo , Grignani Giovanni
Published: March 2020

e20512

Background: TR is registered as second line in STS. The 24-h infusion schedule showed a better activity profile but a CVC is mandatory to administer TR with related costs and possible complications. The HC (Bard Access System, USA) is a silicone, open-ended, non-tunneled CVC characterized by low cost and easy placement and removal. We performed a retrospective analysis of STS patients (pts) treated with TR through a CVC to evaluate the incidence of CVC AEs (malfunctioning, thrombosis, infection or extravasation) in pts carrying HC or PAC. Methods: Eligible criteria were: at least 1 course of TR via CVC, adequate premedication (steroid and 4000 units/die of low molecular weight heparin), absence of previous CVC alterations or any underlying coagulation disorder. All CVC were placed in our Institution under radiological control and full medical history was available for all pts included into the study. Average total costs (placement, hospital admission, device, and medications) are 255 and 560 $ for HC and PAC, respectively. Results: Between March 2008 and October 2011 42 pts, median age 58 (20-74) yrs, received TR after placement of a CVC: 14 (33%) HC and 28 (67%) PAC. At the time of TR administration, pts had received a median of 2 (range 1-4) chemotherapeutic lines. A total of 185 courses of TR (44 via HC / 141 via PAC) were administered at a median dose of 1.3 mg/m2 with a median of 3 (1-22) courses per pts. AEs occurred after a median of 2 (range 1-6) TR courses. We recorded 13 CVC-related AEs: 7 infection (2 HC / 5 PAC), 4 thrombosis (2 HC / 2 PAC), 2 malfunctioning (1 HC / 1 PAC,) no extravasation. CVC had to be removed and substituted in 10 pts (5 HC/5 PAC) with an estimated total cost of 280 and 680 $ for HC and PAC, respectively. In PAC pts, AEs determined 5 dose delays, only. Conclusions: CVC AEs should be considered in the management of pts treated with TR. HC showed a favorable safety profile and its AEs management is easier compared to implanted CVC. If disease does not progress after 2 cycles, HC can be converted in an implanted CVC. However, HC is also an adequate and cheaper device when best supportive care is the only useful therapy left.
MicroRNAs and response to anti-EGFR therapy in metastatic colorectal cancer.
Authors: Blanco-Calvo Moisés , Haz-Conde Mar , Rodríguez-Rigueiro Teresa , Figueroa Angelica , Reboredo Margarita , Medina Villaamil Vanessa , Lorenzo-Patiño Maria J , Arnal Francisco , Antón-Aparicio Luis , Alonso-Jaudenes Curbera Guillermo , Valladares-Ayerbes Manuel
Published: March 2020

e21002

Background: The discovery of KRAS gene as response predictive factor to anti-EGFR antibodies supposed a revolution in the pharmacological management of metastatic colorectal cancer (mCRC) patients (pts), since only KRAS native (WT) pts are possible responders (R) and susceptible to be treated with this targeted therapy. However, the appearance of non-responders (NR) among KRAS WT pts during the administration of anti-EGFR therapy has stimulated the research in additional predictive markers. Here, we explore the putative role of microRNA (miRNA) deregulation as predictive tool of response to anti-EGFR therapy in mCRC pts. Methods: Tissue samples of mCRC were obtained as formalin-fixed paraffin-embedded (FFPE) specimens from the archives of our institution, after obtaining informed consent of pts. Genomic DNA was extracted using Qiaamp DNA mini kit (Qiagen). For the mutational testing of KRAS gene, we use the TheraScreen KRAS PCR kit (DxS, Qiagen). miRNA-containing total RNA was extracted using Tripure (Roche). miRNA expression was analyzed by SYBR-green-based RT-qPCR in a LightCycler 480 (Roche) device using the Human Cancer RT2 miRNA PCR array (SABiosciences, Qiagen). First data analysis was performed with the application provided by supplier (SABiosciences, Qiagen). Bioinformatic analyses were performed in miRBase 18, TargetScanHuman 6.0 and TarBase 6.0 databases. Additional statistical analyses were performed on R 2.14 (R Foundation), SPSS 19 (IBM) and Prism (GraphPad). Results: We analyze 9 primary colon cancer and 3 non-matched normal adjacent (NAT) tissues. Tumor samples corresponded to 6 KRAS WT and 3 KRAS mutated pts. Among KRAS WT pts, 3 had shown response to anti-EGFR antibodies and 3 did not. Samples were analyzed in duplicate for the expression of 88 cancer-related miRNAs. We found 22 miRNAs significantly (p<0.01) upregulated (>4-fold) when comparing KRAS WT NR versus R pts. From these 22 miRNAs, we detected only 6 when we search in databases for experimentally tested or predicted miRNA interactions with KRAS. Conclusions: We found a miRNA signature which could be utilized as response predictive tool for anti-EGFR therapy in mCRC pts. Further analysis will be necessary to establish the performance of this signature.
MYC gene copy number (GCN) and sensitivity to anti-EGFR monoclonal antibodies in metastatic colorectal cancer (mCRC).
Authors: D'Incecco Armida , Landi Lorenza , Fountzilas George , Kalogeras Konstantine T. , Geva Ravit , Kako Severine , Robb Caroline M. , Frattini Milo , Rossi Elisa , Salvini Jessica , Ludovini Vienna , Crinò Lucio , Cappuzzo Federico , Varella-Garcia Marileila
Published: March 2020

