bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase I safety and tolerability of once daily oral afatinib (A) in combination with gemcitabine (G) in patients with advanced solid tumors.
Authors: Zanetta Sylvie , Bennouna Jaafar , Isambert Nicolas , De-Montserrat Helene , Squiban Patrick J. , Delord Jean-Pierre
Published: March 2020

e13009

Background: A is an orally bioavailable, irreversible, ErbB Family Blocker. This open label, Phase I, dose escalation trial investigated the safety, tolerability and pharmacokinetics of A in two parallel dose cohort expansion parts, in combination with either G (Part A) or docetaxel (Part B) in patients with relapsed or refractory solid tumors. Preliminary results from Part A are presented here. Methods: Eligible patients (confirmed diagnosis of advanced solid tumors, ECOG PS 0–1) received once-daily, oral dosing of A in combination with G, given intravenously at Day 1 and at Day 8 of every 3 week cycle. Dosing of A started on Day 2 of Cycle 1. The primary objective was to establish the maximum tolerated dose (MTD) based on the occurrence of dose limiting toxicities (DLTs) observed in Cycle 1. Dose escalation was performed with cohorts of 3–6 patients using a 3+3 design. Initial starting dose level was A 30 mg/day and G 1000mg /m², escalating up to A 50 mg/day and G 1250 mg/m², until the MTD was reached, and followed by a PK expansion cohort of 12 patients at the MTD level. Incidence and severity of AEs were recorded. Results: To date, 19 patients have been treated on study with the following baseline characteristics: mean age (53.7 years), women (63.2%) and number of prior chemotherapies (≤2: 26%, >2: 74%). Twelve patients received 2–4 cycles of treatment and five patients received 4 or more cycles. AEs observed in most patients were diarrhea (89.5%) and rash (63.2%). In Cycle 1, DLTs were experienced by one patient out of six receiving A 30 mg and G 1250 mg/m². MTD was exceeded at a dose level of at least A 40 mg/day and G 1250 mg/m². An intermediate dose level of A 40 mg/day and G 1000 mg/m² is currently under evaluation. Conclusions: In patients with relapsed or refractory advanced solid tumors, the combination of A with G is well tolerated, with manageable AEs. Dose finding is ongoing and MTD, safety profile and preliminary evidence of activity are anticipated to be reported at time of presentation.
Personalizing cancer treatment of combined targeting of PI3K/AKT and MAPK pathways in advanced cancer patients harboring simultaneous oncogenic alterations in both signaling pathways.
Authors: Shimizu Toshio , Tolcher Anthony W. , Papadopoulos Kyriakos P. , Beeram Muralidhar , Rasco Drew Warren , Smith Lon S. , Drengler Ronald L. , Gunn Shelly , Smetzer Leslie , Mays Theresa A. , Kaiser Brianne , Wick Michael Jude , Patnaik Amita
Published: March 2020

e13025

Background: Frequent coactivation of both PI3K/AKT and MAPK pathways has been seen in a number of different tumor types. Preclinical studies also provide a clear rationale for the co-inhibition of these, "semi-parallel" pathways. Methods: We reviewed the records of 1,672 consecutive advanced cancer patients (pts) in our Phase I Clinical Trials Program and investigated the clinical impact of simultaneous blockade of both PI3K/AKT and MAPK pathways in patients with oncogenic alterations in both signaling pathways from 317 pts who received PI3K pathway inhibitor and/or MAPK pathway inhibitor. Results: 163 of 317 pts (51.4%) were tested for comprehensive tumor genomic analysis using DNA array-based CGH/PCR-based DNA sequencing. PI3K pathway genetic alterations (PIK3CA mutation, n=3; PTEN deletion, n=18; AKT amplification, n=10) were detected in 29 of the 163 (17.8%) pts. RAS/RAF pathway genetic alterations (KRAS mutation, n=33; HRAS mutation, n=4; BRAF mutation, n=6) were detected in 43 of the 163 (26.4%) pts. Simultaneous oncogenic alterations in both PI3K/AKT and MAPK pathways were detected in 12 pts (colorectal cancer, n=7; pancreatic cancer, n=2; melanoma, n=2; non-seminomatous germ cell tumor, n=1). Six of 8 (75%) pts treated with personalized treatment based on dual pathways inhibition had tumor regression with prolonged duration of time to treatment failure compared to that of pts treated with single pathway inhibition. A pt with pancreatic cancer harboring simultaneous KRAS mutation and AKT2 amplification treated with combination MEK1/2 inhibitor and AKT inhibitor showed central cavitations of numerous lung metastatic lesions along with a decrease in CA19-9, which may reflect treatment effect. Conclusions: These data could serve as a platform for therapeutic decision making. The strategy of targeting parallel pathways may be especially important in pts with coexisting PI3K/AKT and MAPK pathway genetic alterations. The data also suggest the need to further investigate the role of molecular profiling and matching pts with targeted drugs for personalized treatment.
CGTG-102 (Ad5/3-D24-GMCSF), a novel oncolytic adenovirus, in patients with refractory solid tumors: Experience from an advanced therapy access program.
Authors: Von Euler Mikael , Kanerva Anna , Nokisalmi Petri , Koski Anniina , Diaconu Iulia , Cerullo Vincenzo , Joensuu Timo K. , Oksanen Minna , Pesonen Sari , Kangasniemi Lotta , Ranki Tuuli , Heiskanen Raita , Hemminki Akseli
Published: March 2020

