bluuebunny/crossref_metadata_2025 huggingface dataset:
Factors associated with survival in non-small cell lung cancer (NSCLC) patients with a solitary metastasis.
Authors: Parikh Ravi , Cronin Angel , Kozono David E. , Oxnard Geoffrey R. , Mak Raymond H. , Jackman David Michael , Lo Peter C. , Baldini Elizabeth H. , Johnson Bruce E. , Chen Aileen B.
Published: March 2020

e19121

Background: Although palliative chemotherapy is the standard of care for metastatic NSCLC, somepatients with oligometastatic disease may benefit from aggressive local therapy. We investigated factors associated with greater survival among patients diagnosed with a solitary metastatic lesion. Methods: We identified patients diagnosed with stage IV NSCLC who presented with a solitary metastatic lesion based on PET and MRI and who were prospectively consented and enrolled in our institutional database from 2002-2011. Univariable and multivariable Cox proportional hazards models were used to analyze factors associated with overall survival among this cohort. Results: We identified 110 patients (10.7% of stage IV patients) meeting our inclusion criteria. Median age at diagnosis was 61 years, 50% of patients were female, 66% had adenocarcinoma histology, and 35% had N0-1 disease. Median survival from diagnosis was 18.7 months, with a median followup of 31.5 months. On univariable analysis, greater overall survival was associated with ECOG performance status 0-1 vs 2+ (median 21.5 months vs 12.6 months, HR 0.32, p<0.01); weight loss <2 vs >2 kg (22.4 vs 13.8, HR 0.56, p=0.03); and N stage 0-1 vs 2-3 (32.0 vs 17.6, HR 0.52, p=0.02). Adenocarcinoma vs non-adenocarcinoma histology (22.9 vs 13.8, HR 0.65, p=0.07) was borderline significant. Age, gender, race, current smoking, size of primary tumor, and metastatic organ were not significantly associated with survival. On multivariable analysis, adenocarcinoma histology (HR= 0.58, p=0.06); N stage 0-1 (HR= 0.43, p=0.01); and weight loss <2 kg (HR 0.53, p=0.03) were associated with greater overall survival. Conclusions: Select patient and tumor characteristics may predict for improved survival among patients with oligometastatic NSCLC. Future studies will evaluate the impact of aggressive local therapy in these patients.
A survey on Japanese thoracic oncologists’ preference on treatment strategy for EGFR-mutant or EML4-ALK-mutant non-small cell lung cancer (NSCLC).
Authors: Hotta Katsuyuki , Kiura Katsuyuki , Tabata Masarhiro , Takigawa Nagio , Tanimoto Mitsune , Ueoka Hiroshi
Published: March 2020

e19124

Background: NSCLC pts have become categorized based on two genetic alterations in their tumors, EGFR and EML4-ALK, to provide personalized therapy. In EGFR-mutant NSCLC, EGFR-TKI yields a comparable survival with platinum-based chemotherapy, suggesting either of them can be initiated first, and its choice seems to depend partly on oncologists’ preference. As for the other, EML4-ALK, NCCN guideline recommends first-line crizotinib use despite no relevant survival data. The decision whether to choose crizotinib might be also influenced by oncologists’ preference. We here assessed their preference. Methods: The Japanese thoracic oncologists were asked to complete a self-administered questionnaire at the Japanese meeting specific for thoracic oncology. Results: Of 3,046 subjects, 871 (29%) responded voluntarily to a survey, mainly consisting of medical and surgical oncologists (93%). Majority considered EGFR mutation status should be checked at the time of diagnosis (89%), whereas only 60% and 33% reported EML4-ALK gene status would be assessed at the time of diagnosis and until the initiation of second-line setting, respectively. The subjects also considered pts with EGFR-wild-typed tumor and pts with clinical characteristics possibly related to EML4-ALK rearrangement should selectively receive EML4-ALK gene status check rather than all NSCLC pts (57%, 22% and 16%, respectively). Among the subjects, 52% preferred to choose gefitinib rather than platinum in the first-line setting in EGFR-mutant NSCLC, whilst 44% preferred crizotinib to platinum in the first-line setting in EML4-ALK-positive NSCLC. The major reasons why they chose gefitinib in EGFR-mutant NSCLC were ‘PFS is better’ (36%) and ‘it is easy to improve QOL’ (25%), whereas ‘PFS is better’ (43%), and ‘I want to prescribe when patients are still in good condition’ (19%) were the predominant reasons for choosing crizotinib in EML4-ALK-positive NSCLC. Conclusions: About half of the subjects preferred each molecular-targeted agent to the conventional cytotoxic chemotherapy in first-line setting. They considered better PFS was important in the treatment of EGFR-mutant or EML4-ALK-mutant NSCLC.
Immunohistochemistry to detect EGFR mutation and ALK gene rearrangement in NSCLC bronchoscopic specimens.
Authors: Wang Mengzhao , Pan Qing qing , Zhu Pei
Published: March 2020

