bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunotherapy, phase I study of immunotherapy with carcinoembryonic antigen peptide -pulsed, autologous human cultured dendritic cells in patients with advanced non-small cell lung cancer
Authors: Han B. , Huan Z. , Xiaohong F. , Rong L. , Guangli F. , Luoluo J. , Tianqing C. , Wei Z. , Jianzhong S. 
Published: March 2020

12517 Background: To study toxicity tolerance and dose-effect relationship of carcinoembryuonic antigen peptide-pulsed dendritic cells in patients with advanced lung cancer. Methods: cells preparations enriched for autologous DCs were generated from the patients plastic adherent peripheral blood mononuclear cells in media supplemented with granulocyte macrophage colony -stimulating factor and interleukin-4. 37C0, 5%CO2 culture for 7–10. The DCs were loaded with carcinoembryonic antigen(CEA) in day 7, Groups of three to six patients received four weekly or biweekly i.v. infusions of the CAP-1-loaded DC in escalating dose levels of 1 × 106, 1 × 107, and 1 × 108 cells/dose. Patients with lung cancer received iv injections DCs. There were no toxicities directly referable to the treatments. Results: Total 22 patients with lung cancer received DCs immunotherapy. DCs infusion were 2.5×106-9.6×107,means 15.03×106. The early clinical trials suggest that vaccination with CEA vaccines is safe, producing few side-effects, and can lead to CEA-specific immunity. Conclusions: We conclude that it is feasible and safe to generate and administer large numbers of DCs loaded with CEA peptide to patients with lung cancer. No significant financial relationships to disclose.
Src kinase activity in early breast cancer (EBC): A quantitative luminometric and immunohistochemical (IHC) approach
Authors: Young O. , Jones R. J. , Clack G. , Jacobs V. , Marshall A. , Fennell M. , Green T. , Dixon J. M. 
Published: March 2020

13010 Background: Src kinase expression and activity is increased in many solid tumors including breast cancer, and Src kinase has a role in the regulation of cancer cell invasion. Thus, Src kinase is a potential therapeutic target. Targeting cancer cell invasion may be of particular value in early disease, although drug effects are unlikely to be measurable by conventional endpoints, for example tumor regression. This prospective study aimed to establish robust measures of Src activity in invasion-related pathways in tissue derived from EBC and to assess the variability between these measures, in order to establish a methodology for pharmacodynamic assessment of Src inhibition in clinical disease. Methods: Duplicate sets of core biopsies were taken from 30 patients, both at diagnosis and surgery. Biopsies were fixed immediately in either liquid nitrogen (luminometric analysis) or pre-cooled isopentane (IHC analysis). Formalin-fixed, paraffin-embedded tissue (FFPET) was also collected at the time of diagnosis. Levels of phospho-paxillin, phospho-focal adhesion kinase (FAK), total phosphotyrosine and total paxillin were measured in lysed tumor tissue using the Luminex system. Expression of activated Src, phospho-paxillin, phospho-FAK, total paxillin and total FAK was determined in sections of frozen tissue and FFPET, and assessed semi-quantitatively by visual examination. An analysis of variance (ANOVA) model was fitted to the Luminex data ( Table ). Results: All tumors expressed active Src, and there was a high level of expression, but low variability, of paxillin, FAK, phospho-paxillin and phospho-FAK. The subcellular localization of these markers could also be assessed in FFPET sections. Conclusion: The results indicate the feasibility of assessing the effect of a Src inhibitor on invasion-related endpoints in women awaiting surgery for EBC. [Table: see text] [Table: see text]
Systemic telomerase gene therapy delivered via targeted lipidic nanoparticles
Authors: Goldkorn A. , Hann B. , Hayes M. , Drummond D. C. , Kirpotin D. B. , Peth K. , Marks J. D. , Blackburn E. H. , Park J. W. 
Published: March 2020