e21018

Background: Monoclonal antibodies against Epidermal Growth Factor Receptor (EGFR) demonstrated efficacy in mCRC patients without mutations in the KRAS gene. Previous data in breast and lung cancer suggested that MYC GCN affects sensitivity to anti-EGFR agents. Methods: This retrospective study was conducted in a cohort of 303 mCRC patients treated with cetuximab/panitumumab, either alone (N=24) or in combination with chemotherapy (N=279). MYC GCN was assessed by fluorescence in situ hybridization (FISH) in primary colorectal cancer tissue samples. Results: In the study population response rate (RR) was 28%, median progression-free survival (PFS) 5.6 months and median overall survival (OS) 11.8 months. MYC was successfully evaluated in 298 cases and was amplified in 17 patients (5.7%). Individuals with MYC amplification showed a trend for a lower RR (7.7% versus 28.9%, p=0.12), shorter PFS (3.0 months versus 5.8 months, p=0.22) and shorter OS (11.3 months versus 12.6 months, p=0.15) than patients with non-amplified tumors. A Receiver Operating Characteristic (ROC) analysis was also performed in order to verify whether a better MYC GCN cut-off could discriminate between anti-EGFR sensitive and resistant cases. In this analysis, patients with mean MYC GCN ≥2.53 (N=199, 66.8%) had a significantly higher RR (32.2% versus 19.1%, p=0.02), a longer PFS (5.8 months versus 4.5 months, p=0.06) and a longer OS (12.3 months versus 11.4 months, p=0.59) than patients with mean GCN <2.53 (N=99, 33.2%). In order to assess the potential confounding effect of KRAS and BRAF status, we further analyzed the outcome of the 81 KRAS/BRAF wild-type patients according to MYC GCN. In this subset, a trend for improved RR (42% versus 20%, p=0.05), PFS (8.6 months versus 4.2 months, p=0.16) and OS (13.5 months versus 9.7 months, p=0.51) favored MYC FISH GCN ≥2.73 patients. Conclusions: MYC amplification is a rare event in mCRC, not significantly reducing sensitivity to anti-EGFR agents. A trend for better outcome was observed in presence of increased MYC GCN.
Circulating tumor cells (CTCs) and molecular subtypes (MST) in a treatment algorithm for metastatic breast cancer (MBC).
Authors: Graham Mark , Fleming Natanya , Blumenthal Susan , Boehlke Brittani , Schuur Eric
Published: March 2020

e21022

Background: We measured CTC s and MST of MBC [ER+, HER2- (HR+); HER2+ (H2+); or Triple Negative (TN)] in all MBC pts in our practice and used these in an algorithm to treat MBC. Methods: CTC’s were measured in 156 pts whether beginning Rx with 1st MBC (97), or under way with Rx (59). Pts had CTC s measured over 61 mo. from 11/20/06 thr. 12/31/11; 66 of the pts had died as of 12/31/11. Pts were Rx’d with an algorithm using CTC’s and MST: Pts with ≥ 5 CTC s were Rx’d with chemoRx, usually doublet ChemoRx, but accounting for pt preferences. When CTC’s improved to < 5 for a sustained 3-4 mo. period, or if initially < 5m we sought less morbid therapies: hormonal if ER+, or single agent ChemoRx +/- biologics if ER- or hormone refractory. In a detailed statistical analysis of variables affecting Disease Specific Survival (DSS) in the first 140 pts thr. 6/30/11, with median f/u of 24.0 mo., highest CTC level (max CTC) observed during f/u was used for analysis: CTC Low (max <5, n=60); CTC Moderate (max 5-99; n=55); and CTC High (max ≥ 100; n=25). Results: We reported the distribution of pts and death rates for the 3 MST and the 3 max CTC groupings earlier (Graham, ASCO 2010); the conclusions are similar with 156 pts and longer f/u. Each MST has 40% of pts with 0-4 CTC’s at all time points. Each MST has a smaller group with CTC’s ≥ 100, 16% of all pts. 33% of deaths (22 of 66) occurred in the ≥ 100 CTC pts. 88% with ≥ 100 CTC’s died; these are early deaths. Only 13 of 66 (20%) died if CTC’s < 5, and 31 of 65 (48%) ifh 5-99 CTC’s. In the first 140 pt analyzed thr. 6/30/11, the median DSS from date of highest CTC is not reached for the CTC Low (max. f/u 54 mo.), vs. 35.8 mo. in the CTC Moderate and 3.3 mo. in the CTC High. DSS was signif. longer for the CTC Low and CTC Moderate vs. the CTC High (P<0.001 for both comparisons). DSS was also signif. greater in the CTC Low vs. the CTC Moderate (P = 0.04). Cox multivariate analysis showed that the Max CTC group (HR 4.8, 95% CI 3.1 - 7.6, p < 0.001), age ≥ 55 at time of max CTC (HR 4.0, 95% CI 2.7 - 7.9, p < 0.001), and MST (HR 2.1, 95% CI 1.4 – 3.1, p < 0.001) were predictive of DSS. Conclusions: a. A treatment algorithm of CTC’s and MST in MBC finds subgroups of MBC with long, intermediate and very short DSS; b. Effective treatments are short-lived in pts with CTC max ≥ 100.
Variations of T cells following metronomic and standard chemotherapy in patients with solid tumors.
Authors: Koumarianou Anna , Christodoulou Marianna , Papadopoulos Iordanis , Stergiopoulos Spyros , Peros George , Dimitriadis George , Tountas Nikolaos , Economopoulos Theofanis , Pectasides Dimitrios G. , Pappa Vassiliki , Xiros Nikolaos
Published: March 2020