e19135

Background: The discovery of the E746-A750 and L858R antibodies for mutated epidermal growth factor receptor (EGFR) and the D5F3 antibody for the product of the rearranged anaplastic lymphoma kinase (ALK) gene enables immunohistochemistry(IHC) to be applied to the identification of lung cancer. This study aimed to explore the effectiveness of IHC in small specimens and compare it with standard methods. Methods: We collected bronchoscopic specimens of lung adenocarcinoma that were confirmed by pathology; gathered clinical information, including the basic characteristics of patients, staging and differentiation of the tumor, and the amount and proportion of tumor cells; identified mutant EGFR with E746-A750del and L858R antibodies and rearranged ALK with D5F3 antibody (Cell Signaling Technology, USA); and compared their sensitivity and specificity with standard methods. Results: We collected 92 patients¡&hibar; bronchoscopic specimens. All specimens were successfully detected by the Scorpions-ARMS method£€and exon 19 deletion occurred in 15 specimens (16.3%), and exon 21 L858R mutation in 15 specimens (16.3%). In EGFR-IHC, the positive rates of E746-A750del and L858R were 17.8% (16/92) and 21.4% (19/92), respectively. The sensitivity and specificity of E746-A750del (with Scorpions-amplification refractory mutation system (ARMS) as the standard) were 73.3% (11/15) and 93.3% (70/75), respectively, and those of L858R were 93.3% (14/15) and 93.2% (69/74). Neither method was significantly worse than Scorpions-ARMS (p£½1.000 for E746-A750del, p£½0.219 for L858R). FISH detected 6 positive and 71 negative specimens for ALK rearrangement; the other 15 could not be evaluated for various different reasons. In ALK-IHC, the positive rate was 7.8% (6/77; 13 could not be evaluated). The sensitivity and specificity of D5F3 were 50% (3/6) and 100% (71/71), respectively. ALK-IHC showed no difference with the standard method of fluorescence in situ hybridization (FISH). Conclusions: IHC with E746-A750del and L858R antibodies is an economical and convenient method for EGFR mutation detection in small specimens of NSCLC, as is D5F3 monoclonal antibody for ALK-IHC, especially for the primary screening.
Pulmonary blastoma: A clinicopathologic study of three cases.
Authors: Sandoval-Sus Jose D , Hansra Damien Mikael , Cioffi Lavina Maureen , Patel Ravi , Ponce Mario , Santos Edgardo S , Ikpatt Offiong F
Published: March 2020

e19140

Background: Pulmonary blastoma (PB) is a rare biphasic neoplasm representing 0.2-0.5% of pulmonary malignancies. Presentation ranges from asymptomatic to symptoms of advanced pulmonary tumors. Surgery is the preferred treatment with no consensus on the role of other adjuvant therapies. Methods: We report the clinicopathologic features of PB in 3 patients (pts). Retrospective review of medical records of pts diagnosed with PB at Jackson Memorial Hospital, Miami, FL since 2009. Staging performed using the 2009 AJCC system for NSCLC. Cases required pathological diagnosis before inclusion. Variables analyzed were demographics, clinicopathologic presentation, treatments and outcomes. Results: Mean age at diagnosis was 53 years old; 2 were male; Hispanic and Caucasian and 1 African American female. Both males were heavy smokers. At diagnosis, all cases were symptomatic; 1 had hypercalcemia of 12.9 mg/dL and 2 were stage IV with metastatic disease (bone and thyroid). 2/3 where treated with surgical resection and both achieved complete remission (CR); Pt with stage IV and resection relapsed in 28 months, received chemotherapy (3 cycles cisplatin/docetaxel) with progressive disease then radiation (37 Gy) without response and died. The other surgical patient remains in CR. The patient with unresectable disease received chemotherapy (6 cycles carboplatin/docetaxel) attaining partial remission. 2/3 patients remain alive at end of this study. Pathologically, all cases showed biphasic morphology composed of multiple glandular structures with atypical pseudostratified columnar epithelium and cellular stromal component. Immunohistochemical analysis revealed positivity in all cases for thyroid transcription factor (TTF-1), keratin, and epithelial membrane antigen (EMA) in the glandular component. Conclusions: All pts were symptomatic at diagnosis and 2 presented with metastatic disease. All pts had similar morphologically and immunophenotypically. Surgery was the most beneficial intervention in regard to CR and PFS. Chemotherapy has shown mixed responses, thus, larger studies are needed to define the role of chemotherapy in the adjuvant and palliative settings.
A phase I feasability study of concurrent fondaparinux (F) with carboplatin (Crb) and paclitaxel (P) for metastatic non-small cell lung cancer (NSCLC): An analysis of coagulation/angiogenic biomarker (CABM) kinetics.
Authors: Mooney David James , Miley Debi , Jerome Mary , Grant Stefan C. , Robert Francisco
Published: March 2020

e19143

Background: There is an association between risk of thrombosis and metastatic cancer. Chemotherapy (C) also independently increases thrombotic risk. This increased risk is multifactorial, including endothelial damage and release of angiogenic cytokines. We hypothesized that adding anticoagulation to C may decrease thrombotic risk and also, potentially, have anti-tumor effect. Methods: The primary aim of this study was to determine the tolerability and safety (bleeding events) of the combination of F with 21-day cycle chemotherapy (Crb AUC 6 + P 200 mg/m2) in two cohorts of untreated patients (pts) with stage IV NSCLC. The secondary objectives were to determine incidence of venous thrombosis (VT), changes in CABM parameters during treatment, and clinical efficacy endpoints. Two cohorts of pts received F from cycles 2-4 with Crb+P. Cohort A received 2.5 mg F daily from cycle 2-4. Cohort B received 7.5 mg of F on day 1, 2 of cycle 2-4 and 2.5 mg F on day 3-21. Results: Clinical data from 19 evaluable pts are as follows: median age 55 years, 63% male, and 32% adenocarcinoma. There was no major bleeding event (BE) in either cohort, and 5 pts had a minor BE. There was no VT. Median time to progression was 5 months (3.8-6.2 months), and overall survival was 10 months (4.3-15.6 months). Baseline values of sensitive markers of activated coagulation (D-Dimer, Thrombin Anti-Thrombin Complex) were above the normal range in most patients. These biomarkers tended to increase during the first cycle (without F); whereas the same markers decreased during the second cycle (with F). A reduction of the angiogenic biomarkers during therapy was observed with VEGF, TGF-β1, and Angiopoietin-1. Conclusions: Concurrent treatment with F and chemotherapy for metastatic NSCLC is feasible with no major bleeding and little minor bleeding. During chemotherapy, coagulant and angiogenic biomarkers tended to increase, perhaps suggesting an increase in thrombogenic state. When F was added, these markers trended downward, suggesting that the proangiogenic state associated with cancer may be significantly altered by anticoagulation. Clinical trial information: NCT00476216.
Risk of interstitial lung disease with gefitinib and erlotinib in advanced non-small cell lung cancer: A systematic review and meta-analysis of clinical trials.
Authors: Shi Liang , Liu Zhe , Tang Junfang , Tong Li
Published: March 2020