13025 Background: Most human cancers possess elevated telomerase activity that promotes proliferation. We developed 2 telomerase-targeting gene constructs: telomerase RNA containing a mutated template sequence (MT-hTer), and short interfering RNA (siRNA) targeting endogenous wild type telomerase RNA. When co-expressed in cancer cell lines and xenografts, MT-hTer/siRNA synergizes to cause rapid cell death (Li, S et al. Cancer Research 2004). Here, we tested the in vivo efficacy of MT-hTer/siRNA delivered systemically via antibody-targeted lipidic nanoparticles in a breast cancer metastasis model. Methods: Nude mice received intracardiac injection of luc+MCF7-C18 breast carcinoma cells ectopically overexpressing HER2 and luciferase. After 3 days, the mice were treated with a single 100 μg i.v. injection of either MT-hTer/siRNA-GFP or control (GFP-only) plasmid formulated into lipid-DNA nanoparticles conjugated to an internalizing anti-HER2 single chain antibody (Genosphere, TM) (Hayes, M et al. Gene Therapy 2005). A third group received saline (PBS) as a vehicle control. Cancer progression was monitored non-invasively via biophotonic imaging (Xenogen) of tumor luciferase signal. Results: By week 2 the PBS control group showed luciferase signal elevation that increased exponentially during the study. Similarly, the GFP-only control group developed significant luciferase signal elevation by week 3, with continued rise at all subsequent time points. In contrast, the MT-hTer/siRNA-GFP group experienced a decreased luciferase signal by week 2, which remained consistently low throughout the experimental time course. There were no observed treatment-related toxicities. Conclusions: In pilot in vivo studies, telomerase therapy dramatically inhibited tumor formation in mice after systemic delivery with cancer-targeting lipid-DNA nanoparticles. Expanded preclinical studies are underway. [Table: see text]
Inhibition of histone deacetylase (HDAC) 6 sensitizes human leukemia and breast cancer cells to antagonists of heat shock protein (hsp) 90 and/or bortezomib (BZ)
Authors: Bhalla K. N. , Bali P. , Balasis M. , Fiskus W. , Pranpat M. , Rocha K. , Rao R. M. , Kumaraswamy S. , Atadja P. 
Published: March 2020

13039 Background: HDAC6 is a cytosolic α-tubulin deacetylase, which shuttles misfolded polyubiquitylated proteins into aggresomes. Our studies demonstrated that the hydroxamic acid (HA) analogue pan-HDAC inhibitors (HDIs), e.g., LAQ824 and LBH589, induce α-tubulin and hsp90 acetylation, thereby inhibiting ATP-binding and chaperone function of hsp90. This promotes the polyubiquitylation and degradation of the pro-growth and pro-survival hsp90 client proteins, e.g., Bcr-Abl, mutant FLT-3, c-Raf and AKT in leukemia and Her-2 in breast cancer cells. Methods: Human CML K562 and breast cancer SKBR-3 and BT-474 cells were exposed to LAQ824 (100 nM) or LBH589 (20 to 50 nM) and/or 17-AAG (1 to 5 μM) and/or BZ (5 to 100 nM). K562 and BT-574 cells were also transfected with siRNA to HDAC6. Immunoprecipitation and/or immunoblot analysis were performed to determine the expression or binding of HDAC6 with hsp90, and of hsp90 acetylation and binding with its co-chaperones and client proteins. Immunofluorescent microscopy was utilized to estimate aggresome formation. Cellular cytotoxicity was determined by % inhibition of colony growth. Results: HDAC6 knockdown by its siRNA induced the acetylation of hsp90 and α-tubulin but not of the co-chaperones p23 and hsp40. HDAC6 binds to hsp90. Hsp90 acetylation increased its binding to biotinylated-geldanamycin (GM) and its analogue 17-AAG. Depletion of HDAC6 levels (∼70%) or its activity by treatment with HA-HDI also inhibited the binding of hsp90 to ATP, p23 and hsp90 client proteins, which was augmented by co-treatment with 17-AAG. Knockdown of HDAC6 sensitized the cells to loss of clonogenic survival induced by HA-HDI and/or BZ and/or 17-AAG. Depletion of HDAC6 led to attenuation of Her-2 levels and loss of survival of Her-2 amplified SKBR-3 and BT-474 cells. Co-localization of HDAC6 and polyubiquitylated proteins into aggresomes was enhanced by co-treatment with BZ and/or 17-AAG. This was inhibited by depletion of HDAC6 activity. Conclusions: These findings support the in vivo testing of HDAC6 inhibitors, which through inhibition of hsp90 and aggresome formation sensitizes cancer cells to 17-AAG and/or BZ induced cellular stress due to misfolded and polyubiquitylated proteins. [Table: see text]
Preclinical development of a novel inhibitor of oncogenic B-Raf
Authors: Nolop K. , Jones W. B. , Tsai J. , Cho H. , Bremer R. , Ibrahim P. , Artis D. R. , Hirth P. , Bollag G. 
Published: March 2020

