bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of epigenetic aberrant promoter methylation in serum DNA for detection of pneumoconiosis-associated lung cancer
Authors: Umemura S. , Fujiwara K. , Tabata M. , Kishimoto T. , Gemba K. , Kodani T. , Takigawa N. , Kiura K. , Ueoka H. , Tanimoto M.
Published: March 2020

20059

Background: Pneumoconiosis is known to be associated with an independent increased risk of lung cancer. However, it is difficult to detect lung cancer radiographically in patient with pneumoconiosis because of pre-existing diffuse pulmonary shadows. The purpose of this study is to evaluate the usefulness of serum DNA methylation for detection of pneumoconiosis-associated lung cancer. Methods: We investigated to identify promoter methylation status of RAR-beta, p16INK4a, MGMT, DAPK, and RASSFIA gene in serum DNA using methylation-specific PCR. Sera were obtained from 31 control patients with non-malignant-non-pneumoconiotic pulmonary disease (Group I), 36 with pneumoconiosis (Group II), and 5 with pneumoconiosis-associated lung cancer (Group III). DNA was extracted using QIAamp DNA Blood Midi Kit (Qiagen, CA). Following DNA bisulfite modification using CpGenome DNA Modification Kit (Intergen, NY), PCR was performed with primers for methylated or unmethylated promoter sequences. Results: Median (range) age of patients were 61 (26–78), 71 (49–86), and 69 (56–78) for Group I, II, and III respectively. Median (range) duration of silica and asbestos exposure were 33 (3–47) years and 33(32–40) years for Group II and III respectively. All of 5 pneumoconiosis-associated lung cancer were adenocarcinoma. Eleven patients (35.5%) in Group I, 19 patients (52.8%) in Group II, and 5 patients (100%) in Group III were shown to have methylation of at least one gene. The total number of methylated genes per patient were, 0.35, 0.69, and 1.20 for groups I, II, and III respectively (p = 0.013, Kruskal-Wallis analysis). In Group II, methylation status did not correlate with duration of occupational exposure, smoking history, radiographic findings, and age. Conclusions: In patients with pneumoconiosis, monitoring of aberrant promoter methylation of serum DNA might be useful for assessing the risk of lung cancer and early detection of lung cancer.

No significant financial relationships to disclose.
Low-level amplifications of the human telomerase gene (TERC) regularly is associated with vulvar intraepithelial neoplasia
Authors: Gebauer G. , Schleibaum J. , Aulmann S. , Schirmacher P. , Sohn C. , Sinn P.
Published: March 2020

20060

Background: Invasive squamous carcinomas of the vulva frequently harbor gains on the long arm of chromosome 3, including the human telomerase gene locus (TERC). Vulvar cancer develops from precursor lesions known as vulvar intraepithelial neoplasia (VIN) which frequently occur simultaneously in multiple locations and have a considerable risk or recurrence during follow-up. We investigated TERC gene copy number in a series of vulvar intraepithelial neoplasias to determine the risk of progression in these lesions. Methods: A total of 20 cases of high grade vulvar intraepithelial neoplasias (VIN 3) were analyzed using a two-color fluorescence in situ hybridization (FISH) assay detecting the human telomerase gene locus (3q26) on paraffin embedded tissue sections. Two samples were excluded for technical reasons. Results: In 11 cases, gains of chromosomal material at 3q26 were detected with an average of 1.48 copies per chromosome. In another patient a high level amplification was found. The remaining two cases, four centromere and 3q26 hybridization signals were observed, consistent with a tetraploidy of chromosome 3. No changes in DNA copy number on chromosome 3q were detected only in four lesions. Conclusions: Our findings demonstrate high frequency of gains on chromosome 3q in the majority of high grade vulvar lesions. This suggests an involvement of the human telomerase gene locus in the pathogenesis of vulvar cancer.

No significant financial relationships to disclose.
Effect of antioxidants on STAT3, cell survival gene products, apoptosis and chemoresistance in lung cancer cells
Authors: Pathak A. K. , Bhutani M. , Pathak R. , Mohan A.
Published: March 2020

20067

Background: We had earlier demonstrated that antioxidants (beta carotene, vitamins C and E) enhance the apoptotic effect of paclitaxel and carboplatin in vitro and there was a suggestion that when used as an adjunct to the above chemotherapeutic agents in patients with NSCLC, they might enhance the efficacy of the latter. To elucidate the mechanism underlying the synergistic effect between antioxidants and chemotherapy, in the present study we evaluated the effect of antioxidants on STAT3 activation in NSCLC cells. Since this transcription factor has been shown to regulate multiple oncogenic pathways, including pathways regulating tumor cell survival, we further studied how STAT3 modulation relates to apoptosis and downstream apoptosis regulatory proteins. Methods: H520 cells were treated with antioxidants. STAT3 proteins were assessed by western blotting and their nuclear localization was evaluated by immunocytochemistry. Antiapoptotic proteins (survivin, Bcl-2, Bcl-xl and cyclin D1) were evaluated by Western blotting. To assess the interaction, cells were treated with chemotherapy (Paclitaxel and Carboplatin) with or without antioxidants and the effect was assessed by Live & Dead assay, MTT assay and flowcytometry. Results: Antioxidants inhibited STAT3 phosphorylation in a dose and time dependent manner before the initiation of cell death and depleted the nuclear pool of STAT3. This effect was specific since there was no effect on STAT5 phosphorylation and was reversible since STAT 3 reverted back to its constitutively activated state after the removal of antioxidants. Antioxidants suppressed the expression of antiapoptotic proteins Bcl -2, Bcl-xl, cyclin D1 and survivin. This resulted in the inhibition of proliferation of cells and induction of apoptosis as evident on caspase 3 activation and PARP cleavage. Antioxidants arrested the cells in G0/G1 phase of cell cycle making them more vulnerable to the cytotoxic effects of chemotherapy. This synergy was evident on live and dead assay. Conclusions: Our results demonstrate that antioxidants are potent inhibitors of STAT3 phosphorylation and can have a potential role as an adjunct to chemotherapy in lung cancer.

