bluuebunny/crossref_metadata_2025_split huggingface dataset:
Efficient in vitro expression of the human reverse transcriptase (hTERT) but impaired maturation in dendritic cells of lung cancer patients
Authors: Gahn B. , Brill A. , Bolz I. , Klapper W. , Schub N. , Heiser A. , Gramatzki M. , Kneba M. 
Published: March 2020

13518 An innovative treatment strategy for patients with lung cancer is the targeting of tumor associated antigens with cellular effector cells. The telomerase catalytic subunit (hTERT) is an attractive target for cytotoxic T cells that is highly expressed in both NSCLC and SCLC cells. The aim of the study was the expression of hTERT in dendritic cells (DC) that are the most powerful antigen presenting cells for the induction of cellular immune responses. We used electrotransfection of DC with hTERT mRNA that enables an HLA independent whole antigen approach potentially targeting a wide range of hTERT epitopes. Immature DC were prepared from peripheral blood monocytes in serum-free growth medium, GM-CSF and IL-4. Subsequently the DC were electroporated with mRNA and matured in a cytokine cocktail consisting of IL-1 beta, IL-6, TNF-alpha and PGE2. Verification of hTERT mRNA transfection efficiency was performed by analyzing the induction of telomerase activity with the TRAP assay, the efficiency of GFP electrotransfection was determined by FACS. To optimize mRNA electrotransfection conditions the DCs of 3 healthy individuals were transfected with green fluorescent protein (GFP) mRNA. The percentage of electrotransfected DC was in between 18–89% (mean 46%). Subsequently we electrotransfected DCs in 3 lung cancer patients (1 SCLC, 2 NSCLC). In 1 patient GFP mRNA transfection resulted in 62% GFP positive DC. In the 2 other patients the telomerase activity in the DC 24 hrs after hTERT electrotransfection was equivalent to HL60 cells that biologically express high levels of hTERT. Importantly the DC of one patient did not mature after electrotransfection and incubation in the maturation cocktail. The same was true in another patient, in whom an electrotransfection was not done. In both patients the DC were expressing high levels of HLA-DR and no lineage markers, but CD83 as an indicator of maturation was found in only 1% and 9% respectively. Also the costimulatory molecules CD80 and CD86 were expressed at low levels only. These data show that strong hTERT expression can be achieved in DCs of lung cancer patients using mRNA electrotransfection but demontrate the need for other more potent maturation stimuli. No significant financial relationships to disclose.
Pharmacodynamic and safety study of reolysin in patients with advanced solid tumors
Authors: Gollamudi R. , Desai K. , Chaudhary I. , Ghalib M. H. , Wong B. , Einstein M. , Coffey M. , Gill G. , Mani S. , Goel S. 
Published: March 2020

14004 Background: Reolysin is an intravenous formulation of reovirus serotype 3 - Dearing strain which is a double stranded RNA non-enveloped icosahedral virus capable of inducing cytopathic effects in cancer cells that have an activating mutation in the ras protooncogene. Pre clinical testing has identified cancer cell lines as being susceptible to reovirus infection. Methods: This was an open- label single center phase I safety and dose escalation trial of reolysin administered intravenously over 1 hour on day 0 in a 28 day cycle. Dose escalations were in half log increments. Serum for neutralizing antibody (NA) was drawn at baseline, and days (d) 1, 7, 14, 21, and 28, and serum, stool, saliva, and urine for viral shedding (by RT-PCR) were sampled on d 1,7,14,21, and 28. Response was assessed by imaging studies after 1, 3, 5, and 7 cycles. Results: 18 patients (pt); median age 57 (40–72) years; performance status 0–1; diagnoses - ovarian (6), colorectal (5), and others (7) received 27 cycles of reolysin (median 1, range 1–7) in 6 dose cohorts of 1x108, 3x108, 1x109, 3x109, 1x1010, and 3x1010 tissue culture infective dose (TCID)50. No protocol defined dose limiting toxicities were observed. Drug related grade 2 toxicities included chills (2 pt), and fatigue (3 pt). The only grade 3 event was fever in cycle 7 in a 57 year old woman with progressive breast cancer, treated at a dose of 1x1010 TCID50. She was noted to have 28.5% shrinkage in objective tumor volume. An additional 7 pt had stable disease (SD). No NA was detectable in the serum in any pt at baseline; however all pt developed NA (on d 5 in 5 pt, d 8 in 11 pt, d 14 in 1 pt, and d 28 in 1 pt) during the course of the study. Viral shedding was observed in serum - 5 pt, stool - 3 pt, saliva - 3 pt, and urine - 4 pt. Of interest, 4 of 5 pt with viral shedding had SD as their best response. However, there was no observed relationship between NA formation and SD. Conclusion: Reolysin administered as a one hour infusion on a monthly schedule is safe and well-tolerated even in multiple doses. This preliminary data suggests there is anti-tumor activity of reolysin as a single agent, and warrants further studies either alone or in combination with cytotoxic chemotherapy. [Table: see text]
Modeling targeted inhibition of wild type and mutated c-MET receptor
Authors: Ma P. C. , Jiang S. , Du R. , Dietrich S. , Tang Z. , Kern J. A. 
Published: March 2020

