bluuebunny/crossref_metadata_2025 huggingface dataset:
Breast cancer: Draining lymph node of vaccination site targeted by adjuvant GM-CSF in an autologous vaccine
Authors: Garcia-Giralt E. , Lasalvia-Prisco E. , Cucchi S. , Aghazarian M. , Larrañaga J. , Brent M. , Walton D.
Published: March 2020

13506

Background: We have reported the sensitization against Tumor Associated Antigens (TAA) and the tumor antiprogressive effect of Autologous Thermostable Hemoderivative-Cancer Vaccine (ATH-CV) in breast cancer patients (E. Garcia-Giralt et al ASCO, 2006). Systemic immune response elicited by TAA is started by a locoregional immune response at the microenvironment constituted by the antigens source and the draining lymph node. The antigens source can be a tumor or a vaccination site and respectively, the draining lymph node is the sentinel lymph node (SLN) or the Sentinel Immunized Node (SIN). Tolerogenic or protective locoregional immune response is decisional because it starts a corresponding systemic immune response, tolerogenic or protective. GM-CSF is a conditioner of SLN and SIN, switching the locoregional immune responses from tolerogenic to protective (ML Disis et al Blood, 1996; AJ Cochran et al Nat Rev Immunology, 2006). Therefore, to optimize the ATH-CV antitumoral effect, we have explored GM-CSF as a locoregional adjuvant. Methods: Thirty six M1 breast cancer patients, ER+,HER 2-, hormone and chemotherapy resistant, performance status = 2, and CA 15–3 rising level, were included in this prospective, IRB-approved phase I/II trial. Patients were 3-group (G) randomized (12 each), submitted to different treatments: G1, no additional oncology treatment; G2, ATH-CV; G3, ATH-CV plus GM-CSF at vaccination site 150 μg/day, 5 days. Statistic assessment (Student’s t-test) was performed at the 30-day end point: ATH sensitization by Delayed Type Hypersensibility test (DTH) and Lymphocyte Proliferative Assay (LPA); PSA serum level and immunophenotyping of lymph node cells in biopsies of SIN scintigraphy-localized. Results: Significant differences (p<0.05): DTH test > 5 mm: G1, 0/12; G2, 3/12 and G3, 6/12. LPA > 2.0: G1, 0/12; G2, 4/12 and G3, 7/12. Immunophenotyping: MHC-II+CD83+ (Mature Dendritic Cells):G3>G2>G1. 30 days CA 15–3 increase: G1>G2>G3. No relevant toxicities were evidenced. Conclusions: In advanced breast cancer, ATH-CV sensitization and tumor antiprogressive effect were potentiated by GM-CSF as local adjuvant. Increase of SIN mature dendritic cells is suggested as mechanism of action.

No significant financial relationships to disclose.
Induction of tumor cell death by anti-GD2 monoclonal antibodies (MoAb): Requirement of antibody Fc and a long residence time (slow koff)
Authors: XU H. , Hu J. , Cheung N. V.
Published: March 2020