e21027

Background: Evidence suggests that the interaction of chemotherapy with effector (Teffs) and suppressor (Tregs) T-cells may influence the survival of patients. We investigated the impact of metronomic (p.o.) and standard intravenous (i.v.) chemotherapy on Tregs and Teffs. Methods: Blood samples were collected from 37 cancer patients, before (pre) and after (post) the first cycle of i.v. (n=18) or p.o. (n=19) chemotherapy. CD4+CD25- Teffs and CD4+CD25+ Tregs were freshly isolated with magnetic-bead sorting (Miltenyi Biotec). Teffs:Tregs co-cultures were developed in the presence of CD2, CD3 and CD28 antibodies for the study of the Tregs over Teffs suppressive capacity. The cell proliferation was evaluated utilizing a colorimetric ELISA (O.D. at 450 nm) (Roche Applied Science). Statistical analyses using Mann-Whitney and Tukey’s (one-way ANOVA) tests were performed in order to determine if changes in the numerical and/or functional T-cell characteristics occurred in association with a particular chemotherapy approach. Results: Decreased numbers of total T-cells were observed in all patients regardless chemotherapy type. The post-chemotherapy group displayed higher ratio of Tregs to Teffs cells as well as higher Tregs over Teffs suppression (expressed as the % difference in Teff proliferation rate between single Teffs cultures and Teffs:Tregs co-cultures) compared to the pre-chemotherapy group (0.17±0.02 vs 0.09±0.07, p=0.002 and 30.65%±12.12 vs 10.65%±16.80, respectively). Subgroup analyses revealed that these parameters were significantly higher in patients treated with p.o. chemotherapy compared to those treated with i.v. chemotherapy (0.21±0.04 vs 0.12±0.03, p=0.01 and 44.43%±7.34 vs 16.87%±22.99, respectively). Patients receiving drugs targeting the mitotic spindle, such as docetaxel or vinorelbine, exhibited a stronger Tregs suppression (44.32%±6.78) as compared to those treated with DNA sequence-interfering drugs, such as epirubicin or cisplatin (10.30%±38.85). Conclusions: Chemotherapy influences the immune response in favor of Tregs and against Teffs in a drug-specific manner. Compared to i.v., p.o. chemotherapy correlates with greater repression of Teffs function.
Effect of targeting of syndecan-1 by microRNA miR-10b on breast cancer cell motility and invasiveness via a rho-GTPase- and E-cadherin-dependent mechanism.
Authors: Kiesel Ludwig , Ibrahim Sherif Abdelaziz , Yip George W , Gotte Martin
Published: March 2020

e21041

Background: microRNAs are small endogenous non-coding RNAs, which posttranscriptionally regulate gene expression. In breast cancer, overexpression of the transmembrane heparan sulfate proteoglycan syndecan-1, a predicted target of the oncomiR miR-10b, correlates with poor clinical outcome. Here, we investigatet the potential functional relationship of miR-10b and syndecan-1 in an in vitro study. Methods: MDA-MB-231 and MCF-7 breast cancer cells were transiently transfected with pre-miR-10b, syndecan-1 siRNA or control reagents, respectively. Altered cell behaviour was monitored by proliferation, migration and invasion chamber assays, and time-lapse video microscopy. Results: miR-10b overexpression induced posttranscriptional downregulation of syndecan-1, as demonstrated by qPCR, flow cytometry, and 3’UTR luciferase assays, resulting in increased cancer cell migration and matrigel invasiveness. Syndecan-1 silencing generated a copy of this phenotype. Adhesion to fibronectin and laminin and basal cell proliferation were increased. Syndecan-1 co-immunoprecipitated with focal adhesion kinase, which showed increased activation upon syndecan-1 depletion. Affymetrix screening and confirmatory qPCR and Western blotting analysis of syndecan-1 deficient cells revealed upregulation of ATF-2, COX- 2, cadherin-11, vinculin, ACTG2, MYL9, transgelin-1, RhoA/C, MMP2 and heparanase, and downregulation of AML1/RUNX1, E-cadherin, CLDN1, p21WAF/CIP, Cdk6, TLR-4, PAI1/2, Collagen1alpha1, JHDM1D, Mpp4, MMP9, matrilin-2 and ANXA3/A10. Video microscopy demonstrated massively increased Rho kinase-dependent motility of syndecan-1-depleted cells, which displayed increased filopodia formation. Conclusions: We conclude that syndecan-1 is a novel target of the oncomiR miR-10b. Rho-GTPase dependent modulation of cytoskeletal function and downregulation of E-cadherin expression are identified as relevant effectors of the miR-10b-syndecan-1 axis, which emerges as a promising target for the development of new therapeutic approaches for breast cancer.
Antioxidants and oxidative stress in patients with Li-Fraumeni-like syndrome and TP53 p.R337H mutation carriers.
Authors: Macedo Gabriel de Souza , Giacomazzi Juliana , Netto Cristina , Motta Leonardo , Klamt Fábio , Ashton-Prolla Patricia
Published: March 2020

e21042

Background: Germline mutations in the TP53 gene are the underlying genetic defect of Li-Fraumeni Syndrome (LFS) and its variant, Li-Fraumeni-like (LFL) Syndrome, autosomal dominant disorders characterized by predisposition to multiple early-onset cancers. More recently, p53 is emerging as an important player in redox metabolism and important enzymes involved in defenses against oxidative stress have shown to be regulated by p53, including glutathione peroxidase 1 (GPX1) and mitochondrial superoxide dismutase 2 (SOD2). The aim of the present study was to investigate the redox profile parameters in blood of p.R337H mutation carriers and non-carriers individuals. Methods: A total of 34 individuals were included in the study and they were divided in two groups: mutation carriers (n=17) and non-carriers (n=17). Antioxidant enzyme activities (SOD, CAT, GPx) and oxidative stress parameters (Protein carbonyl content, Sulfhydryl content and TBARS) were measured in plasma, erythrocytes, leukocytes and serum. Results: Erythrocyte SOD and GPx activities, two first-line players in enzimatic antioxidant defense, differed significantly between mutation carriers and non-carriers, with an increased activity in the latter (p>0,05). Plasma Carbonyl content, an indicative of protein damage related to ROS overgeneration, was also increased in carriers (p=0,015). There was no significant difference between groups in all other parameters evaluated. Conclusions: Our findings suggest that TP53 p.R337H mutation carriers present different antioxidant enzyme activities and oxidative stress parameters when compared to non-carriers.
Effect of GDC-0449, a hedgehog (Hh) pathway inhibitor, on tumor activity and on erlotinib and cisplatin in a xenograft model of non-small cell lung cancer (NSCLC) with activated Hh pathway.
Authors: Ahmad Aamir , Sarkar Fazlul H. , Maitah Main , Ali Shadan , Sethi Seema , Aboukameel Amro , Gadgeel Shirish M.
Published: March 2020