e19149

Background: Gefitinib and erlotinib are oral EGFR-TKIs widely used in advanced NSCLC. The reported incidence and risk of interstitial lung disease (ILD) events associated with gefitinib and erlotinib varies widely and has been inconsistently reported in trials. Therefore, we performed a systematic review and meta-analysis to determine the incidence and the relative risk (RR) associated with the use of gefitinib and erlotinib. Methods: PubMed databases were searched for articles published from January 2000 to October 2012, and abstracts from ASCO and ESMO meetings (2000-2012) were searched for relevant studies. Eligible studies included randomized controlled trials with gefitinib and erlotinib in advanced NSCLC patients. Summary incidence rates, RRs, and 95% CIs were calculated using fixed-effects or random-effects models, depending on the heterogeneity of the included studies. Results: 15,618 patients from 29 randomized controlled trials were selected for this meta-analysis. The overall incidence for all-grade ILD events was 1.2% (95% CI, 0.9% to 1.6%) among patients receiving gefitinib and erlotinib, with a mortality of 22.8% (95% CI, 14.6% to 31.0%). The summary incidence of fatal ILD events was 0.2% (95% CI, 0.1% to 0.3%) in 26 trials with available data. Compared with controls, the RR of all-grade and fatal ILD events was 1.53 (95% CI, 1.13 to 2.08; P = 0.006) and 1.96 (95% CI, 1.03 to 3.72, P = 0.041), respectively. After stratifying patients by study location (Asian versus non-Asian), the RR of all-grade ILD events was 2.00 (95% CI, 1.23 to 3.25) and 1.30 (95% CI, 0.88 to 1.91) for Asian studies and non-Asian studies, respectively. The analysis was also stratified for drug type, trial phase, treatment line, and studies quality, but no significant differences in RRs were observed in all subgroup analyses. Conclusions: Treatment with EGFR-TKIs gefitinib and erlotinib is associated with a significant increase in the risk of developing both all-grade and fatal ILD events in advanced NSCLC.
Efficient detection of EGFR, KRAS, and BRAF mutations in cell-free DNA from pleural effusions.
Authors: Sagan Christine , Vallee Audrey , Denis Marc G.
Published: March 2020

e19151

Background: Detection of EGFR gene alterations is critical for predicting the response to tyrosine kinase inhibitors in patients with non-small-cell lung cancer. Analysis of other biomarkers might also be mandatory in the future, requiring more biological material. In routine practice, molecular testing is usually performed on tissues such as bronchial or transthoracic biopsies and surgical resections. We investigated the feasibility of using both cytology cell blocks and cell-free DNA from pleural effusions. Methods: Pleural effusion samples were centrifuged to form a pellet, fixed in formaldehyde, and processed as a cell block. DNA was extracted from 10-µm sections after paraffin removal using the Forensic kit and an iPrep system (Invitrogen). Cell-free DNA was extracted from the centrifugation supernatants using the QIAamp Circulating Nucleic Acid kit (Qiagen). Detection of molecular alterations were performed by allele-specific PCR (EGFR p.L858R mutation, BRAF p.V600E mutation), fragment size analysis (EGFR exon 19 deletions), and Sanger sequencing (KRAS mutations, codons 12 and 13). Results: Between January 2010 and December 2012, we tested 65 cell pellets (32 women and 33 men) and collected 34 paired supernatants. With the cell pellets, all the EGFR status were successfully determined (13 mutations; 20%), 9 mutations of KRAS on the 42 cases sequenced (17.6%), and 2 BRAF mutations on the 50 samples tested (4%) were found. We next investigated the potential use of the cell-free DNA from supernatants. Quantification by real-time PCR revealed significant amounts of DNA (mean = 306 ng extracted from 2 ml). EGFR, KRAS and BRAF tests were contributive for all these samples: 2 EGFR p.L858R mutations, 6 KRAS mutation (p.G12C, n=2; p.G12D, n=2; p.G13C, n=1 and p.G12F, n=1) and 1 BRAF p.V600E mutation were found. Most interestingly, the molecular status of the cell-free DNAs perfectly matched those obtained with the corresponding cell pellets. Conclusions: Pleural effusions provide high quality material for molecular testing and are suitable samples for routine practice. Cell-free DNA can also be successfully tested thus saving material for morphological / immunohistochemical / FISH analysis.
Clinical significance of ERCC2 haplotype-tagging single nucleotide polymorphisms in patients with unresectable non-small cell lung cancer treated with first-line platinum-based chemotherapy.
Authors: Kim Seok-Hyun , Lee Gyeong-won , Jeong Yi Yeong , Kim Ho-Cheol , Lee Jong Deog , Hwang Young Sil , Kim In-Suk , Lee Suee , Oh Sung Yong
Published: March 2020