13507

Background: Anti-GD2 MoAbs have shown clinical activities against neuroblastoma (NB) and other neuroectodermal tumors. Besides complement mediated (CMC) and antibody-dependent cell mediated cytotoxicities (ADCC), they can directly induce cell death. However, MoAb and their genetic constructs differ substantially in death-inducing efficiency. Although multivalent binding is important for these functions, the roles of association on rate (kon), dissociation off rate (koff), valency and Fc have not been carefully defined. Methods: We tested a panel of purified anti-GD2 MoAbs, MoAb fragment, and single chain variable fragment (scFv) fusion protein. The kinetics of in vitro binding was measured by surface plasmon resonance (SPR) using BIACORE 2000. NB cell lines (LAN1, NMB7, BE(2)C, SKNER, SKNJD and SKNLP) were treated with MoAb in vitro, their survivals were studied by the chromogenic assay and IC50 derived using Sigmaplot. Results: The most efficient cell kill against NB was 3F8 (mIgG3) with an average IC50 of 4.7 ug/ml. In contrast, IC50 was >30 ug/ml for 14.G2a (mIgG2a), ME361 (mIgG2a) and S220–51 (mIgG3). It was >100 ug/ml for 3F8 class-switch variant 3G6 (mIgM), 3F8-F(ab’)2(fragment), and 5F11scFv-streptavidin (tetramer). By SPR, the apparent affinity Kd (equilibrium dissociation constant) of 3F8 [1.1x10-8M] was higher than that of 3F8-F(ab’)2 [3.3x10-8M], 14.G2a [9.0x10- 8M], ME361 [4.6x10-7M], and S220–51 [4.6x10-6 M], but lower than that of decameric IgM 3G6 [6.5x10- 9 M] and tetrameric 5F11scFv-streptavidin [2.6x10-9M]. The affinity advantage of 3F8 over other bivalent MoAbs was primarily due to its slower koff [3.0x10-4 sec-1] when compared to other MoAbs [1.1x10- 3-7.3x10-2 sec-1]. Conclusions: Different structural forms of anti-GD2 MoAb have distinct efficiency in inducing NB cell death as well as binding affinity to GD2. Since these properties can affect CMC and ADCC, they may in turn influence the clinical efficacy of immunotherapy. Our findings suggest the importance of an intact IgG3 molecule (the presence of Fc), as well as a long residence time (slow koff).

No significant financial relationships to disclose.
Effect of the immune modulating peptide, SCV-07, in the murine B16 melanoma model
Authors: Tuthill C. W. , Liu Y. , Cai Q. , Wang H. , Kung A.
Published: March 2020

13514

Background: SCV-07 (gamma-D-glutamyl-L-tryptophan) is a synthetic peptide with proven immune stimulating effects and efficacy against various viral and bacterial infections. As several cancers, including melanoma, are known to be sensitive to immune modulation, it was of interest to evaluate the effects of this compound in the murine B16 melanoma model. Methods: Tumors (1x106 B16 melanoma cells) were implanted subcutaneously (sc) into the right axillary area of C57BL/6 mice (Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Beijing, P. R. China). SCV-07 was dissolved in phosphate buffered saline (PBS) and administered once daily via sc administration (at a different site from tumor implantation) at doses of 0.01, 0.10, 1.0, or 5.0 mg/kg for 14 consecutive days. Positive control, dacarbazine (DTIC) was given at a dose of 50 mg/kg once daily for 14 consecutive days via sc injection. Tumor size (by caliper) and body weights were measured once every 3 days throughout the study. The study was terminated on day 17, and the animals euthanized. The tumors were excised, separated, and weighed. Results: SCV-07 treatment led to a significant inhibition of tumor growth compared to vehicle control, with decreases of 16.5% (p=0.030) and 30.2% (p=0.014) seen at the highest doses of 1.0 and 5.0 mg/kg, respectively, on day 15 at the end of treatment. Growth inhibition in the DTIC control was 95.1% (p<0.001). Evaluation of tumor weight also demonstrated a significant inhibition by SCV-07, compared to vehicle, of 23.5% (p=0.008) and 30.8% (p=0.015) at dose levels of 1.0 and 5.0 mg/kg SCV-07. DTIC treated animals had a decrease in tumor weight of 85%. Evaluation of animal body weights demonstrated a significant decrease in those treated with DTIC, suggesting a toxic effect, while no decrease was noted in any SCV-07 treatment groups. Conclusions: Administration of SCV-07 once daily for 14 consecutive days via the sc route is effective against B16 melanoma tumor growth. Tumor growth in animals treated with dose levels of 1 or 5 mg/kg of SCV-07 was significantly reduced compared to the vehicle control group. The overall animal body weights were unchanged after treatment with SCV-07, indicating a lack of toxicity of the test article.