14010 Background: c-MET belongs to the semaphorin superfamily of signaling proteins, containing three protein families (semaphorins, plexins, c-MET and RON) that have central roles in cell signaling. The c-MET receptor tyrosine kinase is involved in regulating cell growth/proliferation, survival, angiogenesis, cell scattering, cell motility and migration. Mutations in c-MET have been identified in various human cancers including lung cancer and papillary renal cell carcinomas. c-MET mutations occur within the extracellular seven-blade β-propeller fold sema domain (E168D, L229F, S325G, N375S), juxtamembrane domain (R988C, T1010I), and kinase domain (M1268T). We hypothesized that these mutations would have differential effects on the kinase inhibition. Methods: We modeled the various c-MET mutations from different functional domains of the receptor using a G418-resistant stable Cos-7 transfection cell system to determine their effect on sensitivity to a selective c-MET kinase inhibitor SU11274. Sensitivity to SU11274 inhibition was assayed by phospho-immunoblotting using phospho- specific antibody against the major tyrosine kinase phosphorylation epitopes pY1234/1235 of the c-MET kinase in vitro. Results: First, we identified that mutations in the sema and juxtamembrane domain were activating as defined by ligand-independent constitutive receptor activation. SU11274 was capable of inhibiting ligand induced signaling through the wild-type c-MET as well as mutant c-MET receptors harboring mutations in the sema, juxtamembrane and tyrosine kinase domain. However, SU11274 inhibition of mutant c-MET was mutation-dependent, with the juxtamembrane domain mutations R988C and T1010I resulting in a receptor form that was less sensitive to SU11274. Mutations in the sema and kinase domain also resulted in varying sensitivity to inhibition by SU11274 inhibition. Conclusions: Mutations in the sema and juxtamembrane domain of c-MET result in receptor activation. The small molecule inhibitor SU11274 is active against wild type and mutated c- MET receptor. Further studies to characterize the signaling effects and the mechanism of sensitivity and resistance of c-MET mutations to specific inhibitors are crucial in the successful development of therapeutic c-MET and mutant c-MET inhibitors. No significant financial relationships to disclose.
A phase I study (CA180021-Segment 2) of dasatinib in patients (pts) with advanced solid tumors
Authors: Johnson F. M. , Chiappori A. , Burris H. , Rosen L. , McCann B. , Luo F. R. , Mayfield S. , Palme H. , Platero J. , Blackwood-Chirchir A. 
Published: March 2020

14042 Background: SPRYCEL™ is a potent, orally active, multi-targeted kinase inhibitor, active against BCR-ABL and SRC family kinases. We report the results of a Phase I dose-escalation study evaluating safety, tolerability, pharmacokinetics, and biomarkers of dasatinib in pts with advanced solid tumors. Methods: Pts with adequate hematologic, renal, cardiac and liver function received oral dasatinib once daily for 7 days per week. Three doses and schedules were examined: 90 mg BID; 140 mg QD; and 180 mg QD. Pharmacokinetics and pharmacodynamic biomarkers were collected on Days 1 and 26 of Cycle 1. Tissue biomarkers were assessed at screening. CT was performed at least every 8 weeks, and FDG-PET at weeks 4, and 8. Results: 26 pts [M=15, F=11] ECOG PS ≤ 2 with epithelial tumors (n=14) or other solid tumors (n=12) have been treated in escalating dose levels. Toxicity was generally mild; most patients came off study for progressive disease. DLTs of pleural effusions were seen in 3/9 subjects on the 180 mg cohort, 2 of whom had pre-existing effusions. Patients with pleural effusion have been excluded from future enrollment. The maximum tolerated dose has not been identified. There have been no objective responses on CT. Six patients have had stable disease with continued study treatment for 2–10 months. Conclusions: Dasatinib can be safely administered at doses of 140 and 180 mg QD. Clinical efficacy, pharmacokinetics, and correlative studies of tumor biopsy material and early FDG-PET results will be reported. No significant financial relationships to disclose.
Insulin-like growth factor I receptor (IGF-IR) inhibition in trastuzumab (T) resistant HER2+ breast cancer (BrCa) cells
Authors: Browne B. C. , Crown J. , Venkatesan N. , Pegram M. , Clynes M. , Duffy M. J. , Slamon D. J. , O’Donovan N. 
Published: March 2020