e19156

Background: DNA repair pathway is a crucial molecular mechanism potentially involved in resistance to platinum-based chemotherapy in the unresectable non-small cell lung cancer. Furthermore, single nucleotide polymorphisms (SNPs) and haplotypes in the ERCC2 gene are thought to be associated with the risk of developing lung cancer and clinical outcomes. Methods: We genotyped 8 ERCC2 htSNPs for 129 unresectable NSCLC (stage IIIA, 12; stage III, 36; stage IV, 82) cases treated with first-line platinum-based chemotherapy. Clinical characteristics, treatment outcomes, and predictive value of htSNPs in patient response, survival, and chemotherapy-related adverse events were analyzed according to each ERCC2 htSNP using chi-square test, Kaplan-Meier method, and Cox proportional hazard model. Results: No differences were observed in patient or disease characteristics and response according to ERCC2 htSNPs. In a survival analysis, rs50872 was significantly related to overall survival (OS) (log-rank test, p=0.014). The median survival duration of rs50872 G/G, A/G, and A/A genotypes was 35.75 (95% confidence interval [CI] 21.05-50.45), 36.07 (hazard ratio[HR] 1.02, 95% CI 25.20-46.94), and 16.75 (HR 3.49, 95% CI 5.73-27.77) months, respectively. A significant association was observed between grade 3-4 infections and poor survival: OS in patients with a grade 0-2 infection: 35.75 months (95% CI 28.15-43.35); OS in patients with a grade 3-4 infection: 12.86 months (95% CI 8.99-16.72, HR 3.57) (log-rank test, p<0.001). In a subgroup analysis, rs238405 genotype was significantly related to OS in the taxane-based group. However, rs238416 genotype was significantly associated with OS in the gemcitabine-based group. Conclusions: ERCC2 htSNPs rs50872 (overall), rs238405 (taxane-based group), and rs238416 (gemcitabine-based group) and infection related to first-line chemotherapy were associated with OS in unresectable NSCLC patients treated with first-line platinum-based chemotherapy. However, additional large prospective studies focusing on the role of ERCC2 htSNPs in unresectable NSCLC are needed.
Evaluation of spot urine protein to creatinine ratio and 24-hour urine protein quantification for proteinuria in patients with plasma cell dyscrasia.
Authors: Apuri Susmitha , Nishihori Taiga , Lin Karen Yin , Baz Rachid C. , Kim Jongphil , Shain Kenneth H. , Doumit Elias , Ochoa-Bayona Jose L. , Sullivan Daniel , Alsina Melissa , Djulbegovic Benjamin
Published: March 2020

e19516

Background: Though cumbersome, a 24-hour urine collection remains crucial for diagnosis and monitoring in plasma cell dyscrasias (PCD). Nephrology guidelines recommend replacement of 24-hour urine collection with spot urine protein/creatinine (Pr/Cr) ratio for proteinuria. Only limited data exist on accuracy of spot urine Pr/Cr ratio in PCD. Methods: From 04/2012 to 01/2013, a total of 67 PCD patients were prospectively enrolled. Oliguria or dialysis were considered ineligible. Spot urine was collected on the day of 24-hour urine collection. Spot urine Pr/Cr ratios were compared to 24-hour urine on (1) total protein and (2) monoclonal protein (M-spike). Results: Fifty-two of 67 patients were evaluable (missing samples in 9; no Pr/Cr ratios in 4; protein below the level of detection in 2). A median age was 66 (range, 36 - 82), 62% were male, and 81% were Caucasian. Primary diagnoses were myeloma (n=47), amyloidosis (n=2), plasmacytoma (n=2), and MGUS (n=1). Comorbidities included hypertension (58%), chronic kidney disease (27%), diabetes (19%), and congestive heart failure (2%). A median serum creatinine was 0.9 mg/dL (range, 0.5 - 5.1). A median spot urine Pr/Cr ratio was 140 mg/g creatinine (range, 32 - 21,447), a median 24-hour urine protein was 130 mg (range, 31 - 15,092: n=49), and a median urine M-spike was 1.9 mg (range, 0 - 8,099). Most spot urine samples were collected in the morning (65%). There were strong correlations between (1) spot urine Pr/Cr ratio and 24-hour total urine protein (Spearman correlation coefficient=0.91, p < 0.0001), and (2) spot urine Pr/Cr ratio and 24-hour urine M-spike (Spearman correlation coefficient=0.73, p < 0.0001). The timing of spot urine sample collection had no significant effect (p = 0.274 by Wilcoxon rank-sum test). Conclusions: Spot urine Pr/Cr ratio strongly correlates with proteinuria measured in 24-hour urine collection and may predict the quantity of urine M-spike in non-oliguric PCD population. Spot urine Pr/Cr ratio may be useful as a monitoring tool for estimation of proteinuria. The study is ongoing and updated result will be provided.
Lack of effect of bendamustine on Fridericia corrected QT interval (QTcF) in a phase III study of bendamustine-rituximab in advanced indolent non-Hodgkin’s lymphoma (NHL) or mantle cell lymphoma (MCL).
Authors: Burke John M. , Flinn Ian , Craig Michael , Van Der Jagt Richard H. C. , MacDonald David , Hallman Doreen M. , Munteanu Mihaela C.
Published: March 2020

e19518

Background: Clinical data on the arrhythmogenic potential of bendamustine are limited. Bendamustine and rituximab (BR) are commonly combined, and rituximab concentrations are essentially constant ≥1.5 hours due to its long half-life. This BR single-arm study assessed the effects of bendamustine on QTcF over 1.5 hours in previously untreated adults with advanced indolent NHL or MCL. Methods: Patients in this open-label study were aged ≥18 years with QTc on screening electrocardiogram (ECG) ≤450 ms. Exclusion criteria included low serum potassium or magnesium and history of cardiac conditions with QT prolongation risk. Bendamustine 90 mg/m2 was given on days 1 and 2 and rituximab 375 mg/m2 on day 1 of each 28-day cycle. On day 2 of cycle 1, ECGs and blood sampling were performed ~15 min prior to bendamustine infusion (baseline), at the end of infusion (EOI)/Cmax, and 1 h post-EOI. The primary measure was change in QTcF at EOI. Secondary measures included outlier analyses of QTcF intervals, pharmacokinetic/pharmacodynamic (PK/PD) analysis, and changes in ECG morphology. The ECG analysis included patients with ≥1 ECG at baseline and on treatment. Results: There were 53 enrolled patients (92% white; 60% male; mean age 63.0 y, range 37-84 y; mean body surface area 2.0 m2, range 1.4-2.6 m2). No clinically relevant signal of any bendamustine effect on cardiac repolarization was detected based on the mean change in QTcF from baseline and outlier analyses (Table). In the PK/PD analyses, predicted change in QTcF for bendamustine and its metabolites, M3 and M4, were not significant: 5.4 ms (upper CI 7.6 ms), 6.0 ms (8.8 ms), and 7.1 ms (10.1 ms), respectively, but there were few data. No new ECG morphologic changes were seen. Conclusions: In this small population with previously untreated advanced NHL or MCL, bendamustine was associated with a small increase in mean QTcF that was not clinically meaningful. Support: Teva BPP R&D, Inc. Clinical trial information: NCT01073163. [Table: see text]
Phase II study of low-dose methotrexate to reduce the incidence of human anti-mouse antibodies in patients receiving I-131 tositumomab as first-line treatment for follicular lymphoma.
Authors: Lebovic Daniel , Avram Anca M , Dewaraja Yuni , Estes Judith , Jacobi Kathryn , Goeman Sara , Kyle Sarah , Chapman Erica , Griffith Kent A. , Kaminski Mark Stefan
Published: March 2020