No significant financial relationships to disclose.
Combined expression of bcl-2, bcl-6, HIF, HGF, c-kit, neu/cerb2 and VEGF-C and their clinical significance in laryngeal squamous cell carcinoma
Authors: Kara I. O. , Uguz A. , Sahin B. , Kilic-Bagir E. , Erkisi M.
Published: March 2020

20079

Background: Proteins regulating the cell cycle and cell death are frequently abnormally expressed in cancer. Several of these, particularly Bcl-2, Bcl-6, neu/cerb2 have been widely suggested as possible prognostic markers in diverse human malignancies. Their role in predicting outcome in squamous cell carcinomas of the head and neck is unclear. In addition c-kit protein has been shown in tumorogenesis. Because HNSCC preferentially metastasizes to regional lymph nodes, we investigated the expression of bcl-2, bcl-6, HIF, HGF, c-kit, neu/cerb2 and VEGF-C and their clinical significance in laryngeal squamous cell carcinomas by semiquantitative immunohistochemistry. Methods: We studied 115 patients with stage I o III tumors, all were treated with surgery ± postoperative irradiation/chemotherapy by a single institute. We studied the patients retrospectively to test the association between expression of Bcl-2, bcl-6, HIF, HGF, c-kit, neu/cerb2 and VEGF-C, as assessed by immunohistochemistry in formalin-fixed paraffin-embedded tissue and evaluated by two pathologist. We scored the expression of the proteins from negativ to severe expression. In addition we evaluated the degree of tumor grade, necrosis and also inflamation, respectively. Results: Within 115 patients we found severe expression of bcl-2 in 2 (1%), bcl-6 in 16 (13%), HIF in 57 (49%), HGF in 38 (33%), c-kit in 4 (3%), neu/cerb2 in 17 (14%) and VEGF-C in 11 (9%) patients, respectively. We found significant correlation between bcl-2 and necrosis (p = 0.003), HGF and inflamation (p = 0.05), c-kit and necrosis (p = 0.04), c-kit and tumor grade (p = 0.03) respectively. We found that the significant relation between bcl-6 and HIF (p = 0.000), bcl-6 and HGF (p = 0.02), bcl-6 and c-kit (p = 0.005), bcl-6 and VEGF-C (p = 0.000), HGF and VEGF-C (p = 0.005) respectively. Also we found that the significant correlation between tumor grade and HGF (p = 0.05). Conclusion: These data indicate that assessing expression of bcl-2 or bcl-6, c-kit and HGF is unlikely to be prognostically useful for surgically treated laryngeal carcinoma.

No significant financial relationships to disclose.
Analysis of gemcitabine pathway genotypes in ethnic Asians and their relationship with outcome in non-small cell lung cancer (NSCLC) patients
Authors: Soo R. A. , Lee S. C. , Ng S. S. , Chong P. Y. , Tham L. S. , Wang L. Z. , Lee H. S. , Soong R. , Goh B. C.
Published: March 2020

2008

Background: Genotypic variation for drug metabolising enzymes, targets and transporters is associated with inter-patient and inter-ethnic variability in toxicity and efficacy. Gemcitabine is a nucleoside anti-metabolite used in a range of solid tumors. Aims: 1) to determine genotype distribution of genes involved in gemcitabine pharmacology pathway in ethnic groups; 2) to evaluate the association between genotypes and treatment related outcomes in patients with chemo-naive, advanced NSCLC receiving gemcitabine based chemotherapy on a prospective study. Methods: Candidate genes involved in gemcitabine pharmacology were identified from a comprehensive search of public databases (NCBI, PharmGKB), and publications. 25 variants of 9 candidate genes (RRM-1, SLC28A1, SLC28A2, SLC28A3, SLC29A2, POLA2, DCTD, CDA, TS) were genotyped from blood in 94 healthy donors and 76 cancer patients (breast 22, NSCLC 54) with pyrosequencing. Chi-squared, Kruskall-Wallis, and Kaplan-Meier analysis were used to evaluate associations between genotypes and ethnic groups and clinical end points. Results: Significant differences in genotype distribution between Chinese, Malay and Indian were seen in 5/25 loci (see table ). In NSCLC patients, POLA2 2089 (G > A) was associated with neutropenia (p = 0.022), SLC28A1-1576 (C > T) with neutropenia nadir (p = 0.030) and thrombocytopenia nadir (p = 0.037), RRM-1 -524 (T > C) with neutropenia (p = 0.024) and SLC28A2–283 (CA 18.3 vs CC 8.5 months, p = 0.004), and SLC28A2–22 (CC 8.0 vs CT 18.3 months, p = 0.002) with survival. Conclusions: Our results suggest there is significant genotypic variability among ethnic groups. Variants in gemcitabine transporters and targets may be useful indicators of gemcitabine related toxicity and survival. Further studies should be performed to confirm these preliminary findings.

[Table: see text]

No significant financial relationships to disclose.
Retrospective analysis of cutaneous T-cell lymphoma (CTCL) biopsy samples for interleukin-2 receptor (IL2-R) subunit expression by immunohistochemistry (IHC) and real-time quantitative PCR (RT-PCR): Correlation to response to denileukin diftitox
Authors: Alexander V. J. , Bissonnette R. P. , Rosen J. I. , Cooke T. , Foss F. M.
Published: March 2020