No significant financial relationships to disclose.
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.
SULT1A1 concordance between whole blood and breast cancer tissue
Authors: Grabinski J. L. , Arno E. , Smith L. S.
Published: March 2020

14090

Background: The human sulfotransferase (SULT1A1) enzyme is involved in the conjugation of tamoxifen. Studies have demonstrated the significance of polymorphisms within the SULT1A1 gene in relation to breast cancer survival and tamoxifen metabolism (Nowell et al, J Natl Cancer Inst 2002). DNA for genotyping in previous studies was obtained from a number of different biological specimens including whole blood or paraffin-embedded formalin-fixed tissue. The purpose of this study was to evaluate the genotypic concordance of SULT1A1 polymorphisms in whole blood and breast cancer tissue. Methods: Breast cancer patients receiving tamoxifen and previously genotyped for the SULT1A1 gene were consented for enrollment. Samples representing each of the three possible genotypes (*1/*1, *1/*2,*2/*2) were selected for the collection of archived tissue. DNA was extracted from whole blood using the QIAamp Blood Midi Kit (Qiagen Inc., Valencia, CA, USA) and from formalin-fixed paraffin-embedded breast cancer tissue using the QIAamp DNA Mini Kit (Qiagen Inc., Valencia, CA, USA) as per protocol. Genotyping of the SULT1A1 gene was conducted using a validated Pyrosequencing assay. Results: Tissue samples representing each of the three different SULT1A1 genotypes were compared to the whole blood genotypes. Each of the clinical samples demonstrated 100% concordance between the two different specimens. Conclusion: Our data provides evidence to support the genotyping of clinical samples for the SULT1A1 gene utilizing either whole blood or tumor tissue from breast cancer patients.

No significant financial relationships to disclose.
Functional imaging by DCE-US as a surrogate for response in phase I/II of different targeted therapies by DCE-US: Which quantitative parameter and which timing
Authors: Lassau N. , Koscielny S. , Chami L. , Roche A. , Soria J. , le Cesne A. , Boige V. , Armand J.
Published: March 2020

14093

Background: The early evaluation of targeted treatments is a major challenge in oncology. Functional approaches based on the measure of tumoral vascularization have been developed using different modalities of imaging (CT, MRI, US). We analyzed the response of tumors in three studies using different targeted treatments with dynamic contrast enhanced-ultrasonography (DCE-US). Seven parameters characterizing tumor perfusion were estimated. The objective of the study was to determine which parameter is the most appropriate and when to use it to confirm earlier the efficacy of treatment. Methods: A total of 157 DCE-US were performed in 30 responding patients (PFS>3 months) selected from 3 following studies (multikinase inhibitor targeting angiogenic-receptor with a cytotoxic, thyrosine-kinase inhibitor targeted angiogenic-receptor and C-kit, VEGF monoclonal antibody). Each DCE-US was performed using contrast agent (Sonovue,Bracco) with perfusion and quantification softwares (Toshiba) from raw linear data. Seven quantitative parameters of perfusion were estimated: peak intensity (PI) and area under the curve (AUC), area under the wash-in (AUWI), area under the wash-out (AUWO), time to PI, mean transit time (MTT), wash-in slope. DCE-US were performed before treatment and after during 5 periods (P) : 1–12 days(P1), 13–22 days(P2), 23–43 days(P3), 44–110 days (P4), > 111 days(P5). Results: 1099 parameters have benefited a statistical analysis. Significant modifications (P<0.05) of 4 parameters (PI, AUC, AUWI, AUWO) were observed for the 3 treatments. The earliest significant modifications were observed during the 3rd period for the first 2 studies and during the 4th period for the 3rd study. Conclusions: DCE-US is a sensitive tool to evaluate early tumor response to targeted drugs. Four functional parameters were significantly modified patients responding to treatment: PI, AUC, AUWI, AUWO. Those modifications appear earlier for the multikinase inhibitor targeting angiogenic receptor and the thyrosine-kinase inhibitor targeted angiogenic receptor and C-kit ( (23–43 d) compared to VEGF monoclonal antibody (44 -110 d). DCE- US represent a key add value to early evaluation of targeted therapies.