e19519

Background: I-131 tositumomab is a radiolabeled murine anti-CD20 antibody. In a frontline phase II follicular lymphoma (FL) trial, treatment with I-131 tositumomab produced a 95% ORR with 75% CR (NEJM 352:441, 2005). Median PFS was 6.1 yrs. However, 30% of patients (pts) developed high levels of human anti-mouse antibodies (HAMA) and a serum sickness syndrome within 7 weeks of treatment (early onset HAMA). The 5-yr PFS for this group was 35% compared to 70% for the others (p = 0.003). Low-dose oral methotrexate (mtx) reduces the incidence of human anti-chimeric antibody (HACA) formation in rheumatoid arthritis pts receiving infliximab. We hypothesized that low-dose mtx would likewise reduce the incidence of early onset HAMA when given in combination with I-131 tositumomab. Methods: This is a single arm, phase II study in pts with FL, grade 1-2, stage III or IV disease without limits on marrow involvement, and ≥ 1 of the GELF criteria. Prior therapy, other than focal radiotherapy, is not allowed. Pts must have adequate hematologic, hepatic and renal function, and serum HAMA levels ≤ ULN. I-131 tositumomab is given as a dosimetric dose followed within 1-2 weeks by a therapeutic dose of 75cGy of total body radiation. Pts take mtx 7.5 mg orally once weekly for 3 weeks prior to their dosimetric dose, followed by 7 additional weekly doses. The primary endpoint is the rate of HAMA formation within 7 wks of I-131 tositumomab treatment. Secondary endpoints are overall HAMA incidence, ORR, CR, PFS OS, and safety. Results: Between 8/2011 and 10/2012, 15 pts were enrolled. Median age was 56 (39-83); 11 were male. Three pts had serum HAMA levels >ULN at baseline. One pt had a spontaneous remission prior to starting treatment, leaving 11 evaluable pts. One pt developed early onset HAMA (9%). ORR = 82%, CR = 64%. Two pts progressed with DLBCL. Grade 3 or 4 toxicities included thrombocytopenia n = 3, neutropenia n = 2 and pain n = 3. Conclusions: These preliminary results show that concurrent low-dose oral mtx appears to reduce the incidence of early onset HAMA in FL patients receiving first- line I-131 tositumomab (9% vs. 30% in historical controls) while preserving efficacy. Clinical trial information: NCT01389076.
Sphingolipids as a novel target for the treatment of multiple myeloma.
Authors: Kang Yubin , Kummetha Venketa Jagadish
Published: March 2020

e19534

Background: Multiple myeloma (MM) is the second most common hematological malignancy in the United States and accounts for ~10,600 deaths annually. MM remains an incurable disease and almost all patients will eventually relapse and become refractory to currently available therapeutic agents. There is an unmet need for better understanding the disease’s molecular pathways and for identifying novel therapeutic targets. Sphingolipid metabolism is being increasingly recognized as a key pathway in tumor cell proliferation and in tumor sensitivity to anticancer drugs. We hypothesize that altered sphingolipid metabolism plays an important role in the pathogenesis of MM, thus providing a novel target in the treatment of MM. Methods: We first assayed sphingolipid metabolism including sphingolipid metabolites and sphingolipid metabolizing genes in myeloma cell lines, in freshly isolated human primary CD138+myeloma cells, and in publically available dataset. We then tested the efficacy of the selective SK2 inhibitor (ABC294640) and the SK2 shRNA in killing myeloma cells in vitro. Results: 1) Compared to immortalized B cells, the levels of pro-apoptotic ceramides were decreased whereas the proliferative sphingosine 1-phosphate (S1P) was increased in myeloma cell lines. 2) The expression of several key sphingolipid-metabolizing genes including sphingosine kinase (SK) 1 and 2 was altered in freshly isolated human primary bone marrow myeloma cells and in publically available microarray dataset. 3) The selective SK2 inhibitor (ABC294640) induces apoptotic cell death and inhibits myeloma cell growth with an IC50of ~20 μM in 9 myeloma cell lines. 4) Interestingly, OPM-1 myeloma cell line was extremely sensitive to ABC294640 with an IC50of <5 µM whereas U266 myeloma cell line was resistant to ABC294640. SK2 shRNA induced apoptotic cell death in OPM-1, but not in U266 cells. We are currently investigating the molecular mechanisms underlying the resistance of U266 myeloma cells to ABC294640. Conclusions: Our data demonstrated that sphingolipid metabolism provides an attractive target in the treatment of refractory/relapased multiple myeloma.
Pazopanib in soft tissue sarcoma: Experience from a U.K. expanded-access program.
Authors: Frezza Anna Maria , Benson Charlotte , Miah Aisha , Linch Mark David , Alvarado Rolyn , Mitchell Scott , Esler Claire , Hatcher Helen , Judson Ian Robert
Published: March 2020