e13035

Background: Following preclinical testing CGTG-102, a 5/3 chimeric oncolytic adenovirus armed with human GMCSF, has been used to treat 115 refractory cancer patients. Methods: Eligibility criteria included refractory advanced solid tumors, no major organ deficiencies and written informed consent. Patients were treated with either a single treatment or serial treatments with one or more viruses. Intra tumoral administration was performed under ultrasound guidance. The initial dose, 8 x 1010 Viral Particles (VP), was based on published safety results and preclinical testing and escalated in later patients. A routinely tolerated dose of 3 x 1011 VP was deemed optimal and is the target dose for clinical development. To reduce regulatory T-cells, low-dose cyclophosphamide 50 mg/day was given. Adverse Reactions (AR) were scored according to CTCAE 3.0. Imaging was done by CT before and ~2 months after treatment. Response was scored according to RECIST 1.1, including injected and non-injected lesions. Decrease not fulfilling PR was scored as minor response (MR). Results: The most common ARs were pain (82%), fever (81%), fatigue (79%), nausea (54%) and hemoglobin decrease (48%). Pain is mostly tumor pain or pain in the injected tumor, which may be causally related to the MOA of the therapy. Most ARs were G1 or G2; 6 were G4: 2 Hb decrease, 2 pulmonary embolism and single reports of thrombocytopenia and pericardial effusion, most probably due to the underlying disease.Imaging was performed when clinically useful. 65/115 are evaluable by imaging: 3% PR, 11% MR, 40% SD and 46% PD. Best results were obtained in Breast Cancer, Melanoma, Soft Tissue Sarcoma, Mesothelioma and Ovarian Cancer. Median survival in this heavily pre-treated refractory population is 164d, 95% CI 122d – 206d. Mean survival is 281d reflecting that approx. 30% survive more than 300d and 15% up to 600d. A wide range of samples are being analyzed to further characterize the viral and immunological aspects of the therapy. Conclusions: CGTG-102 is a novel oncolytic adenovirus with good safety profile and encouraging signs of efficacy. Formal clinical studies are underway in several tumor types in both US and EU.
Phase I clinical trial of multi-antigen peptide vaccines therapy using cancer-testis antigens for patients with advanced or recurrent breast cancer.
Authors: Fujita Tomoyuki , Yoshida Atsushi , Nishimura Hajime , Koshikawa Kayoko , Nagura Naoya , Yoshida Koji , Miyoshi Yasuo , Yamauchi Hideko , Nakamura Seigo , Nakamura Yusuke , Fujimori Minoru
Published: March 2020

e13037

Background: Cancer-testis (CT) antigens are proteins that are normally expressed only in the human germline, but are also present in a significant subset of malignant tumors. The practical importance of these proteins is that, due to their restricted expression pattern, they are frequently recognized by the immune system of cancer patients. Moreover, this antigenicity has raised the possibility of their use as vaccines to stimulate immune responses in order to combat tumor growth. As a result, many aspects of CT antigens have been examined in studies performed worldwide. Here we report the results of a phase I clinical trial combining five kinds of CT peptide (CDCA1,URLC10,KIF20A,DEPDC1,MPHOSPH1) vaccination. Methods: The subjects of the trial were HLA-A24-positive patients with metastatic and advanced breast cancer. The mixture of peptides was subcutaneously injected weekly with dose-escalation (doses of 0.5, 1, and 2 mg of each peptide/body, three patients/cohort). Safety, efficacy and immunological parameters were assessed. Results: No adverse events were observed in any patients. Of the 9 patients who completed at least one cycle of the treatment, 4 (44%) developed immunological reactions at the injection site. Specific cytotoxic T lymphocytes (CTLs) reacting to any peptide were induced in 7 (78%) of the 9 patients. Of the 7 patients who completed at least two cycles of the treatment, tumor regression compatible with stable disease (SD) was noted in 4 patients. The remaining patients had progressive disease (PD). The vaccine therapy was tolerable and safe at all doses. Conclusions: Combination therapy with CT antigens has therapeutic potential for patients with breast cancer. Peptide-specific CTLs against peptides in the vaccine were induced at a high rate, even in heavily pretreated patients. From an immunological perspective, the optimal dose for further clinical trials is ≥1 mg/body.
Role of chronic macrophage activation in patients with primary breast cancer.
Authors: Thein Mya Sanda , Jain Alka , Ingaramo Maria Clara , Kohli Antia , Fedarko Neal
Published: March 2020

e13042

Background: Breast cancer progression has been associated with altered immune function and macrophage activation. Neopterin (Neo), a GTP metabolite, is secreted by interferon-γ activated macrophages. Chitotriosidase (ChT), a member of the glycosyl hydrolase family, is secreted by chronically activated macrophages. Osteopontin (OPN), a phosphorylated glycoprotein, produced by tumor and immune cells, is a modulator of macrophage activity and infiltration. OPN and neopterin have been studied separately as markers of solid malignancy including breast cancer. Methods: The purpose of our study was to test for correlations between breast cancer presence and stage with macrophage produced markers of acute (Neo), and chronic (ChT) macrophage activation as well as tumor/immune cell produced marker of macrophage infiltration (OPN). The study included 66 patients with primary breast cancer (Group I: 14 with metastatic disease) and 111 participants in a control group (Group II). Serum levels of OPN and Neo were measured by ELISA, while ChT levels in serum were measured by enzyme activity assays. Comparisons between groups were analyzed by Mann Whitney U test. Results: ChT and OPN levels were significantly higher in Group I compared to Group II (Table I). ChT was elevated at early stage disease while OPN increased with more advanced disease. Although Neo was not increased in patients with breast cancer, a subgroup analysis within group I demonstrated that it was higher in patients with metastatic disease (6.8 vs 9.3 p <0.01). Conclusions: High levels of ChT in breast cancer patients may indicate that chronic macrophage activation plays a significant role in early breast cancer progression. OPN and Neo, elevated in late stage and metastatic disease, respectively may be useful biomarkers of tumor aggressiveness and/or metastatic progression. Further studies should be done to validate these biomarkers and understand the role of acute and chronic macrophage activation in cancer progression. [Table: see text]
Open label phase I/II study of Wilms' tumor gene 1 (WT1) mRNA-transfected autologous dendritic cell vaccination in patients with solid tumors.
Authors: Willemen Yannick , Huizing Manon Thirza , Smits Evelien , Anguille Sébastien , Nijs Griet , Stein Barbara , Van Tendeloo Viggo , Peeters Marc , Berneman Zwi N.
Published: March 2020