20080

Background: Denileukin diftitox, a chimeric diphtheria toxin-IL2 protein, exerts its cytotoxic effect on cells expressing high or intermediate affinity IL2-R. IL2-R is a complex of 3 distinct polypeptide chains: IL2-Rα (CD25), IL2-Rβ (CD122), and IL2-Rγ (CD132). IL2-R α/β/γ heterotrimer binds with high affinity, IL2-R β/γ with intermediate affinity, and IL2-R α/γ with low affinity. Denileukin diftitox is indicated for patients with persistent or recurrent CTCL whose malignant cells express IL2-Rα protein. There is no currently available methodology to measure expression of the high affinity receptor in patient samples. This study was initiated to discover a correlation between expression of IL2-R components (at the mRNA and protein levels) and clinical response to denileukin diftitox. Methods: Of 33 available frozen clinical CTCL biopsy samples, 27 (10 responders) were sufficient for IHC. IL2-Rβ staining intensity was assigned a value of 0 (no staining relative to background), 1+ (weak), 2+ (moderate), or 3+ (strong). 22 samples (8 responders) were sufficient for RT-PCR. mRNA levels of the 3 IL2-R chains were normalized to 36B4 mRNA or 18S rRNA. Correlation between clinical response and IL2-R chain expression was assessed using Spotfire Decision Site. Results: The population of samples tested was representative of the study population. 26 of 27 samples exhibited 2+, all exhibited 1+, and none 3+ IL2-Rβ staining. The percentage of cells stained varied from 20%–90% for 1+ and 10%–80% for 2+ with no correlation to response. Correlation of levels of individual IL2-R subunit mRNA expression with response could not be established. Conclusions: Neither the IL2-Rβ IHC nor the RT-PCR assay, at current sensitivity levels, appear to identify denileukin diftitox responders. Larger sample datasets from current studies, and further assay refinement, are ongoing to define the merits of these approaches.

[Table: see text]
Neuroendocrine cancer of the kidney: Clinical implications of expression microarray analysis
Authors: Harris W. B. , Assikis V. , Yin-Goen Q. , Lewis M. M. , Lawson D. , Amin M. B. , Young A. N. , Petros J. A.
Published: March 2020

20093

Background: Conventional clinical trial strategies cannot be used to develop new therapies for rare malignancies. Neuroendocrine neoplasms of the kidney are extremely rare. Here we compare the gene expression profiles of a neuroendocrine tumor from a 32 year old male with a horseshoe kidney, a large renal primary and metastatic thyroid lesions to the profiles of other cancers. Response to systemic therapy is also reported. The ultimate objective of this project is to identify new therapeutic targets for patient-specific cancer therapy. Methods: After informed consent, tissue from the patient was analyzed by cytology and immunohistochemistry. Total RNA was isolated from formalin-fixed paraffin-embedded archival reference tissues with Arcturus Paradise Reagents. Gene expression was analyzed with Affymetrix X3P arrays of the entire expressed genome. A supervised significance analysis of microarrays (SAM) procedure, using 400 data permutations, was performed to identify gene products that were differentially expressed. Clinical outcomes were documented in the patient’s medical record on the basis of standard medical imaging. Results: Cytology, immunohistochemistry and microarray studies revealed similar patterns for the primary and metastatic lesions. Gene expression profiles of the patient’s kidney and thyroid lesions, as well as other kidney and neuroendocrine tumors, identified 11 genes overexpressed significantly in the neuroendocrine tumors. Each was associated with neuroendocrine biology. The patient had a mixed response to chemotherapy with gemcitabine and cisplatin and progressive disease after 3 monthly treatments with sandostatin. He awaits assessment of response to metaiodobenzylguanidine (MIBG). Conclusions: Gene expression profiling was effective in characterizing an unusual case of renal cancer with metastasis to thyroid, though no new therapeutic target has yet been identified. Comparing response or resistance to therapy with microarray analysis may prove to be an effective strategy for developing patient-specific cancer therapy. This work was supported, in part, by a VA Merit Review Award.

No significant financial relationships to disclose.
Effects of polymorphism (plm) in interleukin (IL) 1 and IL10 genes on surgery outcome in ovarian cancer (OC)
Authors: Braicu E. I. , Mustea A. , Koensgen D. , Pirvulescu C. , Sun P. , Chekerov R. , Nuernberg P. , Toliat M. R. , Lichtenegger W. , Sehouli J.
Published: March 2020

20094

Background: IL1 and IL10 are overexpressed in 55% to 98% of advanced epithelial OC. Different studies demonstrated IL1 and IL10 as independent prognostic factors for OC. This study was performed to investigate the influence of plms of IL-1A, IL-1B and IL-10 gene on the occurrence tumor biology of OC. Methods: In a prospective, monocentric, case-control study 147 pts with OC (71 pts with primary OC and 77 pts with recurrent OC) and 129 pts with no history of malignancies were genotyped for plms of the IL-1 gene (IL-1A -889 T/C and IL-1B -511 C/T promoter) and IL-10 gene (IL-10 -1082 G/A, -819 C/T and -592 C/A). All pts were collected within Tumor bank Ovarian Cancer Project. Genomic DNA fragments were amplified by PCR and the allele sequences were obtained by pyrosequencing. We analyzed significance using the Chi-Square by Pearson and Fisher’s exact test. Statistical significance was determined by Mann-Whitney test. A two-tailed p value <0.05 was considered statistically significant. Overall survival (OS) was estimated using the Kaplan-Meier product limited method. Results: The median age of the pts with OC was 55.5 years (range 19–81 y). Median follow-up period was: 31.5 months (range: 1–225 mo). There were no significant associations between allelic frequencies for IL-1A and IL-1B in OC. The IL-10 plm in -819 and -592 positions significantly correlates with the postoperative residual tumor (p = 0.036 and p = 0.035 respectively). For pts without any postoperative tumor residuals the most encountered genotype was C/C at −819 and −592. The same genotype was associated in univariate analysis with a better OS (95% CI = 37.5–94.5 and 40.8–91.2, respectively). In the multivariate models there was no correlation between any plms and disease-free and OS. Postoperative residual tumor (OR =0.202, 95% CI = 0.088–0.462, p = 0.0001) and ascites volume (OR = 0.106, 95% CI = 0.046–0.248, p = 0.026) were independent prognostic factors for reduced OS. Conclusions: Plm of IL1A, IL1B, IL10 showed no association with occurrence of OC. IL-10 promoter plm may influence the tumor biology and could potentially predict results of radical surgery.