13056 Background: The discovery of oncogenic B-Raf mutations in a majority of patients with metastatic melanoma, and in many tumors from patients with colorectal cancer and other cancers, presents the opportunity to develop oncogene-selective inhibitors with a favorable safety profile. Methods: Guided by co-crystallography, a novel chemical scaffold has been developed into a series of potent inhibitors of oncogenic B-Raf with selectivity versus wild-type B-Raf. From this series, a potent inhibitor has been chosen as a candidate for development. Results: Consistent with the structure-guided approach, this compound shows pronounced selectivity versus a wide array of over 70 other kinases covering all branches of the kinome. This selectivity translates to a wide cellular therapeutic index: inhibition of proliferation of a panel of cell lines bearing the V600E oncogenic B-Raf mutation occurs at IC50s ranging from 40–400 nM while inhibition of cell lines lacking oncogenic B-Raf occurs at IC50s greater than 6500 nM. This cell-based selectivity for the oncogenic B-Raf is greater than the selectivity shown in biochemical assays, supporting that tumor cells bearing the oncogenic B-Raf protein are more dependent on the MAP kinase pathway. Combination experiments with a series of cytotoxic and targeted clinical anti-cancer agents reveal multiple examples of synergistic inhibition of proliferation in vitro, and this synergy generally appears selectively in oncogenic B-Raf-bearing cells. The good oral bioavailability (F > 70%) allows for prolonged exposure in both rodents and non-rodents. Robust efficacy is evident in a murine COLO205 tumor xenograft model, with once-daily oral dosing at 20 mg/kg over 14 days resulting in > 80% inhibition of tumor growth with no effect on body weight. Conclusions: Since this compound is highly selective and targets a B-Raf variant that is absent in all non-transformed cells, it may have a broad therapeutic index, alone or in combination, for the treatment of patients with V600E oncogenic B-Raf tumors. [Table: see text]
A phase 1 study of weekly, divided dose perifosine in patients (pts) with non-small cell lung cancer (NSCLC)
Authors: Henderson I. C. , Spigel D. R. , Nemunaitis J. J. , Richards D. A. , Liebmann J. E. , Purcell W. T. , Birch R. , Gardner L. R. , Burris III H. A. 
Published: March 2020

13063 Background: Perifosine (P) is known to modulate signal transduction pathways, including Akt which is frequently constitutively activated in NSCLC. The half life of P is ∼100 hours and GI toxicities are dose limiting. Studies of a single dose weekly regimen found that the maximum tolerated dose (MTD) was 800 mg. This study was undertaken to see if a higher MTD could be reached using divided doses in NSCLC patients. Methods: Twenty NSCLC patients (pts) were given P, 300 mg, at intervals of 4–6 hours with no more than 4 doses in a 24 hour period. The starting dose was 900 mg and this was escalated by 300 mg/week in subsequent pts to an MTD of 1800 mg. Results: The median age of pts was 62 (range 39–79); 9 were male and median ECOG performance status was 1 (range 0–1). Nineteen pts had received prior chemotherapy (median 2 regimens) and 18 prior radiotherapy. Three pts were entered at each dose and the cohort expanded to 6 pts if 1 or more experienced a grade 3/4 non-hematologic toxicity (DLT) during the first week of therapy. A dose level was toxic if 2 or more pts experienced a DLT. The median time on study was 8 weeks (range 2–28). Seventy-seven percent of pts had no dose reduction (85% of those on 900 mg, 95% on 1200, 72% on 1500, and 78% on 1800 mg). In cohort 1, one pt died within a week without toxicity from P and 1 pt experienced grade 3 nausea, so this cohort was expanded. The other pts in the cohort had no toxicity. In cohort 4 one pt. took 300 mg daily for 6 days and was considered inevaluable. In cohort 3, grade 3/4 toxicity was not observed until after day 28 in 3 pts. Fifteen pts were evaluable for response; 1 had an unconfirmed partial response and 2 had stable disease. Conclusions: This study demonstrated that higher doses of P can be administered on a weekly schedule with divided doses. Weekly doses of 900 and 1200 mg lead to almost no toxicity in most pts. This study has been expanded to a phase 2 trial with direct comparison of weekly and daily doses. The grade 3/4 toxicities for each cohort are given in the table below. [Table: see text] [Table: see text]
A phase I study of axitinib (AG-013736), a potent inhibitor of VEGFRs, in combination with gemcitabine (GEM) in patients (pts) with advanced pancreatic cancer
Authors: Spano J. , Moore M. , Kim S. , Liau K. F. , Hee B. , Bycott P. , Boissaye P. , Francis S. , Peters D. , Rixe O. 
Published: March 2020