14049 Background: IGF-IR signaling interferes with the growth inhibitory action of T in BrCa cell lines and may play a role in clinical resistance to T. We studied the effect of IGF-IR inhibition on T resistance. Methods: HER2 and IGF-IR protein levels were determined by ELISA. T growth inhibition of HER2+ BrCa cell lines was measured by cell counting. HER2 and IGF-IR activity were determined by immunoprecipitation with HER2 and IGF-IR antibodies, followed by western blotting with phospho-tyrosine antibody. The effects of IGF- I/IGF-IR inhibition on proliferation and response to T were investigated. Results: No significant correlation was found between HER2, phospho-HER2 or IGF-IR levels and response to T in 12 HER+ BrCa lines. However, two T-conditioned cell lines show resistance to T (BT474 - 2.6 fold; SKBR3 - 1.8 fold) and have higher levels of phospho-IGF-IR compared to parental cell lines. Neither IGFBP3 nor IGF-IR antibody aIR3 significantly inhibited the growth of T-conditioned cell lines or enhanced the anti-proliferative effects of T. IGF-IR siRNA decreased IGF-IR protein levels and inhibited proliferation (29.8 ± 9.3 %) of T-conditioned SKBR3 cells. IGF-IR siRNA also enhanced T growth inhibition of T-conditioned SKBR3 cells (47.7 ± 13.7 % compared to 17.7 ± 14.8 % for T alone). The small molecule IGF-IR tyrosine kinase inhibitor (TKI), NVP-AEW541, inhibited growth of parental and T-conditioned SKBR3 cells and enhanced response to T (Table). Conclusions: Increased activation of IGF-IR may play a role in resistance to T. Inhibition of IGF-I ligand binding does not appear to alter IGF-IR mediated T-resistance. However, reducing IGF-IR protein levels or blocking IGF-IR TK activity improves response to T in resistant cells. Thus, the combination of IGF-IR TKIs, such as NVP-AEW541, with T may be beneficial for T-resistant HER2+ BrCa. [Table: see text] [Table: see text]
Optimal safety profile and anticancer activity of NGRhTNF coupled with doxorubicin (NGR003 modified phase I combo trial)
Authors: Rizzardi G. , Gregorc V. , Vitali G. , Gallo Stampino C. , van Herpen C. , Santoro A. , Sobrero A. , Punt K. , Caligaris-Capio F. , Bordignon C. 
Published: March 2020

14056 Background: NGRhTNF is a vascular targeting agent (VTA) exploiting a tumour homing peptide (CNGRCG) selectively binding angiogenic vessels in solid tumors, where NGRhTNF specific binding relies on dynamic interactions with TNF-receptors and aminopeptidase N (CD13). NGRhTNF combines activity on tumour vascular permeability and direct anticancer activity. At low dose, NGRhTNF increases tumour vascular permeability. Consistently, mouse preclinical data indicate significant synergy between low dose NGRhTNF and cytotoxic agents. Thus, we tested NGRhTNF in combination with doxorubicin. Methods: 4 doses of NGRhTNF (0.2, 0.4, 0.8 and 1.6 μg/m2) in combination with doxorubicin (75 mg/m2) have been administered every 3 weeks in 15 patients.Main end- points included safety, anticancer activity and pharmacokinetic. Measurement of circulating tumour (CTC) and endothelial cells, sTNF receptors, along with plasma chemokine profile are being performed. Results: All patients were enrolled (5F/10M). Toxicity was limited to constitutional symptoms, most frequently chills. After a median follow-up of 12 weeks (range 3–28), 5 episodes of chills (1 grade I and 4 grade II) were reported in 4/15 patients.One episode of grade I hypertension was reported. Of note, the toxicity profile commonly-associated to doxorubicin has not been exacerbated. Stabilization of disease occurred in 10/15 patients lasting 13 weeks (range 6–28 with 5 patients still ongoing at December 2006). In addition, partial response was achieved in 2 patients, lasting 23 and 14 (still ongoing) weeks, respectively. It is worth noting that anticancer activity was observed in 12/15 patients of whom 9 had previously received anthracycline-containing regimens and 5 were anthracycline-resistant suggesting a role of NGRhTNF in inducing tumour sensitivity to doxorubicin. Finally, changes in CTC levels over time consistently matched the clinical outcome. Conclusions: Coupling NGRhTNF with doxorubicin induced anticancer activity along with an optimal safety profile in pretreated patients providing support for the phase II combination studies addressing diseases suitable for anthracycline treatment such as sarcoma and ovarian cancer, and planned to start in early 2007. No significant financial relationships to disclose.
Preclinical modeling of a phase 0 clinical trial protocol
Authors: Kinders R. J. , Hollingshead M. , Parchment R. E. , Khin S. , Kaur G. , Phillips L. , Tomaszewski J. , Doroshow J. 
Published: March 2020