e21057

Background: We have previously shown that chronic exposure of A549 NSCLC cell line to TGF-β1 can induce epithelial-mesenchymal transition (EMT) and can make these cell lines (A549-M) resistant to erlotinib, an EGFR inhibitor and cisplatin. EMT in A549-M cells was associated with significant activation of signaling through the Hh pathway. We therefore evaluated the activity of GDC-0449, an inhibitor of the Hh pathway, and assessed its activity in combination with erlotinib and cisplatin in A549-M cells both in vitro and in vivo. Methods: We assessed the effects of GDC-0449 (20 nM) on clonogenic growth (colony formation assay), cell motility (wound healing assay) and invasion (matrigel assay) in A549-M cells, and also assessed the activity of GDC-0449 in combination with erlotinib and cisplatin both in vitro (MTT assay) and in an experimental lung metastasis animal model in vivo by injecting A549-M cells through tail vein. Results: A549-M cells showed significantly higher rate of clonogenic growth, cell motility and invasion in vitro and a greater propensity to form metastases in vivo compared to parental A549 cells, and also showed higher resistance to erlotinib and cisplatin. A549-M cells showed higher mRNA levels of Shh (sonic hedgehog, a ligand of the Hh pathway) and ZEB1 and reduced levels of E-cadherin than A549 cells. GDC-0449 significantly reduced growth of A-549-M cell in vitro but had no effect on A-549 cells. GDC-0449 also reduced clonogenic growth, cell motility and invasion of A549-M cells. Pre-treatment with GDC-0449 significantly enhanced the activity of erlotinib (5 and 10 mM) and cisplatin (1mM-10mM) in A549-M cells (p < 0.05) but not in A-549 cells. GDC-0449 reduced the number of lung metastases in the xenograft model and enhanced the activity of erlotinib and cisplatin in this model. Conclusions: This data suggests that the EMT phenotype in NSCLC may be associated with the activation of the Hh signaling pathway. It also suggests that GDC-0449 has anti-tumor activity against NSCLC with activated Hh pathway, and can enhance the effects of erlotinib and cisplatin in such tumors.
Vysis ALK non-small cell lung cancer assay performance on archival tissue and cytology specimens.
Authors: Li Gu , Jacobson Kristine B , Matushek Paul E , Perez Thomas B , Herman Christine , Sachs Karen L , Pestova Ekaterina
Published: March 2020

e21060

Background: Rearrangements of ALK gene on chromosome 2p23 create a constitutively activated tyrosine kinase implicated in the development of NSCLC and targeted by novel therapeutic agents. The Vysis ALK Break Apart (BAP) FISH Probe assay was developed to detect chromosome 2p23 rearrangements in NSCLC FFPE tissue by fluorescence in situ hybridization. Performance of the ALK assay was assessed on FFPE tissue specimens and FFPE cell lines. The ALK probe was also tested on lung cytology cell suspensions. Methods: The ALK FISH assay success rate on primary NSCLC was tested with 30 FFPE tumor resection specimens, four 5-μm section slides each, judged by qualitative rating of fluorescent signal specificity, intensity, and background. Assay performance was further assessed on 15 FFPE biopsy specimens of 4 metastatic sites including mediastinal and axillary lymp nodes, pleural cavity, brain, and adrenal gland. Each specimen was tested in duplicate and evaluated by 2 reviewers. Assay reproducibility on FFPE cell pellets was tested with 3 lots of ALK Negative and ALK Positive Control Slides (cell lines) on 5 non-consecutive days. Slides were enumerated by 3 reviewers to determine % ALK positive cells, to a total of 45 results per control slide type. Additionally, the ALK BAP probe was tested with a modified assay protocol on ThinPrep lung cytology samples from 8 specimens. Results: The assay success rate on FFPE resection specimens was 99.2 % (90% CI 96.1% - 100%). The core biopsy specimens yielded 95% overall success rate. All Control Slides (FFPE cell lines) tested in the reproducibility study were enumerable within assay ranges, and were correctly classified per specifications of 0-8% and ≥20% ALK positive cells in ALK Negative and ALK Positive Control Slides, respectively. All of the lung cytology specimens prepared by ThinPrep hybridized successfully, however 2 specimens had low cellularity for ALK enumeration. Conclusions: The Vysis ALK BAP FISH assay demonstrated reproducible performance on FFPE cell lines and was shown to yield interpretable results on FFPE lung tissue specimens retrieved by resection or biopsy, from primary tumor or metastatic sites. Feasibility using lung cytology cell suspensions was demonstrated.
Ribonucleotide reductase subunit 2 (RRM2) to predict shorter survival in resected stage I-III non-small cell lung cancer (NSCLC) patients.
Authors: Rijavec Erika , Dal Bello Maria Giovanna , Savarino Graziana , Sini Claudio , Barletta Giulia , Genova Carlo , Truini Mauro , Pronzato Paolo , Merlo Domenico Franco , Pfeffer Ulrich , Grossi Francesco
Published: March 2020