No significant financial relationships to disclose.
A pilot trial of topotecan administered orally in patients with advanced solid tumors expressing hypoxia inducible factor (HIF)- 1α
Authors: Melillo G. , Gutierrez M. , Holkova B. , Rapisarda A. , Raffeld M. , Horneffer Y. , Chang R. , Murgo A. J. , Doroshow J. H. , Kummar S.
Published: March 2020

14103

Background: HIF-1 is a transcription factor frequently over-expressed in common human cancers that mediates a transcriptional program leading to tumor angiogenesis, survival and metastases. Previous studies have demonstrated that daily administration of topotecan (TPT), a topoisomerase I poison, inhibits HIF-1a expression, angiogenesis and tumor growth in human xenograft models. The primary aim of this study is to investigate whether TPT inhibits HIF-1a expression in human cancers. Methods: TPT was administered orally at 1.6 mg/m2 daily x 5 for 2 weeks, on a 28-day cycle, to adult patients with advanced solid tumors expressing HIF-1a in at least 10% of tumor cells, as assessed by IHC. Tumor biopsies are obtained before and at the end of treatment in cycle 2 and tested for HIF-1a protein expression by IHC and mRNA expression of HIF-1 target genes by real-time PCR. 18FDG-PET, to assess tumor metabolism, and DCE-MRI, to assess blood flow and permeability, were obtained at baseline, at the end of 2 weeks of therapy and then at the end of treatment on cycle 2. CT scans were performed before and after 2 cycles of therapy. Serial blood samples are obtained for PK analysis. Results: Seven patients, median age 54 (range 35–70), have been treated so far, with 6 patients being evaluable for the primary endpoint with paired tumor biopsies for analyses: melanoma (1), colorectal cancer (2), bladder (1), breast (1), ovarian (1). First two patients received the planned dose of 1.6 mg/m2; patient # 2 developed grade 3 neutropenia and grade 4 thrombocytopenia. As the objective was to develop a regimen for chronic dosing that was well tolerated and not directly cytotoxic, the protocol was amended to reduce the dose. Four patients have received a median of 2 cycles at 1.2 mg/m2, which has been well tolerated. A decrease of VEGF mRNA expression by more than 70% was observed in the 2 patients treated at 1.6 mg/m2 after 2 cycles of treatment. However, no consistent decrease of HIF-1a protein levels was detected in the corresponding tumor tissue. Enrollment continues. Conclusion: This is the first study aimed to validate inhibition of the HIF-1 signaling pathway in cancer patients. Funded in part by NCI Contract N01- CO-12400.

No significant financial relationships to disclose.
Association of a novel polymorphism G-133C of thymidylate synthase (TYMS) gene with 5-FU-related severe diarrhea in Japanese patients with colorectal cancer
Authors: Matsumura Y. , Muto M. , Doi T. , Ohtsu A. , Hamaguchi T. , Shimada Y. , Yamada Y. , Shirao K. , Kaniwa N. , Sawada J.
Published: March 2020

14108

Background: 5-FU remains a key drug for the treatment of several cancers, including cancers of the colorectum, stomach, and breast. Increasing evidence suggests that genetic polymorphism results in increased toxicity. This prospective study intended to elucidate the relations between 5-FU-related toxicity and polymorphisms of several genes. Methods: Blood for genotyping was obtained before treatment. Blood samples were drawn at 2-week intervals for 8 weeks to assess hematologic toxicity. Adverse events were assessed according to NCI CTC, v2.0. The genes encoding TYMS and thymidine phosphorylase (ECGF1) were analyzed with leukocyte DNA obtained from patients with colorectal cancer who received 5-FU (500 or 600 mg/m2) and leucovorin (LV) (250 mg/m2) weekly for 6 weeks every 8 weeks. Results: Seventy-eight patients with colorectal cancer were enrolled and genotyped. Toxicity was evaluated in all patients. No genetic polymorphisms detected were significantly associated with 5-FU-related hematological toxicity. Only G- 133C located in the 5’-UTR region of the TYMS gene was significantly associated with 5-FU-related grade 3 diarrhea. Conclusions: The TYMS G-133C SNP may be a critical determinant of severe diarrhea in patients treated with 5-FU/LV. [Table: see text]