14058 Background: The Exploratory IND program initiated at NCI (“Phase 0”) is designed to evaluate the pharmacodynamic effects (PD) of candidate drugs at the molecular level in the clinic. Trials employ patient biopsies and surrogate tissues (e.g. PBMCs) to determine the quantitative effect of the agent on its putative target, after a minimum number of doses. The approach requires repeated biopsy of the tumor, an understanding of the time-effect window, and some knowledge of the dose level likely to cause a measurable drug effect. Methods: Prior to initiating a Phase 0 study of the PARP (PolyAdenosyl-Ribose Polymerase) inhibitor, ABT-888, we developed a pre-clinical model to mirror the clinical protocol. Colo 829 and A375 xenografts in athymic nude [nu/nu (NCr)] mice were examined for time and dose effect on PARP using a validated, quantitative PAR assay. Extracts of entire xenografts, quartered xenografts and 18 gauge needle biopsies were examined for variability of baseline and post-treatment PAR levels. Results were cross-checked with Western analysis for polyADP-Ribose (PAR)-labeled proteins in treated mice. Pharmacokinetics (PK) were modeled using plasma drug levels. Additional studies examined the influence of previous biopsy, contralateral biopsy, vehicle treatment, and general anesthesia on PAR in xenografts. Results: A single dose of ABT-888 produced a significant decrease in intracellular PAR levels that could be measured 2 to 6 hours post-dose. PAR levels and drug effect on PAR levels were not influenced by repeated needle biopsies. Variation across xenografts was random for single and bilateral xenograft animal models in the ABT-888 treated, vehicle- and topotecan-treated control groups. Animal handling and socialization appeared to elevate baseline PAR levels, which could confound analysis of study results. Conclusions: Pre-clinical modeling of a specific Phase 0 clinical protocol for drug effects and biological variability provided valuable insights into the development, refinement, and analysis of the currently-active NCI Phase 0 clinical trial of ABT-888. Animal studies were conducted in an AAALAC approved facility under an approved IACUC protocol. Funded by NCI Contract N01-CO-12400. No significant financial relationships to disclose.
Combined EGFR targeted therapy in a novel in vivo pancreas cancer model
Authors: Jimeno A. , Chan A. , Rubio-Viqueira B. , Zhang X. , Cusatis G. , Wheelhouse J. , Varella-Garcia M. , Hirsch F. R. , Kulesza P. , Hidalgo M. 
Published: March 2020

14063 Background: EGFR inhibitors have a definite but limited activity in pancreatic cancer (PaCa). We have reported enhanced activity of dual EGFR therapy with a small molecule inhibitor (erlotinib) plus a monoclonal antibody (cetuximab) in head and neck cancer models. Human xenografted tumors recapitulate better the pathobiology of cancer than existing cell lines. Here we explored a dual EGFR targeting approach using a direct PaCa xenograft model, and sought after markers predicting efficacy. Methods: PaCa specimens obtained at the time surgery were implanted in nude mice and expanded to develop cohorts of tumor bearing mice suitable for drug evaluation. Ten cases were expanded, and within each case 4 groups of 6–8 mice each were treated with vehicle, erlotinib, cetuximab, and the combination of both for 28 days. Gene mutation analysis, gene amplification by fluorescence-in-situ hybridization, and immunohistochemistry (IHC) were done with untreated samples. Results: Two cases were sensitive to both single agents, but the combination did not induce higher efficacy in those. Two additional cases that were resistant to both single agents became sensitive with the combination. No egfr mutations were detected. Three and four cases carried low and high egfr polysomy (defined as [[Unsupported Character - ]] 4 copies in 10–40% and [[Unsupported Character - ]] 40% of the cells, respectively). No correlation was found between egfr copy number and efficacy. By IHC sensitive cases had a lower Ki67 proliferation index, and higher EGFR and nuclear pMAPK staining than resistant cases. The degree of Ki67 decrease after therapy correlated with efficacy. In cases resistant to the single agents but sensitive to the combination nuclear pMAPK only decreased with the dual targeting. Cases with high egfr polysomy were more labile to pharmacodynamic effects after treatment (such as EGFR or pMAPK decreases). Conclusions: EGFR inhibitors showed modest single agent antitumor effect that was enhanced with dual EGFR therapy in PaCa. No genomic markers predicted efficacy, although high egfr polysomy was associated with deeper pharmacodynamic inhibition, conceivably suggesting a phenomenon of pathway addiction. Higher pathway activation by IHC was linked with higher activity. No significant financial relationships to disclose.
Epidermal growth factor receptor (EGFR) inhibition in triple-negative breast cancer (BrCa)
Authors: Corkery B. , O’Donovan N. , Clynes M. , Crown J. 
Published: March 2020