e21516

Background: Pazopanib has been recently proven to be a valuable treatment option for patients with metastatic non-adipocytic soft-tissue sarcoma (STS). We present our expanded-access experience to assess the safety and efficacy in routine clinical practice. Methods: Twenty-two patients were treated at 3 UK Sarcoma Centres between December 2011 and September 2012. Response was assessed according to RECIST 1.1 criteria and treatment toxicity graded. Results: Patients had a median age of 50 years (range-21-83), female (54%), 19 (86%) were metastatic. Primary sites were limb (27%), abdominal cavity (46%) and trunk (27%). Median number of previous chemotherapy lines was 2 and all patients were angiogenesis inhibitor naïve. Eighteen patients (82%) started pazopanib at 800 mg daily, 4 (18%) at 600 mg due to borderline PS. The most common side effects included fatigue (G1/2: 59%; G3/4: 9%), hypertension (G1/2: 5%, G3/4: 23%), liver function abnormalities (G1/2: 14%, G3/4: 9%) and diarrhoea (G1/2: 18%). Nine patients (41%) required a dose reduction and four (18%) discontinued the drug due to toxicity. One patient with metastatic pleomorphic sarcoma underwent RFA of the largest lung lesions and was started on pazopanib two weeks later. Despite achieving a partial response, pazopanib was discontinued due to the development of lung cavitation at the site of a previous RFA. Twenty-one patients were available for response evaluation. There were 7 (33%) partial responses (2 synovial sarcoma, 2 undifferentiated sarcoma, 1 fibromyxoid sarcoma, 1 fibromatosis and a 1 retroperitoneal leiomyosarcoma), 5 (24%) stable disease and 9 (43%) progressive disease. Clinical benefit rate (PR+SD) was 56%. Follow-up data were available for 18 patients. At a median follow-up of 25 weeks (range: 9-49), 8 patients (44%) were still alive and 5 (28%) were progression free. Median TTP was 11 weeks (95% CI 5.6-16.3). Conclusions: Pazopanib is effective in the treatment of STS progressing despite standard chemotherapy and has a manageable toxicity profile. However, due to its anti-angiogenic effect, it can interfere with the healing process and its use should be carefully considered in patients undergoing invasive procedures.
Tumor response assessment in locally unresectable or metastatic soft tissue sarcoma (aSTS) patients (pts): A three-year Regina Elena Cancer Institute (IRE) experience with trabectedin therapy.
Authors: Nuzzo Carmen , Anelli Vincenzo , Carpano Silvia , Torsello Angela , Sergi Domenico , Covello Renato , Zoccali Carmine , Vaccaro Vanja , Di Filippo Franco , Giannarelli Diana , Ferraresi Virginia , Cognetti Francesco
Published: March 2020

e21517

Background: Trabectedin has emerged as an effective agent for aSTS pts pretreated with anthracyclines and ifosfamide. Recent data suggest that RECIST criteria could underestimate tumor response. Methods: ASTS pts who received trabectedin as 2nd or further line of treatment were retrospectively reviewed. Tumor response was assessed using RECIST, mChoi and MASS criteria. A good response according to mChoi Criteria was defined as ≥ 10% mean decrease in tumor size or a ≥ 15% decrease in tumor attenuation on contrast-enhanced CT images. Predominantly solid enhancing target lesions were evaluated for marked central necrosis or marked decreased attenuation on axial contrast-enhanced CT images by MASS Criteria. Results: From May 2009 to December 2012, 30 pts with aSTS received trabectedin; pts characteristics are summarized in the table. Median number of previous treatment lines was 2 (range 1-5); median number of cycles administered was 4 (range 1–22 cycles). Treatment was well tolerated; main causes of dose-reduction were hematological and liver toxicities. Median progression free survival (PFS) was 8 wks (95% CI: 2-14); median overall survival (OS) was 44 wks (95% CI: 18-94). According to RECIST, 4 pts (15.4%)obtained a partial response (PR)), 9 pts (34.6%) achieved a stable disease (SD); tumor control rate was 50%. 13 pts (43%) obtained a progression of disease (PD). CT Scans of 13 pts were available for comparison of different response assessment methods (2 RP, 5 SD, and 6 PD). In our small series of pts concordance between the three radiologic methods tested was 90%. Conclusions: In aSTS patients treated with trabectedin, there is substantial concordance between mChoi, MASS and RECIST criteria. Prospective validation of the potential role of mChoi and MASS criteria in this setting is warranted. [Table: see text]
Relationship of the oncogene Notch and CXCR4/SDF1 signaling in human epithelial ovarian cancer.
Authors: Konala Venu , Platonova Natalia , Colombo Michela , Mirandola Leonardo , Lazzari Elisa , De Simone Daria , Radhi Saba , Aulakh Amardeep , Zepeda Kristopher , Wade Raymond , Mer Jesse , Saadeh Charles , Yu Yuefei , Jenkins Marjorie , Figueroa Jose A. , Cobos Everardo , Chiriva-Internati Maurizio , Chiaramonte Raffaella
Published: March 2020

e22003

Background: Ovarian cancer (OC) is the most aggressive gynecologic cancer. Understanding OC molecular pathogenesis is critical to provide novel therapeutic strategies. We aim to elucidate Notch oncogenic role in OC by focusing on its extensive crosstalk with other important pathways as CXCR4/SDF1alpha chemokine system whose involvement in OC development and metastasis is well recognized. Methods: We used flow-cytometry, cell cycle analysis, real-time PCR, and Transwell chemotaxis assay to investigate the outcome of Notch signaling withdrawal on tumor cell response to CXCR4 and SDF1alpha. Results: The analyzed OC cell lines expressed high levels of CXCR4 and its ligand SDF1alpha. Treatment with DAPT, an inhibitor of Notch activity, reduced OC cell proliferation and blocked cell cycle in G0/G1 phase without affecting apoptosis. In addition, Notch withdrawal diminished CXCR4 and SDF1alpha expression levels and hampered SDF1-driven migration and proliferation. Conclusions: Notch deregulation might affect important features of OC such as cell growth and migration through the modulation of CXCR4/SDF1a pathway. This indicates that these intertwined pathways are promising therapeutic targets in OC.
Effect of pharmacologic inhibition of c-Jun N-terminal kinase (JNK) signaling pathway on cell proliferation and cell cycle progression in human granulosa cell tumor.
Authors: Oktem Ozgur , Muftuoglu Meltem , Senbabaoglu Filiz , Urman Bulent
Published: March 2020