No significant financial relationships to disclose.
Swedish ovarian cancer patients are found with a high frequency of human leucocyte antigen (HLA) A2, -B7, -B12, -B15 -B44 and -DRB1–4 haplotypes
Authors: Bergfeldt K. , Gamzatova Z. , Tholander B. , Bergdahl M. , Nordström B. , Hjerpe E. , Masucci G.
Published: March 2020

20100

Background: There is an increased interest on the association between HLA markers and prognostic outcome in several cancer forms. We have previously reported that the high prevalence of the A2 allele in Scandinavia, which decreases with latitude in Europe, correlates to a similar decrease in ovarian cancer mortality among European countries. We have also published the association of this allele with severe prognosis in serous adenocarcinoma of the Ovary in stage III-IV. We now present a complete distribution of the HLA- A, -B -C and DRB1 alleles and haplotypes in relation to histology and clinical stage. Methods: An unbiased selection of epithelial ovarian cancer patients (n = 56) recorded by age, histology, stage and treatment were analyzed for HLA-A, -B, -C and -DRB1 genotypes by PCR/sequence-specific oligonucleotide hybridization procedure (PCR/SSOP). HLA frequencies from healthy Swedish bone marrow donors were used as comparison. Results: HLA-A2, -B7, -B15, -B44 as well as the A2, -B15, -DR4 haplotypes frequency are significantly higher than in the healthy Swedish population. We noticed a high concentration of this haplotypes among the serous adenocarcinomas. Seven patients were homozygotes for A2 allele (23%), two times the healthy Swedish population (12%), and 3 times the median frequency in Europe (8%). Six of these had the haplotype -A2, -B7. Conclusions: HLA-A2 homozygotes and some HLA-A2 -B and -DRb1 haplotypes are higher expressed than in healthy individuals. These observations corroborate the relevance of HLA in association with ovarian cancer, previously partially investigated. Ongoing studies consider the relationship of HLA-A2 and its haplotypes to possible oncogenes and prognosis in ovarian cancer.

No significant financial relationships to disclose.
Multicentric descriptive study of the lung cancer treatment
Authors: Quintana M. , Pallares C. , Martinez I. , Gich I. , Bonfill X.
Published: March 2020

20109

Background: Given the insufficient available information on the clinical characteristics and the results of the lung cancer treatment at a population level, we performed a large multicentric descriptive study in the main hospitals in our city. Methods: We analyzed the patients diagnosed and treated for lung cancer during 1999 in five University Hospitals in Barcelona (Spain). We performed a retrospective analysis of the individual data obtained from the clinical records using a specifically validated questionnaire. A descriptive analysis was performed, stratifying for type of tumor, stage, and institution. The follow-up for the complete population was 5 years. Results: We included 836 patients (94.6% male, 5.4% female) with an average age of 64 years. Karnofsky was ≥ 70% in 85% of patients. Histologic distribution was non-small cell (NSCLC) 85.5%, small-cell (SCLC) 14.5%. 42% of cases were stage IV (38% NSCLC, 67.8% SCLC). First treatment was performed in 82% of cases with therapeutic intention (52% was chemotherapy). Pulmonary resection with lobectomy and platinum-based chemotherapy were the more frequent treatments. Complete remission was obtained in 25% of cases. Survival was 42.0, 17.9 and 12.6% at 1, 3 and 5 years respectively (NSCLC: 43.8, 19.4, 13.1%; SCLC: 31.9, 9.5, 9.5%). The mean interval first visit - treatment start was 2 months. The mean of direct costs per patient was 9000 €. The cost per life-year gained was 5200 € for NSCLC and 7600 € for SCLC patients. Conclusions: The obtained results allow to describe the clinical characteristics of a wide cohort of patients diagnosed of lung cancer and their survival to 5 years. Globally, these results are similar to those who have been published in our country and others, and constitute a very thorough referent for assessing the impact of future therapeutic advances. The comparison among hospitals can also be the matter of further analysis for identifying the causes of the observed differences. Equally, specific strategies for some patients’ subgroups could be designed in order to improve the cost-effectiveness of their treatment.

No significant financial relationships to disclose.
Development of a clinically feasible molecular assay to predict recurrence of Dukes’ B colon cancer
Authors: Jiang Y. , Zhang Y. , Briggs T. , Talantov D. , Mazumder A. , Atkins D. , Wang Y. , Deleany C. , Brown M. , Casey G.
Published: March 2020

20110

Background: The 5-year survival rate of Dukes’ B colon cancer patients is approximately 75%. In our earlier genome-wide measurements of gene expression we have identified a 23-gene signature that sub-classifies Dukes’ B tumors and may provide better means of risk assessment on an individual basis for these colon cancer patients. The aim of this study is to validate this gene signature in an independent and more diverse group of patients, and further develop this prognostic signature into a clinical feasible test using formalin-fixed paraffin-embedded (FFPE) tissue samples. Methods: Using Affymetrix U133a GeneChip we analyzed the expression of the 23 genes in total RNA of frozen tumor samples from 123 Dukes’ B patients who did not receive adjuvant systemic treatment. Furthermore, we developed a quantitative RT-PCR assay for this gene signature in order to perform the test with standard clinical FFPE samples. Results: In the independent validation set of 123 patients, the gene signature proved to be informative in identifying patients who would develop distant metastasis (hazard ratio, HR 2.56; 95% confidence interval CI, 1.01–6.48), even when corrected for the traditional prognostic factors in multivariate analysis (HR, 2.73; 95% CI, 0.97–7.73). The RT-PCR assay developed for this gene signature was also validated in an independent set of 114 patients as a strong prognostic factor for the development of distant recurrence (HR, 6.38; 95% CI, 2.88–14.2) in univariate analysis and in multivariate analysis (HR, 13.3; 95% CI, 5.13–34.4). Conclusions: Our data provide not only a validation of the pre-defined prognostic gene signature for Dukes’ B colon cancer patients but also a clear feasibility of testing the gene signature using RT-PCR with standard FFPE specimens. The ability of such a test to identify patients that have an unfavorable outcome demonstrates potential clinical importance that could lead clinicians to choose a more aggressive therapeutic option for the high-risk patients.