13092 Background: Axitinib (AG-013736) is a novel small molecule inhibitor of the receptor tyrosine kinases with picomolar potency against VEGFR 1, 2 & 3 and nanomolar potency against PDGFR-beta and KIT. A phase I study in solid tumors identified 5 mg BID as the therapeutic dose; a phase II study in renal cell cancer demonstrated significant efficacy with a response rate (RR) of 46% (Rini et al, ASCO 2005). This study examined the safety, PK and preliminary efficacy of AG-013736 (AG) in combination with gemcitabine (GEM) as first-line therapy for advanced pancreatic cancer. Methods: A randomized phase II study was preceded by a phase I component. All patients (pts) in the phase I portion received 1000 mg/m2 GEM by 30-minute infusion on days 1, 8, and 15 followed by one week of rest from treatment. AG 5 mg p.o. BID was given beginning Cycle 1, Day 3 (C1D3). Eligible pts had no prior chemotherapy for advanced disease, ECOG 0–2, and no previous treatment with VEGF/VEGFR inhibitors, or anti-angiogenesis treatment. Full PK profiles were collected on C1D1 (GEM alone), C1D14 (steady state, AG alone), and C1D15 (GEM + AG). In the phase II trial, pts are randomized to AG or AG plus GEM beginning C1D1. Results: 8 pts were treated on the phase I portion of this trial. Toxicity: The primary Gr. 3/4 toxicity was hematologic: Gr. 4 anemia and Gr.3 thrombocytopenia in 1 pt and Gr. 3 neutropenia in 1 pt requiring a dose reduction for GEM in Cycle 3. Gr. 2 non-hematologic adverse events include pruritus (1 pt), abdominal pain (2 pts), epigastric pain (1 pt), melena (1 pt), and asthenia (2 pts). Gr. 2 hypertension was observed in 3 pts. Efficacy: Radiological assessment suggests 2 pts with partial response and 4 pts with stable disease: response assessments are ongoing. The median number of cycles is 3 [1,6]. Treatment for 4 pts is still ongoing: Cycle 6 (2 pts) and Cycle 2 (2 pts). Conclusions: This combination is safe and appears to be an effective treatment for advanced pancreatic cancer with significant tumor regression observed in 2 pts. Therapy was well tolerated with manageable toxicity. Additional investigation of AG-013736 in combination with GEM in the phase II setting for advanced pancreatic cancer is warranted. [Table: see text]
Genetic polymorphisms of the adenosine triphosphate-binding cassette transporters (ABCC2, ABCG2) and irinotecan toxicity in cancer patients
Authors: Usui M. , Ando M. , Kitagawa C. , Ando Y. , Sekido Y. , Imaizumi K. , Shimokata K. , Hasegawa Y. 
Published: March 2020

13096 Background: Irinotecan is subject to substantial interindividual variability in pharmacokinetic and the occurrence of unpredictably severe toxicities of leukopenia or diarrhea. These toxicities have been reported to be associated with increased levels of SN-38, the active metabolite of irinotecan. ABCC2 and ABCG2, members of adenosine triphosphate-binding cassette transporters, are involved in mediating the elimination of anionic antitumor drugs, such as SN-38 and SN-38G. Recently, Innocenti et al. reported SN-38G AUC and SN-38G/SN-38 AUC ratios were correlated with ABCC2 3972T>C. The variant ABCG2 421C>A was associated with low ABCG2 expression levels and altered sensitivity to several drugs, including SN-38, in vitro as compared with the reference-type protein. Methods: We assessed whether the variants ABCC2 3972T>C and ABCG2 421C>A would be associated with severe toxicity (leucopenia of grade 4 and/or diarrhea of grade 3 or worse) in 120 Japanese cancer patients in which 27 patients experienced severe toxicity. Results: 74 patients (62%) were homozygotes for the reference allele of ABCC2 3972T>C, 39 heterozygous (33%), and 7 homozygous (6%) for the variant, whereas 62 patients (52%) were homozygous for the reference allele of ABCG2 421C>A, 48 heterozygous (40%) and 10 homozygous (8%) for the variant. Logistic regression analysis did not show any significant associations between the occurrence of severe toxicity and carrying these variants (see Table ). Conclusions: It suggests that genotyping of ABCC2 3972T>C and ABCG2 421C>A would not be useful for predicting severe toxicity caused by irinotecan. [Table: see text] No significant financial relationships to disclose.
Synergistic inhibition of breast cancer cell lines with the combination of a dual inhibitor of EGFR/HER-2/neu and a Bcl-2 inhibitor
Authors: Witters L. , Witkoski A. , Planas-Silva M. , Viallet J. , Berger M. S. , Lipton A. 
Published: March 2020