14071 Background: Triple-negative BrCa lacks expression of hormone receptors and HER-2 but does express EGFR. It is associated with early relapse and poor survival. There is no targeted therapy for triple-negative BrCa. We are studying the potential role of EGFR inhibition. Methods: EGFR expression was examined in triple-negative BrCa cell lines, (BT20, HCC1937, and MDA-MB-231) by western blot. IC50 assays were determined for three EGFR inhibitors, gefitinib (G) and erlotinib (T), which are small-molecule tyrosine kinase inhibitors, and cetuximab (E) which is a monoclonal antibody against EGFR, and chemotherapy (CRx) drugs docetaxel (D) and carboplatin (P)), using the acid phosphatase assay. The controls were HER2+ BrCa cell lines, BT474 and SKBR3 which express low levels of EGFR. Results: The three triple-negative cell lines over-express EGFR. IC50 values for G and T were significantly higher in the triple-negative than in the HER2+ cell lines. E did not cause significant inhibition in any cell line (max inhibition 20% at 100 μg/ml E). IC50 values for G were lower than for T in the triple-negative cell lines (IC50s for HCC1937: G - 8.4 ± 1.5 μM; T - 26.2 ± 9.3 μM). Combined EGFR inhibition with CRx was tested in HCC1937 cells. G combined with P or with D for 5 days showed an additive effect on inhibition of proliferation ( Table 1 ). Alternate scheduling of the drugs did not significantly influence response. Conclusions: Our results suggest that triple-negative BrCa cells over-express EGFR but are not as sensitive to EGFR inhibition as HER2+ BrCa cells. However, EGFR inhibition may enhance response to CRx in triple-negative BrCa. [Table: see text] No significant financial relationships to disclose.
Over-expression of the A3 adenosine receptor in tumor and PBMC of hepatocellular carcinoma patients: A specific therapeutic target
Authors: Fishman P. , Stemmer S. , Castel D. , Ochaion A. , Cohn S. , Barer F. , Zabutti A. , Bar Yehuda S. 
Published: March 2020

14085 Background: The Gi protein associated A3 adenosine receptor (A3AR) is highly expressed in colon carcinoma tissue and is reflected in peripheral blood mononuclear cells (PBMC) of the patients. Drug candidates with high affinity to the receptor possess anti-cancer effect when administered orally to colon carcinoma bearing animals. The receptor was suggested as a potent therapeutic target and predictive biological marker. Since there is no adequate therapy for Hepatocellular Carcinoma (HCC) we looked at the presence of A3AR in tumor and PBMC of HCC patients and tested the efficacy of the A3AR agonist, CF102 in an experimental rat HCC model. Methods: mRNA and protein A3AR expression level were examined in paraffin embedded slides and PBMC from HCC patients utilizing RT-PCR and Western blot (WB) analysis, respectively. Rat HCC model was established by inoculating N1S1 cells to the animals’ liver. Oral treatment with CF102 was initiated 5 days after tumor inoculation. Signaling proteins in tumor lesions and PBMC from the animals was tested by WB analysis. Results: A3AR over-expression was found in tumor and PBMC of HCC patients. Receptor over-expression was attributed to high expression of NF-κB, known to act as a transcription factor of A3AR. In the HCC bearing rats, A3AR was over-expressed in the tumor tissue and PBMC. Upon treatment with CF102 A3AR expression was down-regulated followed by inhibition of tumor size (60%±18.4%, p<0.05). Mechanistically, CF102 de-regulated the NF-κB and Wnt signaling pathways. Conclusions: A3AR which is up- regulated in HCC tumors may be suggested as a new target to treat this malignancy and as a biological marker to follow up tumor response to the drug. No significant financial relationships to disclose.