e21065

Background: Biomarkers can help in identifying patients (pts) with early-stage NSCLC with high risk of relapse and poor prognosis. The aim of this study was to investigate the relationship between the levels of expression of 7 biomarkers, various clinicopathological characteristics and their prognostic significance. Methods: Tumor tissue from 82 radically resected stage I-III NSCLC pts were consecutively collected to investigate the mRNA expression and protein levels of the following biomarkers using quantitative reverse transcriptase real-time PCR (qRT-PCR) and immunohistochemistry (IHC) with a tissue microarray technique: excision repair cross-complementation group 1 (ERCC1), breast cancer 1 (BRCA1), ribonucleotide reductase subunit 1 (RRM1), RRM2, p53R2, thymidylate synthase (TS) and class III beta-tubulin (β-Tub-III). Results: On a univariate analysis, p53R2 expression was significantly higher in adenocarcinoma (ADK) compared to squamous cell carcinoma (SSC) samples (p=0.002) and in stage I compared to stage II-III (p≤0.001). ERCC1 expression was significantly higher in females compared to males (p=0.03), and β-Tub-III expression was significantly higher in ADK than in SSC (p=0.03). Pts with lower RRM2 expression survived longer than pts with higher RRM2 expression (p=0.069). The multivariate analysis confirmed RRM2 as an independent prognostic marker of shorter survival (p= 0.031). The comparison between survival curves with qRT-PCR and IHC showed similar results with a trend towards longer survival among ERCC1 negative pts, BRCA1 negative pts, p53R2 positive pts and among pts with low levels of RRM1 and RRM2, although the difference was not statistically significant with both methods. qRT-PCR and IHC have shown that β-Tub-III and TS had no significant impact on survival. Conclusions: This is the first study that identifies RRM2 expression as a negative prognostic factor in resected stage I-III NSCLC. Moreover, we have demonstrated the differential expression of p53R2 and β-Tub-III in ADK compared to SSC and higher expression of p53R2 in pts with stage I compared to stage II-III NSCLC.
Relationship of Ki67 to tumor size and lymph node metastasis in breast cancer.
Authors: Bonta Ioana , Bonta Dacian , Loch Michelle Marie , Eapen Ann , Blanchard Rita A.
Published: March 2020

e21076

Background: Ki67, a tumor proliferation marker, has demonstrated usefulness in breast cancer prognosis. Prior work with BrdU labeling for cell proliferation in breast cancer has not settled the question whether cell proliferation labeling is independent of other tumor features like tumor size and presence of axillary metastases, see Rew (1992) vs. Thor et al (1999). Methods: We analyzed retrospectively our database of 379 patients for correlation between the and tumor size, presence of axillary lymph node metastases and the percentage of Ki67 positve cells. We used linear and parabolic regression to correlate tumor size with the Ki67 index and receiver operator characteristics curve to correlate the presence of axillary lymph node metastases with the Ki67 index. Results: A very weak linear relationship was detected between tumor size and Ki67 index. The R-squared coefficient was 0.03, indicating that tumor size explained only 3% of the variability in measured Ki67 indexes. The ROC analysis, looking at the correlation between Ki67 and lymph node metastasis, yielded an area under the curve (Az) of 0.53. This indicates a very weak correlation. No relationship between axillary lymph node metastasis would yield an Az of 0.5 and a perfect correlation would yield an Az of 1. Conclusions: Tumor size and axillary lymph node metastasis explain less than 10% of observed Ki67 index variability. Therefore, in breast cancer, the prognostic value of Ki67 is additive to that of tumor size and lymph node involvement.
Prognostic significance of circulating tumor cell (CTC) levels at various times during first- and second-line chemotherapy for metastatic breast cancer (MBC).
Authors: Arteta-Bulos Rafael , Miller Paul J. E. , Overbury Rebecca S. , Schwartzberg Alison , Walker Mark S. , Schwartzberg Lee Steven
Published: March 2020

e21077

Background: CTCs have prognostic capability in the treatment of MBC based on a dichotomous level of ≥5/7.5 ml blood (Hi) vs. <5/7.5 ml blood (Lo) at baseline and first follow-up. The value of CTCs beyond baseline is not well established, and the implication of values that are reduced but remain high has not been explored. Methods: Chart review of patients (pts) on 1st or 2nd line treatment of MBC, with CTC determinations between 2/2008 and 9/2011, were retrospectively identified at a large community oncology practice. Pts with ≥2 CTCs or 1 CTC ≥5 were eligible. Demographics, treatment regimens and progression-free survival (PFS) were assessed. The Veridex Cell Search system was utilized for all CTC determinations. Qualifying baseline CTCs were those drawn -14 days to +30 days from start of treatment. Results: 221 eligible pts were identified, 71 with baseline CTCs in 1st line, and 84 in 2nd line. Median age was 59.5 (range 26-90), 40.7% were African-American, 17.6% HER2+, and 63%/49% ER/PR+. The median time to baseline CTC was 5 days in the 1st line and 1 day in the 2nd line. 1st line PFS was 8.0 vs. 5.8 mo among baseline Hi vs. Lo groups (p = 0.085). 2nd line PFS was 3.7 vs. 4.0 mo for Hi and Lo groups, respectively, (ns). Follow-up CTCs were obtained at clinical convenience. A 90 day landmark was used for analysis of PFS by four baseline/follow-up CTC pattern groups. 1st line PFS showed medians ranging from 6.0 mo (Hi/Hi) to 10.2 mo (Lo/Lo). In 1st line pts with Hi baseline CTCs (n=24), median PFS was 6.0 vs. 7.0 mo for Hi vs. Lo at follow-up (p = .64). Grouping pts with Hi baseline CTCs by reductions in absolute CTC count at follow-up (<33% vs. ≥33%) showed median PFS 3.5 mo for <33% reduction, 8.8 mo for ≥33% reduction (p = .0179). Sensitivity analyses with other cutoffs in reduction from 20-80% showed similar results. Conclusions: In the community setting, baseline and subsequent CTCs obtained at various times after baseline yield clinically useful prognostic information. Pts with positive baseline CTC levels experiencing a reduction of ≥33% exhibit significantly longer PFS, indicating that a simple high/low approach to the test can be modified to yield more prognostic value.
Pancreatic polypeptide as a biomarker for pancreatic neuroendocrine tumors.
Authors: Alistar Angela Tatiana , Moshier Erin L. , Kim Michelle , Warner Richard , Holcombe Randall F.
Published: March 2020