e13051

Background: Active specific immunotherapy, aimed at stimulating tumor-specific immunity, is widely under investigation to determine its place in cancer treatment. Methods: We are conducting an open label phase I/II clinical trial to evaluate the feasibility, toxicity and immunogenicity of intradermal vaccination with keyhole limpet hemocyanin (KLH)-exposed WT1 mRNA-transfected autologous monocyte-derived dendritic cells (DC). Recruitment has finished, but vaccination and follow-up continue. Vaccination consists of biweekly intradermal injection of 10 million DC in the medial region of the arm close to the axilla. Clinical evaluation with PET, CAT or MRI is done before start of the vaccination and repeated every 8 weeks. After 4 vaccinations patients undergo a delayed-type hypersensitivity (DTH) skin test to evaluate immunological responses. Results: 18 patients (8 breast cancer and 3 astrocytoma patients, 1 melanoma, 1 mesothelioma, 1 ovarian, 1 colon, 1 esophageal, 1 renal cell and 1 peritoneal cancer patient) were included in this trial from May, 2010 until January, 2012 to whom we administered a total of 175 vaccines. The median number of vaccines produced per patient was 15 (range 5-34). Median follow-up from start of the vaccination is 26 weeks (range 2-83). All evaluable patients exhibited local symptoms such as itching and/or redness at the sites of injection after the second vaccination and onward (n=17) and displayed positive DTH skin reactions (induration ≥2 mm) to the vaccine and its components (n=15). Systemic side effects were limited to 2 patients who each experienced a single episode of flu-like symptoms within 24 hours after vaccination. Thus far, 3 patients died of disease progression, while 7 others had progressive disease and 6 had stable disease at the most recent evaluation compared to before vaccination according to RECIST 1.1. Of the latter group 4 patients received another form of therapy at some point during vaccination. Conclusions: Based on these results our WT1 mRNA-transfected DC vaccination appears to be feasible and safe for patients with solid tumors. Furthermore, our vaccine is capable of inducing immune responses in these patients.
Tailored endpoints: A proposal for design of future clinical trials in metastatic breast cancer (MBC).
Authors: Fontanella Caterina , Bonotto Marta , Driol Pamela , Valent Francesca , Gerratana Lorenzo , Russo Stefania , Andreetta Claudia , Minisini Alessandro , Mansutti Mauro , Sottile Roberta , Fasola Gianpiero , Puglisi Fabio
Published: March 2020

e13058

Background: Widespread access to active agents against MBC has produced significant improvements in survival, even though few phase III trials are powered to detect overall survival (OS) effects. However, regulatory agencies hold OS as gold standard for approval of new drugs. OS can be portioned into the sum of progression-free survival (PFS) and survival post-progression (SPP). Recent data suggest that OS is a reasonable primary endpoint only when median SPP is short. On the other hand, patients enrolled in first-line trials could have a long life-expectancy. Aim of this study was to evaluate PFS, OS and SPP in a consecutive series of patients with MBC in order to obtain information as a basis for design of future clinical trials. Methods: Four hundred patients with MBC diagnosed from January 2004 to February 2011 at University Hospital of Udine, Italy, were included in the study. We examined the role of potential of disease and patients’ characteristics in influencing the different measures of outcome. Results: Median OS was 33.71 months (mo), in line with the best literature. Median PFS at first-line (PFS1) was 9.33 mo, with linear decrease at second (5.06), third (3.58), and fourth (3.19) line, respectively. Median SPP after first-line (SPP1) was 18.69 mo. Interestingly, differences in outcome were noticed according to immunophenotype. The best prognosis was observed in luminal A disease (OS= 46.72 mo, PFS1= 15.61 mo, SPP1= 25.43 mo). In luminal B disease, median OS was 27.66 mo, PFS1 8.94 mo, and SPP1 13.21 mo, respectively. In patients with HER2-positive disease, mainly treated with anti-HER2 therapy, outcome approached that of luminal A disease (OS= 41.10 mo, PFS= 9.89 mo, SPP1 18.69 mo). Patients with triple negative disease (TNBC) experienced the worst prognosis (OS= 8.54 mo, PFS1= 4.04 mo, SPP1= 2.83 mo). Conclusions: The study demonstrated different results in the measures of outcome according to the line of treatment and specific disease characteristics. As a consequence, endpoints of clinical trials should be tailored taking into consideration prognostic/predictive factors as well as the line of treatment.
Phase Ib study of TSPP (thymidilate synthase poly-epitope peptide) vaccine in combination with GOLFIG chemoimmuno-burst in advanced colon cancer patients: Results from the TSPP/VAC-1/c trial.
Authors: Correale Pierpaolo , Pastina Pierpaolo , Botta Cirino , Rossetti Maria Grazia , Apollinari Serena , Bestoso Elena , Fioravanti Antonella , Guidelli Giacomo , Chellini Federico , Mantovani Giovanni , Gori Savellini Gianni , Carbone Salvatore Francesco , Ricci Veronica , Basile Assunta , Pagliuchi Marco , Tagliaferri Pierosandro , Pirtoli Luigi , Cusi Maria Grazia
Published: March 2020