e22004

Background: No data are available regarding the signaling pathways that controls the proliferation of granulosa cell tumors (GCT). Preliminary findings showing the activation of c-Jun N-terminal kinase (JNK) signaling pathway in the proliferating granulosa cells has led us to investigate the role of this pathway in human GCT. Methods: Human GCT line COV 434 was used. Cell proliferation was monitored real-time quantitatively for 120h using an impedance-based system. Two different pharmacologic JNK inhibitors SP600125 and AS601245 were used. Their inhibitory concentrations were determined in western blot. Cell cycle was analyzed with flow cytometry and apoptosis with yo-pro-1 staining. Results: First, the growth characteristics of this cell line was delineated (Table 1A). Then the cells were treated with the inhibitors at the indicated doses during the log phase. Their proliferation was significantly halted in a dose-dependent manner by both inhibitors (Table 1B). Furthermore, the cells failed to complete mitosis, and began to accumulate at G2 in a dose dependent manner when JNK pathway was interrupted with AS601245 (59%) and SP600125 (39%) during G2/M transition compared to control cells (7%) proceeding through G2/M phase regularly (p<0.001). Compared to 3.5% of control cells, 14% and 30% of the cells underwent apoptosis when treated with 50 µM SP600125 and AS601245, respectively. At 100 µM, the apoptotic fraction increased to 68% and 76%, respectively (p<0.01). Conclusions: These results suggest that pharmacologic manipulation of JNK pathway may provide a therapeutic benefit in the treatment of GCT for which currently, no curative therapy exists beyond surgery. Funded by a Grant to Ozgur Oktem (TUBITAK109S164). [Table: see text]
Malignancy testing by AmpliSeq cancer panel sequencing-based assessment in NSCLC.
Authors: Lai Rensheng , Wu Xiaobin , Sun Yi , Chen Jie , Xie Ling , Yu Bo , Sun Lele
Published: March 2020

e22026

Background: Bringing PGM sequencing into clinical laboratories needs the same confidence and consistency with Sanger sequencing. We have developed a approach to assess the known EGFR mutation of NSCLC using AmpliSeq Cancer Panel and demonstrated high specificity and sensitivity. Methods: Fifteen standard cell line samples with known EGFR mutation frequency (1%, 5%, 20%, 50%) were issued by CPC of National Health Ministry. These were first examined by sanger sequencing, then by AmpliSeq cancer panel on PGM. Results were compared each other. Next, we applied the AmpliSeq cancer panel sequencing to test the mutation status of 8 clinical malignancy FFPE samples including 4 NSCLC, 1 Esophagus Cancer, 1 Breast Cancer, 1 Gastric Cancer and 1 stroma tumor. gDNA was extracted from FFPE section then subject to library construction. Emulsion PCR was performed and beads were enriched, Sequencing was performed with 200 bp readlength by pooling 2-4 samples on one 314 chip. Data yield ranges from 13,311 to 203,061 eligible reads (2 Mb to 44M) per sample. Results: Validation of AmpliSeq Cancer Panel Sequencing: Among all 15 samples, 14 cases (14/15, 93%) were confirmed by Sanger sequencing. AmpliSeq FFPE malignancy: For all 8 FFPE samples, the sequencing depth of EGFR site ranges from 20x to 2000x with 100% on-target coverage. Each patient can be identified with more than 3 target drugs based on mutant frequency (>5%) with 99% confidence (CI 0.99-0.995). All patients were shown with the presence of rare subpopulations of interrogating 739 mutations in 46 genes (0.1% to 94% allele frequency). Conclusions: AmpliSeq cancer panel sequencing shows equal or higher sensitivity to sanger sequencing . Call analysis shows that a minimum of 100x coverage is required to detect mutations ≥ 5%. [Table: see text]
CARD8/BCL-2 cascade in early adaptive drug resistant tumor escape against targeted inhibitors in NSCLC.
Authors: Bagai Rakesh K. , Zhang Wei , Leahy Patrick , Yin Lihong , Ma Patrick C.
Published: March 2020

e22032

Background: Lung cancer targeted therapy is largely limited by inevitable recurrent resistant disease after initial response to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), typically accompanied with divergent late acquired resistance mechanisms. We now focused on studying the emergence of early adaptive resistance to uncover attractive therapeutic targets to overcome drug resistance. Methods: HCC827 cells were treated with EGFR-TKI (0-9 days) with apoptosis pathway-specific QPCR array and TLVM analysis performed. MTS and crystal violet assays were performed. Western blot analysis was performed to examine prosurvival signaling developed against erlotinib, alone or in combination with MET inhibitor SU11274. IHC was performed on lung cancer tumor microarray (TMA) using BCL-2 and caspase-recruitment domain-containing protein 8 (CARD8) antibodies and graded (4 tier scoring system). NSCLC cell lines and murine xenograft models (HCC827, H1975) were developed for resistance biomarkers expression analysis in pre-/post-TKI treatment using anti-human CARD8, p-STAT3 and BCL-2 antibodies. Results: We characterized the emergence of early resistant lung cancer cells in escape against targeted TKIs with 100-fold higher IC50 in adaptive drug resistance. The resistant cells that evaded EGFR-TKI based targeted inhibition exhibited MET-independent induction of CARD8 and STAT3/BCL-2 mitochondrial prosurvival signaling in cellular quiescence, and inhibited cytoskeletal functions. Expression analysis studies demonstrated common tumor-associated expression of CARD8 but relatively low BCL-2 level in NSCLC. In vitro cell line studies suggest that CARD8 induction was preceded by a resurgence of STAT3 activation. In vivo xenograft model (HCC827/erlotinib; H1975/erlotinib+ SU11274) also verified upregulated CARD8/BCL-2 activation within early resistant cells. Conclusions: Resistant tumor cells that evaded EGFR inhibitors, alone or in combination with MET inhibitors, exhibited increased expression of CARD8-STAT3/BCL-2 prosurvival signaling cascade. Further studies to define the mechanism of CARD8 in promoting adaptive tumor drug resistance would be warranted.
Oncogenic mutations in colorectal cancer, indications for anatomical sites, and targeted intervention.
Authors: Corso Giovanni , Pascale Valeria , Flauti Giuseppe , Marrelli Daniele , Roviello Franco
Published: March 2020