[Table: see text]
Effective detection of the epidermal growth factor receptor mutation in pathologic specimens of non-small cell lung cancer patients by a rapid and sensitive detection system, the peptide nucleic acid-locked nucleic acid PCR Clamp
Authors: Hoshi S. , Aoyagi S. , Kono C. , Amano Y. , Yamada Y. , Yamaguchi T. , Ogawa C. , Suzuki N. , Sano Y. , Nukiwa T.
Published: March 2020

20113

Background: EGFR mutations link to the responsiveness to gefitinib, so it is important to get result of EGFR mutations more rapidly and sensitively. Methods: We screened 14 trans-bronchial lung biopsy (TBLB) specimens of non-small cell lung cancer (NSCLC) patients; 10 patients responded to gefitinib and 4 patients did not respond to gefitinib. We examined exons 18 through 21 (the area of the EGFR gene coding for the tyrosine kinase domain) both by the PNA-LNA PCR clamp method and the conventional sequence method. Results: We found five different mutations in 9 patients of 10 who responded to gefitinib by the PNA-LNA PCR clamp method, but only in 7 patients by the conventional sequence method. Detected mutations were the point mutation of exon 18(719S) and exon 21(858R), deletions of exon 19 (E746-A750del (nt2235–2249), E746-A750del (nt2236–2250), L747-E749del A750P). All 4 patients who did not respond to gefitinib were not detected any EGFR mutations either by the PNA-LNA PCR clamp method or by the conventional sequence method. We could get result for 2 days by the PNA-LNA PCR method, but by 7 days by the conventional sequence method routinely. Conclusions: The PNA-LNA PCR clamp method is more sensitive and rapid than the conventional sequence method. The detected EGFR mutations have good relation to gefitinib response.

No significant financial relationships to disclose.
An open-label study to evaluate dose and cycle dependence of the pharmacokinetics (PK) of pegylated liposomal doxorubicin (PLD)
Authors: Gabizon A. , Isacson R. , Rosengarten O. , Tzemach D. , Shmeeda H. , Sapir R.
Published: March 2020

2012

Background: There are no definitive data in humans on the dose dependency and/or cycle dependency of the pharmacokinetics of PLD. The skin toxicity of PLD tends to worsen after several treatment cycles. Therefore, it is important to characterize the PK of PLD after 2 or more cycles. Methods: Fifteen patients with various solid tumors were randomized to two arms of treatment in an open-label study. Arm A received PLD at doses of 60, 30, and 45 mg/m2 in 3 successive cycles every 4 weeks (q4w). Arm B received PLD at doses of 30, 60, and 45 mg/m2 in 3 successive cycles q4w. Twelve patients, 6 on each arm, completed all three cycles and were fully evaluable. Plasma levels of doxorubicin were analyzed by HPLC and fluorimetry following a previously published method (J Chromatogr B, 779:259–69, 2002) with minor modifications. PK analysis was done by non-compartmental method. The following parameters were obtained: Cmax, AUC∞, terminal half-life (T1/2), and Clearance (CL). The paired t test was used for statistical analysis. Results: There was no significant difference in the parameters examined when the dose was increased from 30 to 60 mg/m2. However, when we analyzed the effect of cycle number on the PK, we found a gradual and significant inhibition of CL when patients advance from the 1st through the 3rd cycle of PLD (p=0.0003), with a mean increase of 44% in AUC/mg dose (p=0.0011). Cmax and T1/2 mean values increased by 17% and 18% respectively between the 1st and 3rd cycles, but only the increase in T1/2 was statistically significant (p=0.0127). Conclusion: While the PK of PLD is not dose-dependent within the dose range of 30 to 60mg/m2, there is evidence of a cycle-dependent effect that results in inhibition of CL when patients receive successive cycles of PLD. This effect may account for the delayed skin toxicity of PLD. These results suggest the need for dose adjustments of PLD to minimize the risk of toxicity, such as an initial high loading dose followed by reduced maintenance doses from the 2nd or 3rd cycle onwards.

[Table: see text]

[Table: see text]
Phase I trial of CT-2106 (polyglutamated camptothecin) administered weekly in patients (pts) with advanced solid tumors
Authors: Daud A. , Garrett C. , Simon G. R. , Munster P. , Sullivan D. , Stromatt S. , Allevi C. , Bernareggi B.
Published: March 2020

2015

Background: CT-2106, a camptothecin (CPT) conjugate, is a topoisomerase I inhibitor. Covalent binding through the hydroxyl group of CPT to polyglutamate, a biodegradable polymer of glutamic acid, enhances CPT aqueous solubility and prevents opening of the lactone ring and subsequent CPT binding to albumin. Responses were observed in a Q3 week dosing study in which the maximum tolerated dose (MTD) was 75 mg/m2. Objectives of this study are to determine the MTD and pharmacokinetics (PK) of CT-2106 given weekly to pts with advanced cancer. Methods: Pts received CT-2106 (10 min IV infusion) on days 1, 8, and 15 of each 28 day cycle. Toxicity (NCI CTC v3) and tumor response (RECIST) were assessed. Plasma was analyzed for conjugated and unconjugated CPT by liquid chromatography and tandem mass spectrometry. Results: Enrollment is complete with 21 evualuable pts. Dose limiting toxicities (DLT) are shown in the table . Median age = 60 years (range 36–83), median doses = 6 (1–9), median prior therapies = 3 (1–6). Most pts (12) had melanoma. In a Q3 week study assessments of conjugated and unconjugated CPT demonstrated that CT-2106 provides prolonged exposure to conjugated CPT and slow, progressive release of active CPT from the polymeric backbone. The PK of unconjugated CPT is dependent on the disposition profile of conjugated CPT; unconjugated CPT elimination is formation rate limited. The distribution of conjugated CPT appears to be restricted to extracellular body fluids. Total clearance includes renal excretion documented by substantial urinary excretion levels and CPT cleavage via metabolic pathways. Accumulation of conjugated or unconjugated CPT was not observed with repeated Q3 week dosing. PK in this weekly dosing study will be presented. Conclusions: CT-2106 is generally well tolerated without the toxicities normally associated with CPT, specifically hemorrhagic cystitis and severe diarrhea. The MTD is 25–35 mg/m2 in these heavily pretreated pts.