e13098

Background: TSPP is a poly-epitope peptide anticancer vaccine able to generate a thymidylate syntase (TS) specific T cell response with antitumor activity in preclinical models, expecially when used in sequential combination with 5-FU. TS is in fact involved in DNA replication and is the major pharmacological target of fluoropyrimidines whose activity leads in turn to TS upregulation. On these basis we designed a phase Ib trial where we investigated the safety and the immunobiological activity of TSPP + GOLFIG (Gemcitabine + FOLFOX-4 + GM-CSF + IL-2) regimen. Methods: TSPP/VAC-1 (Eudract 2009-016897-33) is a monocentric dose finding Phase Ib trial. 12 pretreated metastatic colon cancer patients were enrolled between April and November 2011. All of them received biochemotherapy according to the GOLFIG regimen biweekly and TSPP (on day 7) diluted in motanide ISA720 (1:1) at the dosage of 100 (3 patients), 200 (3 patients) and 300 ug (6 patients). Results: No life-threatening adverse events were observed. Grade 3 leucopenia (2 cases), thrombocitopenia (3 cases), anemia (3 cases), nausea/vomiting (2 cases), and dyarrhea (4 cases) along with 3 cases of oxaliplatin reaction were recorded. Fever and flu-like symptoms were common along cytokine administration. An IFN-gamma ELISPOT assay recorded an increase in TS specific CTL precursors in 3/6 evaluable patients; it was also observed an increase in lymphocytes, eosinophils, terminal effector memory and central memory T cells and in both c-ANCA and p-ANCA (P= 0.02) in the peripheral blood and a significant reduction in inflammatory markers (LDH, CRP and ESR). Quality of life analysis revealed a general improvement in patients’health state along the treatment. 1 partial response, 6 disease stabilizations and 2 progessions of disease were recorded. TSPP Maximal Tolerated Dose was not reached, while the Optimal Biological Dose was identified at 300 ug. Conclusions: Our results suggest that the GOLFIG + TSPP vaccine combination is safe and immunologically active and deserves to be evaluated in a further phase II trial.
Interesting possibilities to improve the safety of ipilimumab.
Authors: Bakacs Tibor , Moss Ralph W , Mehrishi Jitendra , Szabo Miklos
Published: March 2020

e13102

Background: The problem of the ipilimumab therapy is that a common molecular target (CTLA-4 receptor) is expressed on the targeted and non-targeted T cells, respectively. We suggest that the frequency of immune-related adverse events can be reduced by treating conditions in which the number of the targeted cells is larger than the non-targeted ones and/or by improving the accuracy of the targeting. Methods: The first approach was tested in a phase I clinical trial in patients with relapsed malignancy following allogeneic hematopoietic stem cell transplantation. An ipilimumab blockade was used against CTLA-4 positive allogeneic T cells (responding patients had greater than 80% donor T-cell chimerism). The graft-versus-malignancy (GVM) effects were augmented without a significant impact on graft-versus-host disease (GVHD). A feasible approach could also be pretargeting that has been worked out for radioimmunodetection and radioimmunotherapy. To this end we suggest a testable thought experiment for the treatment of chronic lymphocytic leukemia (CLL). First a streptavidin (StAv) modified anti-CD19 mAb is administered, which is followed by the subsequent delivery of biotin labeled anti-CTLA-4 mAb. The latter endows T cells with the ability to travel to tumor sites without prematurely succumbing to apoptosis, while streptavidin’s ultra-high affinity for biotin (10-15M) ensures a tight bond to the biotin labeled anti-CTLA-4. Results: Using the law of mass action we calculated that above 1 mg/L ipilimumab concentration (i.e. about 5 mg/patient ~70kg) more than 80% of anti-CD19-StAv sensitized B cells will have been bound to anti-CTLA-4-biotin sensitized T cells. Conclusions: This way, the forces of the immune system liberated by the anti-CTLA-4 antibody blockade could be focused with laser sharp accuracy on CLL cells without collateral damage to normal host cells. One should note that 0.3 mg/kg ipilimumab alone already caused mild autoimmunity.
Autologous heterotopic transplant of peripheral blood associated to chemotherapy as immunotherapy of solid tumors.
Authors: Lasalvia-Prisco Eduardo M. , Goldschmidt Pablo , Galmarini Felipe , Vazquez Jesus , Lasalvia-Galante Eduardo E. , Golomar Wilson , Cucchi Silvia
Published: March 2020