e22037

Background: Oncogenic mutations, such as KRAS, in colorectal cancer patients are considered standard molecular biomarkers that predict the clinical benefit for the targeted intervention with EGFR inhibitors. In addition, these mutations are associated with specific anatomical area in the colon tumor development, as BRAF mutations with the microsatellite instability. Methods: In this translational study we aim to assess the mutation frequencies of the EGFR [hotspot area and polyadenine deletions (A13_del)], KRAS, BRAFV600E, and PIK3CA oncogenes in a series of 280 colorectal cancer patients. Microsatellite instability phenotype is considered in this series. All patients' clinicopathological data were considered for statistical analysis and associations. Results: In this study, we verified multiple associations between oncogenic mutations and specified clinicopathological tumor features. Respectively, we identified the following significant results: 1) EGFR A13_deletions are associated with right colon carcinoma (22.2% vs. 3.3%; p<0.005), mucinous histotype (16% vs. 7.8%; p=0.042), G3 grading (19% vs. 7.3%; p=0.024) and microsatellite instability status (p<0.005); 2) PIK3CA mutations are related mucinous histotype (12% vs. 4.4%; p=0.021) 3) KRASG12 and KRASG13mutations are correlated respectively with the left (91.4% vs. 59.3%) and right (40.7% vs. 8.6%) colon cancer development (p<0.005), and finally 4) microsatellite instability is associated with right colon tumors (28.4% vs. 5.5%; p<0.005). Conclusions: Mostly, we verified a high frequency rate of the KRASG13 and EGFR A13_del oncogene mutations in right colon cancer; whereas KRASG12 codon mutation occurs more frequently in left colon cancers. In particular, we assessed that right colon cancer is associated with specific molecular characteristics, in comparison to left colon tumors. These evidences, in association with specific clinicopathological data, can delineate novel approaches for the colorectal cancer classification and targeted intervention.
Effect of triptolide on gemcitabine-mediated apoptosis in pancreatic cancer cell lines.
Authors: Wang Wei , Lin Wenjie
Published: March 2020

e22043

Background: Pancreatic cancer is notorious for its resistance to anticancer drug. gemcitabine, the most effective anticancer drug currently available for palliative chemotherapy in pancreatic cancer patients, hardly achieves clinically satisfactory response rates. Therefore, it is urgent to develop novel agents that overcome drug resistance of pancreatic cancer. In this study, we demonstrated for the first time that triptolide, a diterpenoid triepoxide purified from the Chinese herb Tripterygium wilfordii Hook F, can enhance gemcitabine-mediated apoptosis in human pancreatic cancer cell lines (AsPC-1 and PANC-1). Methods: Pancreatic cells were treated with gemcitabine or triptolide alone, and combination of them, cell viability was measured by MTT, and apoptotic cells were assessed by flow cytometry for Annexin V/PI staining and western blot for cleaved caspase-3, 8, 9 and PARP. To explore the mechanism of enhancement of triptolide and gemcitabine, proteomic approach was used to screen the related apoptosis factors. Results: Triptolide demonstrated toxicity on pancreatic cancer cell lines with the IC50 of 25-40 nM. The combination of triptolide (IC30 concentration) and gemcitabine enhanced the cytotoxicity significantly compared to gemcitabine alone. Combination index (CI) indicated the effect of triptolide plus gemcitabine was synergistic. Furthermore, pancreatic cancer cells treated with triptolide plus gemcitabine exhibited increased apoptosis, as evidenced by stronger Annexin V/ PI staining, higher levels of pro-apoptotic proteins including cleaved caspases and activated PARP compared to cells treated with gemcitabine alone. In the mechanism study, vimentin, a mesenchymal marker, was screened out to be correlated with enhanced apoptosis induced by triptolide plus gemcitabine. Conclusions: 1) Triptolide has a potent therapeutic effect on pancreatic cancer cell lines; 2) the combination of triptolide with gemcitabine enhances the apoptosis in vitro, low concentration of TPL showed synergistic effect with gemcitabine; 3) The enhanced apoptosis induced by triptolide plus gemcitabine is correlated with vimentin expression inhibited by triptolide.
The prognostic impact of high MET gene copy number on non-small cell lung cancer: A meta-analysis of eight studies.
Authors: Dimou Anastasios , Non Lemuel , Syrigos Kostas N
Published: March 2020

e22049

Background: The risk stratification of NSCLC on the basis of molecular information is a key approach in the clinical management of patients with the disease. MET is the receptor for HGF and is present in the membrane of NSCLC cells. The most common genetic alteration associated with MET is the gene amplification. Methods: Two independent investigators applied parallel search strategies with the terms "MET" AND "lung cancer", "MET" AND "NSCLC", "met gene copy number" AND "prognosis" in PubMed through November 2012. We selected the studies that investigated the association of MET gene copy number with prognosis. A quality score that assessed the lab methods, the generalizability and the analysis, was assigned to each study that was finally included in the analysis. Results: Among 791 studies that were identified in the initial search, we retrieved 8 cross sectional studies on retrospective cohorts with adequate data regarding the prognostic impact of MET gene copy number on the survival of patients with NSCLC. Among the studies, 5 used FISH and the remaining 3 used RT PCR to assess the MET gene copy number in the primary tumor. All 8 studies used tissue from surgically resected specimens. MET gene copy number predicted worse overall survival when all studies were combined in a fixed effects model (HR=1.35, 95% CI 1.17-1.57). We calculated the I2 statistic to assess heterogeneity (I2=44%, p=0.09). There were four studies where a higher gene copy number predicted a better outcome and four in which the opposite was true. The Egger’s regression intercept showed no significant publication bias (p=0.38). Conclusions: Higher MET gene copy number in the primary tumor at the time of diagnosis predicts worse outcome in patients with NSCLC.