[Table: see text]

[Table: see text]
Phase I study of NK105, a paclitaxel-incorporating micellar nanoparticle, in patients with advanced cancer
Authors: Kato K. , Hamaguchi T. , Yasui H. , Okusaka T. , Ueno H. , Ikeda M. , Shirao K. , Shimada Y. , Nakahama H. , Muro K. , Matsumura Y.
Published: March 2020

2018

Background: NK105 is a new polymeric micelle carrier system for paclitaxel (PTX). A preclinical study revealed that the plasma AUC and tumor AUC of NK105 were 90-fold higher and 25-fold higher, respectively, than those of free-PTX, i.e., the conventional PTX formulation. NK105 had higher in vivo antitumor activity and lower neurotoxicity than free-PTX. This phase I study was designed to examine the MTD, DLTs, recommended dose (RD) for phase II, and the pharmacokinetics of NK105. Methods: NK105 was administered as a 1-hour intravenous infusion every 3 weeks, without antiallergic premedication. The starting dose was 10 mg PTX equivalent/m2, and the dose was escalated according to the accelerated titration method. Results: To date, 17 patients (pts) have received the following doses: 10 mg/m2 (n=1); 20 mg/m2 (n=1); 40 mg/m2 (n=1); 80 mg/m2 (n=1); 110 mg/m2 (n=3); 150 mg/m2 (n=5); and 180 mg/m2 (n=5). The tumor types treated included pancreatic (n=9), bile duct (n=5), gastric (n=2), and colonic (n=1) cancers. Neutropenia was the most common hematological toxicity. Grade 3 fever developed in 1 pt given 180 mg/m2. No other grade 3 or 4 non-hematological toxicity, including neuropathy, was observed. DLTs occurred in pts given 180 mg/m2 (grade 4 neutropenia lasting for more than 5 days). This dose was designated as the MTD. Allergic reactions developed in only one pt at 180 mg/m2, who was sensitive to other drugs such as antibiotics anti-inflammatory. A partial response was observed in one pt with pancreatic cancer and pts with colonic and gastric cancer had stable disease. The Cmax and AUC of NK105 were dose dependent. The plasma AUC of NK105 at 180 mg/m2 was approximately 30-fold higher than that of the conventional formulation of PTX. Conclusions: Accrual is ongoing at the 150 mg/m2 dose level to determine the RD. DLT was Grade 4 neutropenia. NK105 produces prolonged high levels of PTX in plasma. A 1-hour infusion of NK105 every 3 weeks was feasible, well tolerated, and effective in patients with pancreatic cancer. Even after the long term usage, only grade 1 or 2 neuropathy was observed. NK105 will be evaluated in Phase II studies of patients with advanced pancreatic and gastric cancers.

No significant financial relationships to disclose.
Gemcitabine (G) plamsa and intracellular pharmacokinetics in E6201: Greater metabolite levels using fixed dosing rate (FDR) delivery
Authors: Liebes L. , Levy D. E. , Poplin E. , Mendoza S. , Fry D. , Buckley M. , Zoloratov A. , Benson A. , Hochster H.
Published: March 2020

2024

Background: Gemcitabine (G) is an antimetabolite which requires activation by the rate-limiting enzyme dCK to its active metabolite, dFdCTP. FDR delivery of G at 10 mg/m2/minute may allow greater formation of active metabolite and improved activity. E6201 randomly assigned first line pancreatic cancer patients to three arms: A (G 1000 mg/m2 over 30 minutes), B FDR G (1500 mg/m2 over 150 minutes), and C GemOx (1000 mg/m2 over 100 minutes with oxaliplatin 100 mg/m2). Methods: Investigators from 18 centers contributed 23 sample sets on the first dose of G. Five time points over 4 hrs were sampled (baseline, mid infusion, end infusion, 1–2 post and 4 hrs). Plasma fractionation along with the PBMC purification and subsequent perchloric acid extraction were conducted locally with THU 1 mg (cytidine deaminase inhibitor). Samples were shipped overnight to the central lab where G and its metabolite dFdU were quantified from deproteinized plasma by reverse phase HPLC. dFdCTP was quantified by ion-exchange HPLC in neutralized PBMC extracts after removal of ribonucleotide triphosphates. Data were fit to nonlinear models (WinNonLin, v4.1) and comparisons amongst dosing groups employed the non-parametric, 2-sided Mann-Whitney test. Results: For Arms A, B and C the plasma AUCs were respectively (median ± SD): 4542 ± 2472 (N=9), 8603 ± 2608 (N=8) and 9130 ± 8788 (N=6) ng/ml x hr. The difference between group A and B (p=0.003) and A and C (p=0.05) were statistically different. Intracellular dFdCTP AUCs for groups A, B, and C were: 1862 ± 794 (N=8); 3725 ± 7763 (N=8) and 5024 ± 2113 (N=5) and significantly different for both A vs. B (p=0.05) and borderline significant for A vs C (p=0.065). The latter comparison is limited by a small number of samples. Conclusions: When studied in a multi-centered, randomized cooperative group setting, FDR delivery of G results in higher plasma AUC, and increased intracellular levels of active metabolite dFdCTP. These data support the concept that fixed dose rate delivery of G results in greater intracellular metabolite production at similar and equitoxic doses. Supported by ECOG NYU620HH04–00, NCI CA16087, CA2115, and from Eli Lilly.