e21109

Background: Pancreatic neuroendocrine tumors (PNETs) represent tumors that secrete numerous proteins that could be used as biomarkers. We present a retrospective analysis of patients with PNETs who have had Pancreatic Polypeptide (PP) testing in an attempt to better define PP producing tumors as an entity and the role of PP as a biomarker. To our knowledge, this is the first single center comprehensive review of Pancreatic Polypeptide producing tumors. Methods: A retrospective study of patients with PNETs seen at Mount Sinai from 1980 to 2011. All patients that have had PP concentrations measured at least once were evaluated. Data relating to diagnosis, pathology, surgery, liver directed therapies, chemotherapy and survival outcome were noted. 71 patients with PNETs fulfilled the inclusion criteria. The 71 patients formed 4 subgroups. The 4 subgroups were evaluated and compared for survival: all 71 PP tested patients, 30 patients with PP producing tumors, 8 patients with PPomas, 22 patients with PP producing tumors that were non PP-omas and 41 patients with PP non-producing tumors Results: We identified a trend towards better survival for PNETs patients with PP producing tumors vs. non- PP producing tumors (p=0.19). There was no correlation between survival and a diagnosis of PPoma in relation to other PP producing tumors (p= 0.59) or non-PP producing tumors (p=0.58). Conclusions: Pancreatic Polypeptide is a biomarker that is worth prospective investigation and a standardized assay. Our analysis investigating Pancreatic Polypeptide as a prognostic and or predictive biomarker reveals a trend towards showing these characteristics. Using a standardized test and investigating this biomarker prospectively could lead to the validation of Pancreatic Polypeptide as a biomarker.
Molecular analysis of pancreaticobiliary fine needle aspirate/duct brushing cytocentrifugation supernatant fluid.
Authors: Finkelstein Sydney D. , Schmidt C. Max , Ellsworth Eric M. , Stearns Valerie T. , Smith Dennis M.
Published: March 2020

e21121

Background: Reliance on minimally invasive sampling techniques (fine needle aspiration (FNA), duct brushing) continues to increase for pancreaticobiliary (PB) pre-cancer/cancer diagnosis. Fundamental limitations include low cellularity, sampling variability, and indeterminate microscopic diagnosis. To address these issues, we developed a novel approach involving DNA mutational profiling of the soluble (or free) DNA in the cytocentrifugation supernatant fluid (supernatant) currently discarded during cytology preparation. Methods: 77 PB specimens underwent combined microscopic/molecular analysis using free DNA in cytology supernatant fluid. Pancreatic FNA and biliary cytobrushes had either surgical pathology or positive cytology as the gold standard. DNA was extracted from 2 mL of the supernatant and from corresponding microdissected cytology cells. Profiling included DNA quantity and quality, and analyses for KRAS mutations and 16 LOH (loss of heterozygosity) mutations. Results: The cell-free supernatant showed significantly higher levels of more intact amplifiable DNA than typical microdissection specimens. All non-neoplastic specimens (n=38) showed no false positive mutations. 37/39 malignant specimens showed mutations (sensitivity 95%, specificity 100%). Mutational profiles of the microdissected cells and the supernatant demonstrated high concordance. However, mutation detection was consistently superior in the supernatant fluid (mean 6.7 mutations) compared to the microdissected cells (mean 4.2). Mutation clonality was also significantly higher in the supernatant. Conclusions: Despite the lack of cells in many PB samples, the post-centrifugation supernatant fluid provides an abundant, robust, and highly representative source of DNA for molecular analysis. Of note, the greater extent and clonality of detected mutations in the supernatant fluid supports it being enriched with neoplastic DNA when cancer is present. This novel approach complements standard cytology practice by increasing sensitivity and specificity of cancer detection while reducing sampling variation.
Adequacy of biopsy samples for molecular testing in patients with non-small cell lung cancer.
Authors: Hrinczenko Borys , Subramonia-Iyer Subramony , Zhang Deng , Olsen Brian
Published: March 2020

e21133

Background: Testing for a mutated EGFR gene has transformed the care of NSCLC. Although ASCO recommends EGFR mutation testing for cases of NSCLC, there have been no studies to assess adequacy of biopsy samples for molecular testing in non-trial settings. Methods: A retrospective chart review to identify patients diagnosed with NSCLC at our community hospital during 2010. A pathologist then reviewed the adequacy of archivedbiopsy material for EGFR mutation testing based on ASCO criteria (Eberhard, JCO 2008). Results: In 2010, 64 cases of NSCLC were diagnosed. Demographics:Thirty-six (56%) of the patients were male; median age was 69.5 years, 27 were current smokers, 33 were ex-smokers, median survival was 4.9 months. Histologic subtypes: squamous - 28 (44%), adenocarcinoma - 23 (36%), bronchoalveolar - 3 (5%), adeno with mixed bronchoalveolar - 2 (3%), large cell - 3 (5%), and large cell neuroendocrine - 2 (3%). Twenty-eight (44%) were considered adenocarcinoma. Staging: Stage I, II, III, IV - 18 (28%), 2 (3%), 13 (20%), 23 (36%). Eight (13%) had incomplete staging information. Biopsy material: transbronchial FNA in 23 (36%), CT-guided FNA in 13 (20%), definitive surgery in 18 (28%), other - 10 patients (15%). Imaging review: 29 central lesions (45%) and 35 (55%) peripheral. Specimen review: 45/64 (70%) were adequate for EGFR mutation analysis; Twenty cases (30%) were not. Among adenocarcinoma, 19/28 (68%) cases had adequate samples, but 9 (32%) did not. Conclusions: In this series, nearly one-third of all NSCLC cases, including adenocarcinoma, did not yield sufficient specimen from the first biopsy for EGFR mutation analysis. Newer molecular markers (EGFR, EML4-ALK) with targeted therapy such as erlotinib and crizotinib are available in NSCLC so adequate tissue sampling is imperative. Lack of satisfactory tumor tissue sampling may preclude mutation testing thereby denying these patients treatment with potential benefit and lower toxicity. This has implications for planned diagnostic investigations since an appropriate site must be biopsied so as to obtain an ample sample. Efforts to improve clinician awareness about adequate tumor sampling are needed and should be a top priority for multidisciplinary lung cancer teams.
Breast cancer in a patient with multiple endocrine neoplasia type 1 (MEN 1): A case report and review of the literature.
Authors: Inic Zorka Momcilo , Inic Momcilo , Dzodic Radan , Pupic Gordana , Damjanovic Svetozar
Published: March 2020