No significant financial relationships to disclose.
Effect of enzastaurin on cell signaling, proliferation and apoptosis in breast cancer cell lines
Authors: Spankuch B. , Kurunci-Csacsko E. , Bauknecht T. , Strebhardt K.
Published: March 2020

14121

Background: Enzastaurin, an acyclic bisindolylmaleimide, is a potent selective serine/threonine kinase inhibitor that inhibits PKCβ, targets the PI3K/AKT pathway, and inhibits GSK3β phosphorylation. Enzastaurin induced apoptosis and decreased proliferation of various cancer lines, and decreased VEGF expression and microvessel density in human tumor xenografts. In animal models, enzastaurin had antitumor/antiangiogenic activity in non-small-cell lung, colon, renal cell, hepatocellular, and other cancers. Therefore, we sought to determine enzastaurin’s impact on cellular PKCβ-mediated signaling in breast cancer cells. Secondarily, we sought to determine the induction of the apoptotic cascade by enzastaurin. Methods: Breast cancer cell lines MCF-7, BT-474, MDA-MB-435 and SK-BR-3 were treated with differing enzastaurin concentrations. Western-Blot analyses were performed to examine PKCβ, phospho-GSK3β and caspase 9 expressions. The phenotype and proliferation of enzastaurin-treated cells were also monitored by fluorescence microscopy. Results: Treating all 4 cancer cell lines with ascending enzastaurin doses (0.1–10 μM) led to a significant downregulation of GSK3β phosphorylation (2–17%) compared to control cells. A 48–72 hr incubation with increasing enzastaurin doses also reduced the PKCβ expression significantly (5–50%). Moreover, a dose- dependent reduction of cell proliferation to levels of 15–40% compared to control cells with the highest enzastaurin concentration was detectable. We also saw a marked pro-caspase 9 reduction (0–30%) after enzastaurin compared to control cells. The microscopic inspection of treated cells phenotypically confirmed increasing apoptosis-induced cell death. Conclusions: Enzastaurin has a significant antiproliferative effect in different breast cancer cells. Moreover, enzastaurin suppresses GSK3β phosphorylation, suggesting that it may be a reliable pharmacodynamic marker for enzastaurin activity in breast cancer cells; however, more preclinical analysis is needed. Our study provides evidence for enzastaurin’s potential to directly suppress breast cancer cell proliferation and to induce tumor cell death by apoptotic induction.

No significant financial relationships to disclose.
An open-label phase II clinical trial of PTK787 in patients with progressive neuroendocrine cancer
Authors: Anthony L. B. , McCall J. , Nunez J. , O’Dorisio T. , O’Dorisio S.
Published: March 2020

14127

Background: PTK787 is an orally active angiogenesis inhibitor blocking all known vascular endothelial growth factor receptors (VEGFR) and platelet-derived growth factor receptor (PDGFR)-tyrosine kinases . The majority of metastatic neuroendocrine malignancies express VEGF and PDGF receptors. Since both growth factors have been implicated in tumor-induced angiogenesis, PTK787 offers a novel approach for inhibiting tumor growth. Methods: In a phase II trial, we investigated the safety and antitumor activity of PTK787 in patients who had biopsy-proven metastatic neuroendocrine cancer, such as carcinoid, and had shown evidence of progression on somatostatin analog therapy. Eligible patients had measurable lesions other than bone, a Karnofsky performance status > 60% and normal hematologic, renal and hepatic functions. Patients on octreotide therapy were required to be on a stable dose not exceeding 30 mg monthly of the LAR formulation. Initial dosing of PTK787 was 1,250 mg daily. Results: Eight patients (5 males) were enrolled between 5/20/05 to 9/09/06. Median age was 61 years (range, 48 to 65). There was 1 withdrawal of consent. The median time on study was 6.2 ± 2.1 mos with 1 patient continuing on treatment as of January 2007. Three patients required a 10–28 day discontinuation for rising SGOT/SGPT, alkaline phosphatase and Grade2 proteinuria. Resumption of PTK787 at 1,000 mg daily was well tolerated in 1 patient. One patient developed carcinoid crises with fever, flushing, rising 5-HIAA accompanied by Grade 2 proteinuria. A partial (>50% decrease in 5-HIAA) biochemical response was observed in 25% of the patients. The observed radiographic (CT scans) and scintigraphic (OctreoScan) responses in 6 patients has been stable disease at 3 and 6 mos. Recruitment to this trial continues. Conclusions: PTK787 results in a 25% biochemical partial response rate in neuroendocrine tumor patients with progressive disease following somatostatin analog therapy. Alternative dosing schedules are planned to improve tolerability.