e13105

Background: Successive developed configurations of Autologous Heterotopic Transplant of Peripheral Blood (AHTOPB) were reported lately. In advanced stages of different primary tumors, AHTOPB has improved the antitumoral effect of chemotherapy (CH). A cohort of patients (pts) was selected from these trials to explore the biological response modifications (angiogenesis, immunity) associated to such antitumoral effect. Methods: From 6 reported 2-arm trials (CH and AHTOPB+CH treated), 60 pts were selected, 5 from each arm and trial. Trials referred to Breast, NSC Lung, prostate, ovary, pancreas, colon. Inclusion criteria: histology confirmed, M1 stage, unresectable, measurable according Response Evaluation Criteria in Solid Tumors (RECIST), PS 0-2 according Eastern Cooperative Oncology Group (ECOG), not previous CH, organic functions preserved. Exclusion criteria: brain M1, treated comorbility, additional antitumor treatment. AHTOPB procedure as it was described: 1) Cyclophosphamide low dose, 2) Immunization site conditioning with activated autologous blood, 3) Immunization with tumor antigens released to the blood inside heat shock proteins. CH: standard first line recommended for each tumor. Assessments. In venous blood sample pre/post treatment: T-Regulatory cells (T-Reg); Activated Dendritic cells (aDC); Lymphocyte Proliferative Index AHTOPB challenged (LPI); Angiostatin (AT); Interleukine-2 (IL2); Interleukine-12 (IL12); Gamma Interferon (G-If). In sentinel lymph-node of immunization site post AHTOPB treatment: T-Reg and aDC. In clinical follow up: 30-day Tumor Growth (TG) according RECIST; Disease control rate (DCR), Overall Survival (OS), Toxicities (Tx) according NCI (≥3). Measurements were expressed as % of pretreatment mean value in same primary tumor; % of 2 treatments were statistically compared using two tailed Student T-test. Results: Arm AHTOPB+CH, comparable to arm CH, evidenced significant decrease T-Reg, VEGF, TG, and increase aDC, LPI, AT, IL2, IL12, G-If, DCR. No significant different toxicity. Conclusions: In cancer pts, ATHOPB+CH elicits the conditioning to produce biological responses compatible with antitumor immunotherapy.
Tumor hypoxia as a mechanism of resistance to bevacizumab in a murine model.
Authors: Heijmen Linda , Boerman Otto C. , Punt Cornelis J. A. , Ter Voert E. , Oyen Wim J.G. , Bussink Jan , Sweep Fred C.G.J. , Laverman Peter , Span Paul , de Geus- Oei Lioe-Fee , Heerschap Arend , Van Laarhoven Hanneke W.M.
Published: March 2020

e13111

Background: Despite the promise of preclinical and early phase clinical studies, the efficacy of bevacizumab in solid tumors is more limited than expected. One of the presumed reasons is the induction of tumor hypoxia by the anti-angiogenic effects of bevacizumab, leading to therapy resistance. The aim of this study was to assess the effect of bevacizumab on tumor hypoxia in vivo in a colorectal cancer model, using functional imaging techniques. Methods: Nude mice with s.c. LS174T colon carcinoma xenografts (0.05 - 0.3 cm3) were treated with bevacizumab (5 mg/kg; 2/wk, i.p.) or saline as a control. To assess tumor hypoxia in vivo 18F-MISO-PET microPET or T2*-MRI images were acquired of separate groups of mice (n=5) before treatment and at day 2, 6 and 10 days after start of treatment. Tumors were harvested directly after imaging to microscopically assess the hypoxic fraction (pimonidazole staining) and vascular density (9F1 staining). Results: Linear regression analyses showed that FMISO uptake increased significantly more over time in the control group than in the bevacizumab group (beta -0.44, p=0.02), indicating that bevacizumab reduced the inherent increase in tumor hypoxia over time. T2* time increased significantly less in the bevacizumab group (beta -0.45, p=0.01), indicating a higher deoxyhemoglobine concentration, which might indicate a higher perfusion of the tumor and thus less hypoxia. The hypoxic fraction did not change over time and no difference was observed between the tumors in the treated and the control group. Vessel density significantly decreased over time in the bevacizumab group (beta -0.25, p=0.05), while the hypoxic fraction remained unchanged in the control group. Conclusions: The bevacizumab-induced changes in the tumor were most prominent at 10 days after treatment initiation, implying a build-up effect of repeated bevacizumab administration. The treatment induced changes could be detected with both T2*-MRI as well as with FMISO microPET. Bevacizumab did not induce tumor hypoxia, despite the observed decrease in vascular density. Therefore, induction of tumor hypoxia as a resistance mechanism to bevacizumab treatment seems unlikely.
Intensified loading schedule of trastuzumab with concurrent taxane: Pilot study of cardiotoxicity and safety monitoring.
Authors: Zvirinska Lenka
Published: March 2020

e13112

Background: Recent update of NCCGT trial (Perez et al. J Clin Oncol. 2011;29:4491-4497) has shown only minor trend toward improvement in DFS with concurrent administration of chemotherapy with Transtuzumab. However data on pharmacokinetics of trastuzumab (Leyland-Jones et al. J Clin Oncol. 2010;28:960-966) suggest that more intense loading schedule could achieve effective levels earlier which might be vital for optimal effect. Methods: Therefore we have analysed data of patient cohort treated in single institution (UHCW), in the first instance monitoring difference in cardiotoxicity in patients treated by standard concurrent trastuzumab + taxane regime (8mg/kg followed by 3 weekly 6mg/kg) compared to patients treated with slightly intensified trastuzumab regime (8 mg/kg followed by 6 mg/kg a week later and continue starting with standard 3 weekly 6 mg/kg dose). Results: Altogether 34 patients have been identified starting treatment with trastuzumab in UHCW between October 2008 and October 2010. Median follow up was 12 months (8- 48). 20 were treated by standard concurrent regime, 14 by intensified. 30 patients received 300 – 400mg/m2 of epirubicin prior to trastuzumab. EF was monitored by MUGA every 3 months as per NICE recommendations at the time.
Trial of anti-CD3 x anti-EGFR bispecific antibody (EGFRBi) in patients with EGFR-expressing recurrent/metastatic non-small cell lung cancer (NSCLC) and solid tumors.
Authors: Rathore Bharti , Davol Pam , Rathore Ritesh , Cummings Francis , Birnbaum Ariel E. , Maizel Abby , Lum Lawrence G
Published: March 2020