13100 Background: The epidermal growth factor receptor (EGFR; ErbB1) and HER-2/neu (ErbB2), members of the ErbB family of receptor tyrosine kinases, are overexpressed in a variety of human tumors and overexpression generally correlates with poor prognosis and decreased survival. Use of inhibitors of these receptors as monotherapies, e.g., trastuzumab, Iressa, and erlotinib, has led to advances in treatment, but many patients do not respond or develop resistance. The anti-apoptotic protein, Bcl-2, is also overexpressed in a number of human tumors. Inhibitors of Bcl-2 induce apoptosis and sensitize cancer cells to other therapies. This study assesses the effects of a combination of a reversible inhibitor of both EGFR and HER-2/neu that is similar to lapatinib (GW2974) and a pan inhibitor of the Bcl-2 family (GX15–070: Gemin X Biotechnologies, Inc.) on the growth of human breast cancer cells. Methods: The MCF-7 human breast cancer cell line transfected with a control vector, MCF/neo, and the HER-2/neu transfected MCF-7 cell line, MCF/18, were treated with various concentrations of GW2974 (0.25–10 μM) and/or the GX15–070 pan Bcl-2 inhibitor (50–500 nM). After a 3 day exposure, cell number was determined using the colorimetric MTT tetrazolium dye assay. Percent of control was normalized to corresponding concentrations of the solvent for both agents (DMSO). Results: Treatment with the GW2974 dual inhibitor or the GX15–070 pan Bcl-2 inhibitor resulted in dose-dependent growth inhibition in both the control and HER-2/neu transfected MCF-7 cell lines. The combination of both agents produced synergistic growth inhibition in both cell lines as confirmed by isobologram analysis. Conclusions: This study has demonstrated synergy with the combination of a dual inhibitor of EGFR and HER-2/neu and an inhibitor of Bcl-2 in control and HER-2/neu overexpressing MCF-7 human breast cancer cells. This finding warrants an evaluation of this combination in clinical trials for the treatment of patients with metastatic breast cancer. [Table: see text]
Survivin silencing sensitizes imatinib-resistant CML cells to the cytotoxic effect of hydroxyurea
Authors: Stagno F. , Conte E. , Stella S. , Tirrò E. , Manzella L. , Palio E. , Giustolisi R. , Messina A. , Vigneri P. 
Published: March 2020

13119 Background: Imatinib Mesylate (IM), a semi-specific inhibitor of the BCR-ABL tyrosine kinase, is currently the treatment of choice for Chronic Myeloid Leukemia (CML). However, about one third of CML patients treated with IM develop resistance to the drug because of reactivation of BCR-ABL kinase activity. This phenomenon is usually ascribed to the amplification of the BCR-ABL gene or to the selection of leukemic clones harboring point mutations that abrogate IM binding. To identify novel anti-apoptotic signaling pathways employed by BCR-ABL and devise strategies capable of killing IM-resistant CML cells, we investigated the interplay between BCR-ABL and the Inhibitor of Apoptosis Protein Survivin. Methods: Murine hematopoietic cells (32D) transduced with p210 BCR-ABL and human cell lines either positive (K562, KCL22, KYO1 and LAMA84) or negative (HL60) for the BCR-ABL oncoprotein, were analyzed for Survivin expression by western blot before and after IM treatment. Three different pathways (MAPK, PI3K and JAK2/STA3) potentially involved in BCR-ABL-mediated induction of Survivin were studied using inhibitors specific for each signaling cascade. The effect of Survivin on the proliferation and viability of IM-sensitive and IM-resistant CML cells was investigated after silencing Survivin expression with small interfering RNAs. Results: BCR-ABL tyrosine kinase activity induced an over-expression of Survivin in both human and murine hematopoietic cell lines. This over-expression was both at the transcriptional and the translational level and required the JAK2/STAT3 pathway. Survivin silencing by siRNA increased IM cytotoxicity in IM-sensitive cells but failed to restore IM efficacy in IM-resistant cells. However, Survivin silencing sensitized CML cells to the cytotoxic effect of hydroxyurea and enhanced the efficacy of this compound on three different murine cell lines are insensitive to IM because of point mutations in the BCR-ABL kinase domain (Ba/F3p210Y253F, Ba/F3p210D276G and Ba/F3p210T315I). Conclusions: Reduction of Survivin expression improves the efficacy of IM and increases the sensitivity of IM-resistant CML cells to hydroxyurea. Survivin may represent an attractive therapeutic target for both IM-sensitive and IM-resistant CML patients. No significant financial relationships to disclose.