[Table: see text]
Clinical outcome of patients with thymic epithelial tumors
Authors: Palmieri G. , Montella L. , Merola G. , Merola C. , Matano E. , Petillo L. , Evoli A.
Published: March 2020

14148

Advanced chemorefractory thymic epithelial tumors still represent a challenge in clinical oncology. Neoplastic thymoma cells express EGFR. We evaluated the clinical activity of Cetuximab, a chimeric human-mouse monoclonal IgG1 antibody blocking ligand binding to EGFR, in five cases of advanced chemorefractory thymic epithelial tumor. Materials and Methods: Patient Characteristics and response evaluation. Patients’ characteristics and responses to treatment are represented in the table . The patients were assessed for EGFR expression in the primitive tumor, being considered this data as a basis for an anti EGFR treatment. Moreover they were evaluated as concerns clinical status and biochemistry and imaged by a combined PET-CT. Treatment schedule: The patients received intravenous cetuximab (C225, Erbitux, Merck KgaA, Darmstadt, Germany) over 1 hour at 400 mg/m2 loading dose followed by the weekly administration of 250 mg/m2. The first dose of 400 mg/m2 was given during the course of 2 hours. Subsequent weekly treatments were given at a dose of 250 mg/m2 during the course of 1 hour. This preliminary evaluation suggests for the first time that cetuximab may be a useful therapeutic choice in advanced pretreated thymic tumors. Despite the need for prolonged follow-up of the patients and further confirmatory studies, the EGF/EGFR pathway seems to play a pivotal role in thymic tumor. [Table: see text]

No significant financial relationships to disclose.
Circulating tumor cells (CTC) in newly diagnosed breast or colorectal cancers
Authors: Groot M. D. , Croonen M. , Mastboom W. , Vermes I. , Tibbe A. , Tissing H. , Terstappen L.
Published: March 2020

14512

Background: The presence of CTCs is associated with poor prognosis in patients with metastatic carcinomas. The significance of CTCs at the time of diagnosis and after therapy is not known. A prospective clinical trial is conducted to determine if monitoring CTC levels in pts diagnosed with breast and colorectal cancer identify pts at risk for recurrence. CTCs are correlated with pathological stage as follow-up is too short to assess recurrence risk. Methods: Currently 213 pts with newly diagnosed breast and 119 pts with colorectal cancer before surgery with curative intend have been enrolled as well as a control group of 120 patients in whom diagnosis of breast or colorectal cancer was excluded. Thirty ml of blood was screened for the presence of CTCs at baseline before surgery and every following year using the CellSearch™ System. Results: Presence of CTCs in 30mL of blood in breast, colorectal cancer and the control group is provided in the table . In colorectal cancer the average number of CTCs /30 mL at baseline for Dukes A, B and C was 0.2, 0.7 and 1.1 respectively, for stage 0, I, II, III breast cancer 0.3, 0.5, 0.6 and 1.8 respectively and for the control group 0.24 CTCs /30 mL. Conclusions: CTC can be detected in a substantial proportion of pts before and after definitive surgery for breast and colorectal cancer. The number of CTCs before surgery correlated with the stage of the disease. Specificity of the CellSearch™ System is being increased to improve the discrimination of patients with benign and malignant disease. Follow-up must clarify whether the presence CTCs is an identifier for the risk of recurrence.

[Table: see text]

No significant financial relationships to disclose.