e13124

Background: Patients (pts) with advanced NSCLC and solid tumors have a poor prognosis and novel approaches to improve targeting and cytotoxicity directed at these cancers are needed. Anti-CD3 activated T cells (ATC) can be expanded and armed with anti-CD3 x anti-EGFR bispecific antibody (EGFRBi). Methods: In a phase I trial using ATC armed with EGFRBi, 6 pts (NSCLC = 5 pts, pancreatic cancer =1 pt) were treated with 8 infusions (twice/week) for 4 weeks with 5, 10 billion armed ATC/dose to determine the maximum tolerated dose (3 pts each). T cells from leukopheresis were activated and expanded with anti-CD3/IL-2 for 14 days, harvested, armed with EGFRBi and cryopreserved. IL-2 (3 x 105 IU/m2/daily) and GM-CSF (250 µg/m2 twice/week) was given beginning 3 days prior to the 1st infusion and ending++ 3, performance status (PS 0= 2 pts, PS 1= 4 pts). Results: Restaging scans revealed stable disease in 2 pts and progression in 4 pts. One pt with NSCLC at each dose level had stable disease (23 mths, 11 mths). ATC infusions were well tolerated and noticeable toxicities included chills (grade 2= 1 pt, grade 3= 1 pt); headache (grade 1= 2 pts, grade 2= 2 pts), dizziness (grade 2= 1 pt), nausea (grade 1= 2 pts). Accrual continues for dose levels at 20, 40 billion armed ATC per dose. No major toxicities with IL-2 were observed; bone pain resulted in dose reductions for GM-CSF in 2 pts. Conclusions: Infusions of EGFRBi armed ATC can be safely accomplished in patients with advanced NSCLC and solid tumors; accrual continues.
New approach to adjuvant therapy in high-risk melanoma patients.
Authors: Elias E. George
Published: March 2020

e13129

Background: Cutaneous melanoma is relatively chemoresistant, but it is an immunogenic tumor. However, it is very heterogenetic. As a result, the present adjuvant therapy has been met with limited success due to non-specifity of therapy. GM-CSF administration in cutaneous metastatic melanoma increases the number and activation of naive dendritic cells which are antigen presenting cells that can process tumor antigens, while IL-2 stimulates and activates mainly T cells. Hypothesis: The consequential administration of a single short course of both agents intradermally at the primary site, prior to its excision, can result in high tumor cell kill and forms a triad of autologous tumor cells, dendritic cells and and T lymphocytes. This triad can be capable of inducing cytotoxic cells that can be taken up by the lymphatics to regional lymph nodes (LN). Methods: Newly diagnosed high risk patients, each is to receive a single injection of GM-CSF: 500 microgram at the primary (biopsy) site on day # 1, followed by IL-2: 11 million IU/day injected at the same site on days 2 and 3, one week before the definitive surgery. Postoperatively, the patient is to receive low dose GM-CSF at 250 microgram twice/week, self-administered subcutaneously, to maintain the initial response. Results: In a pilot study, the pathological examination of rsected tissues from treated patient revealed complete tumor necrosis with massive histiocytosis at the primary site. Immunohistochemical studies showed overexpression of CD4+, CD8+, CD25+ and CD83+ both at the primary and LN. Conclusions: This non-toxic outpatient program is safe and well tolerated. It can initiate patient-specific and tumor-specific cytotoxic T cells that can overcome heterogeneity of the tumor and confers therapeutic specificity. It also establishes the lymphatic route of administration. Postoperative subcutaneous administration of GM-CSF is to systemically maintain the initial response. The beneficial effect of this approach on survival has to be evaluated in a prospective controlled randomized clinical trial.
Signaling pathway differences between EGFR TKI-sensitive and -resistant non-small cell lung cancer cells.
Authors: Fei Shi-Jiang , Zhang Xu-Chao , Dong Song , Cheng Hua , Zhang Yi-Fang , Chen Zhihong , Xie Zhi , Wu Yi Long
Published: March 2020

e13510

Background: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have showed dramatic clinical benefits in advanced non-small cell lung cancer (NSCLC), but the biggest problem is resistance. How to overcome resistance is still a serious problem to clinical practice. The aims of this studies was to analyse the signaling pathway differences between the EGFR TKI-sensitive and resistant NSCLC cell lines and to seek potential molecular targets for tumor treatment after resistance. Methods: We selected four types of NSCLC cell lines: PC-9(exon19 del), PC9GR (gefitinib-acquired resistant cell), H1650(exon19 del), H1975 (L858R+T790M), signaling pathway differences between PI3K/Akt/mTOR,MAPK and STAT3 were analysed through western blot, immunoprecipitation and confocal microscope. Results: Activation status of EGFR super signaling pathways: PI3K-Akt, Erk and STAT3 between EGFR TKI- sensitive and resistant NSCLC cells was different under basic state. The phosphorylation levels of Akt ser473, FOXO1 and Erk in EGFR TKI-resistant cells, especially in H1650 cell, were more higher than that in PC9 cell except for p-STAT3. p-STAT3 expression in PC9 was higher than that in resistant NSCLC cells. The further analysis showed that cellular localizations of p-STAT3 between sensitive and resistant cells were different through confocal microscope, p-STAT3 in PC9 and H1975 cells was almost in nucleus, while in PC9GR and H1650 cells was almost in cytoplasm. The assay of mTORC2 kinase activities in vitro showed that mTORC2 kinase activities in the resistant cells were obviously higher than that in sensitive cells. Conclusions: we found that EGFR TKI-sensitive and resistant NSCLC cells had different activation status of signaling pathways especially mTOR associated signaling pathways and different mTORC2 kinase activities. These results indicated that targeting mTOR with specific mTOR inhibitors would be a good strategy for NSCLC treatment after EGFR TKI resistance.
Rational for targeted polytherapy in sarcoma: Get the genie out of the bottle.
Authors: Dumont Sarah N. , Yang Dan , Dumont Amaury Gerard , Blay Jean-Yves , Trent Jonathan C.
Published: March 2020