No significant financial relationships to disclose.
A phase I dose escalation trial of ispinesib (SB-715992) administered days 1–3 of a 21-day cycle in patients with advanced solid tumors
Authors: Heath E. I. , Alousi A. , Eder J. P. , Valdivieso M. , Vasist L. S. , Appleman L. , Bhargava P. , Colevas A. D. , Lorusso P. M. , Shapiro G.
Published: March 2020

2026

Background: Ispinesib, a novel cytotoxic agent inhibiting the kinesin spindle protein (KSP) has demonstrated significant antitumor activity in multiple murine tumor models. The primary objectives of the study were to assess the safety and tolerability of SB-715992, to determine the dose limiting toxicity (DLT), and the maximum tolerated dose (MTD). Methods: Ispinesib was administered days 1–3 intravenously over 1 hour every 21 days, starting at a dose of 1 mg/m2/day. Traditional 3-patient cohort trial design was utilized with dose levels of 2, 4, 6, 8 mg/m2/day. Results: Twenty-seven patients (24 Caucasians, 3 African-Americans, 16 males, 11 females) with various tumor types were enrolled; colorectal (7), renal (5), bladder (2), lung (2), pharynx (2), pancreas (2), others (7). Grade 3/4 toxicities were noted starting at the 4 mg/m2 dose level with two patients developing grade 4 neutropenia; one for < 5 days, one for > 5 days (with grade 3 leukopenia). At the 6 mg/m2 dose level, grade 3 neutropenia and leukopenia were reported. At the 8 mg/m2 dose level, 3 of 3 patients had grade 4 neutropenia and leukopenia. The 6 mg/m2 dose level was declared the MTD. Toxicities seen in the additional 6 patients included grade 1 fatigue (1/6), grade 1 infusion- related flushing (1/6), grade 3 febrile neutropenia (1/6), and grade 4 neutropenia and leukopenia (1/6). The MTD cohort has been expanded to 10 evaluable patients for confirmation of tolerability and pharmacodynamic endpoints including phosphohistone 3 (PH3), cyclin E, and TUNEL assay on serial tumor biopsies. Preliminary pharmacokinetic data appear linear, but not dose proportional. As predicted, between days 1 and 3, accumulation ranged from 40 to 106%. Exposures appear comparable between cycles 1 and 2. Stable disease in 2 patients with renal cell carcinoma (4 and 5 cycles) and minor response in one patient with bladder cancer were seen. Conclusions: Treatment with ispinesib at the MTD of 6 mg/m2/day x 3 days in patients with advanced solid tumors was well tolerated with consistent dose limiting toxicity of myelosuppression.

No significant financial relationships to disclose.
An open-label phase I dose escalation study of KRN951, a tyrosine kinase inhibitor of vascular endothelial growth factor receptor 2 and 1 in a 4 week on, 2 week off schedule in patients with advanced solid tumors
Authors: Eskens F. A. , Planting A. , Van Doorn L. , Isoe T. , Hayashi K. , Hussain S. , Ekman L. , Burger H. , Verweij J.
Published: March 2020

2034

Background: KRN951 inhibits VEGF induced phosphorylation of VEGF receptors (VEGFR)2 and 1 (IC50 of 0.16 and 0.21 nM) and phosphorylation of c-Kit and Platelet Derived Growth Factor Receptor (PDGFR), (IC50 of 1.63 and 1.72 nM). Methods: The principal objectives of this study were (1) to determine the maximum tolerated dose (MTD) and dose-limiting toxicity (DLT) of KRN951 administered once daily (OD) for 28 days followed by 14 days off treatment, (2) to characterize safety and tolerability, (3) to characterize single and multiple dose pharmacokinetics, (4) to explore inhibitory effects on tumor blood flow, and (5) to look for evidence of antitumor activity. Results: 10 male and 5 female patients, median age 57 yrs (28–72) have been enrolled at dose levels of 1 mg (n=6), 2 mg (n=8), and 1.5 mg (n=1). The total number of courses given is 63 (1–15 per patient) At 2 mg DLT consisting of grade 3 asymptomatic proteinuria, grade 3 ataxia and grade 4 intracranial hemorrhage was seen in three patients. In the next-lower dose level of 1 mg one DLT (grade 3 fatigue) was seen. An intermediate dose of 1.5 mg is currently studied. Hypertension occurred in 14/15 patients but could be medically controlled. Pharmacokinetic analysis revealed dose dependent drug exposure and peak plasma concentrations. Plasma levels of sVEGFR2 decreased following exposure to KRN951. Exploratory analysis by means of Dynamic Contrast Enhanced MRI analysis indicated a decrease in tumorperfusion in selected patients. One confirmed partial response lasting more than 80 weeks in a patient with renal cell carcinoma was seen, and stable disease lasting more than 2 courses of treatment was seen in 6 patients. Conclusion: Once daily KRN951 can be administered safely when given for 28 days followed by 14 days off treatment. The recommended phase II dosing is currently being defined.

No significant financial relationships to disclose.
Paclitaxel and pegylated liposomal doxorubicin in recurrent head/neck cancer: An unexpected administration interval-dependent pharmacokinetic interaction
Authors: Airoldi M. , Cattel L. , Passera R. , Milla P. , Delprino L. , Boselli C. , Buffa C. , Pedani F.
Published: March 2020

2042

Background: Combination of paclitaxel (PTX) with pegylated liposomal doxorubicin (PLD) is an interesting opportunity for recurrent head/neck cancer treatment. Their pharmacokinetic (PK) behavior could be dependent not only on PTX excipient (polyethoxylated castor oil) interference, but also on different iv administration interval between the two drugs. The study endpoint was to evaluate any possible administration interval-dependent PK interaction, when PLD infusion start is delayed from 0 to 24 h after PTX infusion end. Methods: 24 patients affected by recurrent cisplatin pre-treated squamous cellhead/neck cancer were enrolled, receiving PTX 80 mg/m2 q 1w and PLD 12.5 mg/m2 q 2w for 6w/2w rest. Administration interval was 0 h at d1 (PTX-PLD 0) and 24 h at d15 (PTX-PLD 24). Blood sampling was performed at d1–15, PTX and PLD blood levels were analyzed by high performance liquid chromatography techniques, while PK parameters by non-compartmental analysis. Results: PTX PK parameters had large statistically significant differences (median/IQR, PTX-PLD 0 vs. PTX-PLD 24, Mann-Whitney test): Cmax 261/219–531 vs. 407/250–1473 ng/ml p=0.142, AUC 869/688–1331 vs. 3361/969–7853 ng*h/ml p=0.013, Kel 0.39/0.26–0.57 vs. 0.11/0.02–0.26 h⁁−1 p=0.001, Cl 153/89–198 vs. 41/17–138 l/h p=0.013. Similarly, PLD Cmax and AUC were higher in PTX-PLD 24 (Cmax 5.1/3.3–8.1 vs. 6.8/5.3–7.8 mg/l p=0.043, AUC 341/104–1472 vs. 603/106–1006 mg*h/l p=1.000). The overall response rate was 37.5%, including 1 CR (4%); median response duration was 5.5 months (range, 2–16), median overall survival 10 months (range, 2–25+). Conclusions: This exploratory study, having a favourable palliative role in heavily pre-treated patients, showed that PTX PK profile is unexpectedly affected by a different administration interval. In PTX-PLD 0, PTX AUC is fourfold reduced, with a similar increase in Cl, totally due to Kel alteration: therefore, patients could be underexposed to PTX. PLD PK behavior confirmed previous studies results, in which PTX modified PLD disposition, prolonging the duration of its elimination phase and increasing total body exposure to PLD.