e21136

Background: We treated a patient with breast cancer associated with multiple endocrine neoplasia type 1 (MEN 1). There are only few studies of the development of breast cancer in patients diagnosed with MEN1 syndrome. Methods: We found that patient had a pituitary macroadenoma that secretes prolactin (prolactinomas), increased serum concentration of calcium, 4.09 mmol/l and increased serum concentration of PTH, 212 pg/ml. Neck ultrasound and scintigrafy showed suspicious adenoma in lower left parathyroid gland. A computed tomography scan of the abdomen revealed a tumor mass in the body of the pancreas and adenoma in the left suprarenal gland. We diagnosed multiple endocrine neoplasia type 1 (MEN 1). Results: MEN1 occurs as a result of inactivating mutations of the MEN1 gene (MEN1), located on chromosome11q13. Results indicate that menin is a direct activator of ERalpha function. In a clinical study, in Kagawa, with 65 ER-positive breast cancer samples-all of which had been treated with tamoxifen for 2-5 years as adjuvant therapies-menin-positive tumors had a worse outcome than menin-negative ones. This indicated that menin can function as a transcriptional regulator of ERalpha and is a possible predictive factor for tamoxifen resistance.These results demonstrate that allelic deletions of the 13q12-14 region occur in some pituitary adenomas and 16% of parathyroid adenomas. Conclusions: As recent molecular studies have suggested genetic mutations in multiple endocrine neoplasia type 1, this syndrome may possibly predispose patients to breast cancer.This possibility cannot be evaluated definitively on the basis of a single case report; additional observations and studies are necessary to investigate this hypothesis further.
Pharmacodynamic (PD) evaluation of soluble proteins from a phase I trial of lapatinib (L) in combination with sorafenib (S).
Authors: Simonelli Matteo , Zucali Paolo Andrea , Lorenzi Elena , Rubino Luca , De Vincenzo Fabio , Perrino Matteo , Rimassa Lorenza , Giordano Laura , Santoro Armando
Published: March 2020

e21139

Background: At ASCO 2011 we presented safety and efficacy results from a phase I study of L in combination with S in patients with solid tumors refractory to standard therapy (NCT00984425). To investigate potential biomarkers of biological activity of this combination, we analyzed serum levels of a panel of soluble proteins characterizing the oncogenic signalling pathways targeted by L and S (VEGF, sVEGFR-2, EGF, sEGFR, sHER2/neu), and others mediators of angiogenesis such as sPDGFR, IL-8, IL-6, bFGF, PIGF, TGF-α e β, HGF. Methods: Serum samples from 22/30 patients enrolled in this trial were prospectively collected at baseline and then every two months until treatment discontinuation. Serum levels of VEGF, sVEGFR-2, EGF, sEGFR, sHER2/neu and sPDGFR were measured via enzyme-linked immunoadsorbent assays analysis (ELISA). Possible correlations between serum proteins levels and disease-control rate (SCR), PFS or OS were assessed by logistic and Cox regression model, respectively. Results: Analysis of baseline biomarkers as a function of demographic variables indicated that VEGF levels were inversely correlated with age (P= .018) and VEGFR-2 was significantly associated with tumor burden (P= .037). Risk of progression increased with high baseline sEGFR levels (HR 1.10; p= .0498). Overall, PD changes suggest that treatment with L and S increased VEGF and sHER2/neu levels, while decreased EGF, sEGFR, VEGFR-2 and sPDGFR. Mainly, sEGFR levels decreased by at least 75% at the first radiological assessment in half of patients, whereas at the end of treatment a reduction ≥80% was observed in 18 patients; a greater reduction of sEGFR levels was significantly associated with a higher risk of progression (HR 8.3; P <.01) and death (HR 3.6; P= .023). An increase of VEGF levels during treatment seemed to be associated with better SCR (OR: 6.33; P= .085). Conclusions: In this small subset of patients treated with L and S a significant modulation of serum biomarkers was observed. The clinical significance of these changes as potential surrogate markers requires further evaluation. Data concerning other angiogenic markers along with tissue analysis (IHC, FISH and mutation) will be presented at the meeting.
Positron emission tomography/computed tomography (PET/CT) as a biomarker of pathologic response to neoadjuvant chemotherapy in breast carcinoma.
Authors: Kelly Catherine M. , Smith Clare , Conlon Susan , Salman Reem , McCaffrey John , Jordan Emmet
Published: March 2020