e13519

Background: While a change of paradigm occurred in the last decade from chemotherapy to targeted therapy for cancer treatment, this work investigates the optimal combination of targeted agents with doxorubicin in sarcoma. Methods: Three sarcoma cell lines were studied RD18 (rhabdomyosarcoma), A204 (undifferentiated sarcoma) and TC 71 (Ewing's sarcoma) and exposed to increasing concentrations of vorinostat (HDAC inhibitor), 17-DMAG (Hsp90 inhibitor), abacavir (anti-telomerase) and sorafenib (tyrosine kinase inhibitor) alone, combined 2 by 2, then with doxorubicin. Viability was assessed by MTS assay. The Chou and Talalay combination index (CI) was used to determine additive (CI=1), synergistic (CI<1) or antagonistic effect (CI>1). Cell cycle analysis, measure of apoptosis by Annexin V and caspase 3/7 activity were studied using flow cytometry analysis and luminescent assay. Results: In monotherapy, the agents showed 30% to 90% decrease in viability but abacavir, which remained less active. Combination therapies with vorinostat, sorafenib and 17-DMAG showed strong synergism. Abacavir was found antagonistic with each drug. Either vorinostat or 17DMAG synergized with doxorubicin, achieving 60% cell killing compared to doxorubicin alone 12% (p=0.007). However, no synergy was observed for sorafenib with doxorubicin. The triple therapy vorinostat, 17DMAG and Doxorubicin did not show synergism but transiently increased the subG1 population at 24H, 70% compared to 30% in monotherapy with an increase in early caspase-independent apoptosis (11% to 36%, p= 0.0008). Conclusions: This work provides evidence of synergism of dual combinations of vorinostat, 17DMAG and sorafenib. In adjunction to doxorubicin, these combinations enhance doxorubicin cytotoxicity at therapeutically relevant concentrations. [Table: see text]
Evaluation of dosimetric variance in whole breast forward intensity modulated radiotherapy based on 4D CT and 3D CT.
Authors: Wang Wei , Li Jianbin
Published: March 2020

e13530

Background: To explore and compare the dosimetric variance in forward intensity modulated radiotherapy (IMRT) based on 4D CT and 3D CT after breast conserving surgery. Methods: Seventeen patients after breast conserving surgery underwent the 3D CT simulation scans followed by respiration-synchronized 4D CT simulation scans on the state of free breathing. The treatment plan constructed using the end inspiration (EI) scan was then copied and applied to the end expiration (EE) and 3D scans and the dose distribution was calculated separately. Dose–volume histograms (DVHs) parameters for the CTV, PTV, ipsilateral lung (IPSL) and heart were evaluated and compared. Results: The CTV volume amplitude was 11.93 ± 28.64 cm3, and volume of the CTV receiving 95%, 100%, and 103% prescription dose among different scans were all differed by < 0.4%. Mean PTV dose at EE was lower than EI (t = 2.87, p = 0.011), but there were no statistice significance between 3D CT scan and EI, EE scans (t = 1.06, -1.59; p = 0.304, 0.132). The homogeneity index (HI) at EI, EE, 3D plans were 0.156 ± 0.02, 0.162 ± 0.02, 0.161 ± 0.02, respectively, and difference only between EI and EE (t = -2.56, p = 0.021). The highest conformal index (CI) was at EI phase (t = 4.55, 2.70; p = 0.000, 0.016), and there was no significant difference between EE and 3D (t = 0.04, p = 0.967). The V20, V30, V40, V50 and Dmean of IPSL at EE phase were lower than EI (t = 2.39~5.54, p = 0.000~0.030). There were no significant differences in all the indexes for heart (t = -1.77~1.40, p = 0.128~0.693). Conclusions: The breast deformation during respiration may be disregarded in whole breast IMRT; PTV dose distribution was changed significantly between EI and EE phase, and the differentiation of the lung high dose area between EI and EE phase may induced by thorax expansion. 3D treatment planning is sufficient for whole breast forward IMRT, but 4D CT scans assist with respiratory gating ensure precise delivery of radiation dose.
Potential increase in androgen sensitivity following antisense oligonucleotide suppression of Bcl-2 in a prostate cancer model.
Authors: Guinan Patrick , Rubenstein Marvin , Hollowell Courtney M.P.
Published: March 2020