No significant financial relationships to disclose.
Pharmacokinetic interaction and safety of enzastaurin and pemetrexed in patients with advanced or metastatic cancer
Authors: Hanauske A. , Weigang Koehler K. , Yilmaz E. , Graefe T. , Kuenen B. , Thornton D. , Lahn M. , Darstein C. , Musib L. , Guiseppe G.
Published: March 2020

2047

Background: Enzastaurin, an orally administered potent PKC inhibitor, is an anti-angiogenic agent that also suppresses PI3K/AKT to inhibit tumor cell proliferation and induce tumor cell death. Preclinical data suggest that the combination of enzastaurin and pemetrexed (Alimta) produced additive or synergistic antitumor activity in tumor specimens. This phase 1b study evaluated the safety, pharmacokinetic (PK) interaction, and antitumor activity of enzastaurin when combined with pemetrexed. Methods: Patients (pts) with advanced or metastatic cancer and an ECOG status of 0–2 received an intravenous dose of 500 mg/m2 pemetrexed on day 1 and an oral dose of 500 mg enzastaurin once daily, (after day 4 in cycle 1) in repeat 21-day cycles for up to 6 cycles. In cycle 1, a loading dose of 1200 mg enzastaurin (400 mg/3×/day) was given on day 4. Pts were also given oral folic acid daily and vitamin B12 every 9 weeks for the duration of pemetrexed therapy, and 5–7 days before cycle 1. Results: Blood samples for PK analysis of enzastaurin (n = 21) were collected on day 21 of cycle 1 and day 1 of cycle 2, and for analysis of pemetrexed (n = 22) on day 1 of cycle 1 and cycle 2. Steady-state concentration [geometric mean (%CV)] of total analytes (enzastaurin and its metabolites) was 1270 nM (126%) when administered alone and 1130 nM (100%) when administered with pemetrexed. Thus, no significant differences in exposures were observed when combined with pemetrexed. Clearance of pemetrexed was the same when dosed either with enzastaurin (2.48 L/h/m2) or as a single agent (2.62 L/h/m2). Safety data from the first 2 cycles are consistent with the known toxicity profile of pemetrexed. No grade 3 or 4 toxicity was reported following the loading dose of 1200 mg enzastaurin. Conclusions: Preliminary data suggest that there is no significant PK interaction between enzastaurin and pemetrexed, and that the combination is well tolerated. The current study is ongoing and subsequent analyses will document tumor responses and safety for longer-term treatment of enzastaurin combined with pemetrexed. This first PK study of enzastaurin and pemetrexed will help determine if enzastaurin has an additive clinical benefit to pemetrexed treatment and support future studies in NSCLC and mesothelioma.

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Bexarotene prevents and reverses gemcitabine resistance in non-small-cell lung cancer cells by affecting gene amplification
Authors: Hermann T. W. , Yen W. , Tooker P. , Ng S. , Negro-Vilar A. , Lamph W. W.
Published: March 2020

2057

Background: Bexarotene (BEX) is a retinoid X receptor selective drug approved for treatment of CTCL and has activity in various cancers. Here we evaluate its effect on acquired drug resistance towards gemcitabine (GEM) in Calu3 NSCLC models. Methods: Parental or GEM-resistant Calu3 NSCLC cells were treated with BEX and GEM as single agents or in combination. Over the treatment course the cells were evaluated for molecular changes (RNA expression and DNA amplification), which were associated with resistance status. Results: Calu3 cells treated with repeated cycles of GEM alone gradually developed resistance to this drug. However, inclusion of BEX prevented the development of resistance and cells remained chemosensitive. Microarray analysis showed a strong increase of rrm1 (ribonucleotide reductase M1) expression in the resistant cells (Calu3-GemR), a gene known to be involved in GEM resistance. In addition, the expression of genes surrounding the chromosomal location of rrm1 was increased suggesting that resistance was due to gene amplification at the 11p15 locus. This was confirmed by analysis of genomic DNA, indicating a 20-fold increase of rrm1 gene copy number. To evaluate BEX’s activity on pre-existing GEM-resistance, Calu3-GemR cells were treated with ten 10-day cycles of vehicle, GEM, BEX, or GEM plus BEX. After 4 treatment cycles the cultures treated with the combination regimen showed a loss in cell numbers, which continued until the end of the 10th cycle. This indicated a re-sensitization of the Calu3-GemR cells to treatment. The cultures treated with the single drug regimens expanded as the vehicle controls. However, analysis of RNA and DNA samples collected over the treatment course revealed that rrm1 gene amplification was gradually reduced even in the cultures treated with BEX alone, reaching parental levels at the end of the study. As expected from these data, the BEX-treated Calu3-GemR cells were again sensitive to GEM. Conclusions: The data indicate that bexarotene can re-sensitize GEM-resistant tumor cells by reversing gene amplification. This suggests that bexarotene may have general clinical utility in cancers where drug-resistance by gene amplification is a major obstacle to successful therapy.

[Table: see text]