e21147

Background: Pathological complete response (pCR) after neoadjuvant chemotherapy (NAC) in breast carcinoma is prognostic. Predictive biomarkers for pCR include early response to NAC, estrogen receptor (ER) negativity, HER2 positivity, and high Ki67. We assessed whether absence of fluoro-deoxy glucose (FDG) uptake measured by standardized uptake value (SUV) after NAC would predict pCR. Methods: We identified 23 patients (pts) who had PET/CT scanning pre and post NAC. We examined breast cancer subtype, chemotherapy (CT) regimen, number of cycles of CT given, clinical and pathological staging data and changes in SUV in the breasts and lymph nodes pre and post NAC. pCR was defined as no residual cancer in the breast or axillary lymph nodes. Results: Median age at diagnosis was 46 years (IQR; 37 to 56). Median tumor size at diagnosis was 30mm (IQR; 25 to 43) and 19 pts (83%) had node positive breast cancer. Most tumors were ductal (n=22) with 1 lobular cancer. Preoperatively 95% received all CT. All HER2+ pts received Trastuzumab. Anthracycline/taxane based regimens were most frequently given in 22 cases, 1 received lapatinib/trastuzumab. Five tumors (21.7%) were ER+/HER2+; 14 (60.9%) ER+/HER2-; 2 (8.7%) ER-/HER2+ and 2 (8.7%) were ER-/HER2-. All tumors were high (n=9, 39.1%) or intermediate grade (n=14, 61%). SUV was significantly lower post NAC (p=0.035). We observed no SUV uptake in breast or lymph nodes in 15 cases (65.2%) post NAC, these corresponded to; ER+HER2+ 4/5 (80%); ER+HER2- 7/15 (46.7%); ER-HER2- 2/2(100%), ER-HER2+ 2/2(100%). Absent SUV uptake post NAC was associated with a pCR (breast and lymph nodes) in 5/15 (33%) of pts (ER+HER2+ n=1, ER+HER2- n=1, ER-HER2- n=2, ER-HER2+ n=1). Ten of 15 tumors (67%) had no SUV uptake in the breast post NAC and 7 (47%) were associated with a pCR. There was a trend toward increased odds of pCR with no SUV uptake post NAC (OR 2.76; 95% CI 0.85 to 8.94: P= 0.09). Overall rate of pCR was 21.7% (n=5). Conclusions: A non-statistically significant trend toward increased odds of pCR with no SUV uptake post NAC was observed. Larger subtype-specific breast cancer cohorts will be required to determine the value of PET/CT as a predictive biomarker for pCR.
Expression of CK-19, galectin-3, and HBME-1 in the differentiation of thyroid lesions: Systematic review and DIAGNOSTIC meta-analysis.
Authors: Matos Leandro Luongo , Del Giglio Adriana Braz , Ogawa Carolina MATSUBAYASHI , Farah Michelle de lima , Pinhal Maria Aparecida Silva , Del Giglio Auro
Published: March 2020

e21151

Background: The distinction between malignant and benign lesions of the thyroid gland often requires histological demonstration, once the fine-needle aspiration biopsy method applied pre-operatively has some limitations. In an attempt to improve diagnostic accuracy, markers have been studied by immunocyto- and immunohistochemistry techniques, mainly cytokeratin-19 (CK-19), galectin-3 (Gal-3) and Hector Battifora mesothelial-1 (HBME-1). However, current results remain controversial. The aim of the present article was to establish the diagnostic accuracy of CK-19, Gal-3 and HBME-1 markers, as well as their associations, in the differentiation of malignant and benign thyroid lesions. Methods: A systematic review of MEDLINE’s published articles was performed. After establishing the inclusion and exclusion criteria, 65 articles were selected. The technique of meta-analysis of diagnostic accuracy was employed, and global values of sensitivity, specificity, area under the ROC curve (AUC), and diagnostic odds ratio (dOR) were calculated. Results: The positivity ofCK-19 for the diagnosis of malignant thyroid lesions demonstrated sensitivity of 81% (IC95%: 79-83%), specificity of 73% (IC95%: 70-75%), AUC of 87,5% and dOR of 14,7 (IC95%: 8,2-26,4); for Gal-3, sensitivity of 82% (IC95%: 81-84%), specificity of 81% (IC95%: 79-82%), AUC of 90% and dOR of 23,4 (IC95%: 14,0-39,1); and for HBME-1, sensitivity of 77% (IC95%: 76-79%), specificity of 83% (IC95%: 82-85%), AUC of 92,8% and dOR of 41,0 (IC95%: 21,4-78,4). The association of the three markers determined sensitivity of 85% (IC95%: 78-90%), specificity of 97% (IC95%: 90-99%), AUC of 93,3% and dOR of 95,1 (IC95%: 25,2-359,1). Conclusions: The search for new molecular markers must continue in order to enhance the diagnostic accuracy of the differentiation between malignant and benign lesions of the thyroid gland, since the results found still show persistency of false-negative and false-positive tests.
NSABP B-38: Definitive analysis of a randomized adjuvant trial comparing dose-dense (DD) AC followed by paclitaxel (P) plus gemcitabine (G) with DD AC followed by P and with docetaxel, doxorubicin, and cyclophosphamide (TAC) in women with operable, node-positive breast cancer.
Authors: Swain Sandra M. , Tang Gong , Geyer Charles E. , Rastogi Priya , Atkins James Norman , Donnellan Paul P. , Fehrenbacher Louis , Azar Catherine A. , Robidoux Andre , Polikoff Jonathan , Brufsky Adam , Biggs David D. , Levine Edward A. , Zapas John L. , Provencher Louise , Perez Edith A. , Paik Soonmyung , Costantino Joseph P. , Mamounas Eleftherios P. , Wolmark Norman
Published: March 2020

LBA1000

The full, final text of this abstract will be available at abstract.asco.org at 12:01 AM (EDT) on Monday, June 4, 2012, and in the Annual Meeting Proceedings online supplement to the June 20, 2012, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Monday edition of ASCO Daily News.