e13532

Background: Antisense oligonucleotides (oligos) have been employed against in vivo and in vitro prostate cancer models targeting growth stimulatory gene products. While most oligos have targeted growth factors or their receptors, others have been directed against inhibitors of apoptosis and mediators of androgen action. In LNCaP cells we evaluated a set of oligos which targeted and comparably suppressed the expression of the apoptosis inhibitor protein bcl-2. To evaluate the specificity of this type of gene therapy, targeted and non-targeted genes were evaluated for their expression. The purpose was to identify whether additional (non-targeted) genes altered their expression in a manner which could circumvent or compensate for the suppression of bcl-2, promoting growth through increased androgen sensitivity. Methods: LNCaP prostate cells were treated with mono- and bispecific oligos directed against bcl-2. Employing RT-PCR the expression of both targeted and non-targeted genes (androgen receptor [AR], coactivators p300 and creb binding protein [CREB]) was determined. Results: LNCaP cells adapted to the suppression of bcl-2 (an apoptosis inhibitor) with enhanced expression of both AR and p300. This suggests an increased sensitivity to androgens. Conclusions: In this model, oligo treatment directed against bcl-2, may be evaded through a compensatory increase in both AR and p300 expression. This promotion of tumor growth, and the altered expression of AR coactivators normally seen in advanced disease, suggests a tumor transition to a more aggressive phenotype following suppressive treatment directed against bcl-2.
An orthotopic xenograft model of metastatic nasopharyngeal carcinoma and its application in elucidating a therapeutic target that inhibits metastasis.
Authors: Smith Pamela Ann
Published: March 2020

e13536

Background: Nasopharyngeal carcinoma (NPC) is a serious health problem in many parts of the world. Although rare in the United States and Western Europe, it is the leading form of cancer in men between the ages of 20-44 years in parts of southeast Asia, and is prevalent in Indonesia, North Africa, the Middle East, amongst Eskimo populations, and represents one of the most frequent epithelial tumors of children in geographically higher risk areas. Current treatment protocols have resulted in good clinical outcomes if the disease is diagnosed and treated at an early stage. Unfortunately, metastatic NPC is refractory to treatment and carries an extremely poor prognosis. Particularly concerning is, that among head and neck cancers of epithelial origin, it is associated with the highest rate of metastasis. This highlights the need for models that are amenable to the study of metastatic NPC and adequate to test new potential therapies. Methods: In order to define and therapeutically target mechanisms that mediate nasopharyngeal carcinoma (NPC) metastasis, we have developed a unique orthotopic xenograft mouse model that accurately recapitulates the invasive and metastatic behavior of human disease. Results: Based on published clinical data and gene array analysis we performed on biopsy samples, we concluded that the PI3K/Akt/mTOR axis is involved in aggressive NPC tumor behavior. We thus chose it as a therapeutic target to test the utility of our orthotopic system for evaluating the effectiveness of a targeted treatment for metastatic NPC. We have subsequently demonstrated that both the development and growth of metastatic lesions are markedly reduced by the mTOR inhibitor sirolimus. Conclusions: Thus, this orthotopic model provides a platform to study potential therapeutics for advanced NPC and demonstrates that targeting the PI3K/Akt/mTOR pathway is a promising intervention against disseminated disease.
The impact of novel compound 2'-hydroxyflavanone on proteomic networks of VHL-mutant renal cell carcinoma.
Authors: Dalasanur Nagaprashantha Lokesh Prasad Gowda , Talamantes Tatjana , Singhal Jyotsana , Awasthi Sanjay , Singhal Sharad S. , Prokai Laszlo
Published: March 2020

e13557

Background: Von Hippel-Lindau (VHL) tumor suppressor is a major genetic factor that determines hypoxic acclimatization and response to therapy in renal cell carcinoma (RCC). Our initial studies revealed that 2'-Hydroxyflavanone (2HF) effectively inhibits the survival of VHL-mutant RCC. Hence, we performed proteomic analyses to study the signaling networks of clinical significance that are regulated by 2HF in VHL-mutant RCC. Methods: The 786-O cells, an established cell line from a male Caucasian patient, were provided by William Kaelin Jr., MD, Harvard Medical School, Boston, MA. The cells were cultured either in the presence or absence of 50 μM 2HF for 48 h in RPMI medium at 37°C in 5% CO2 and lysed in a buffer containing 6M urea. The lysate (200 µg of protein) was reduced, alkylated and digested with trypsin. Tryptic digests were desalted using a C-18 cartridge (Waters, USA) and subjected to liquid chromatography–tandem mass spectrometry (LC-MS/MS) analyses on linear ion trap–Fourier transform ion cyclotron resonance tandem mass spectrometer (LTQ-FT, Thermo) coupled to an Eksigent nano-LC system. MS/MS spectra were searched against human protein sequence database using Mascot (Matrix Science), and protein identifications were validated by Scaffold (version 3.0, Proteome Software). Quantifications were performed by extracted peptide intensities (Progenesis LC-MS, Nonlinear Dynamics). Results: 2HF treatment lead to up-regulation of 21 proteins including protein disulfide-isomerase A4 and macrophage migration inhibition factor. 2HF treatment lead to down-regulation of 23 proteins including vimentin, HSP90AB1, 14-3-3 gamma and glutaminase. The network analyses revealed that the identified proteins form critical nodes of regulation of vital tumor proteins like MAP3K14, ERK1/2, Myc and P53. Conclusions: Proteomic analyses of 2HF treated VHL-mutant RCC revealed a striking regulation of proteins of importance in RCC survival, invasion and metastases. The critical nodes of regulation as identified by the network analyses could facilitate further studies on the development of network targeting therapies using 2HF and other anti-cancer compounds for the effective management of VHL-mutant RCC.
Stromal and antitumor effects of BIBF 1120 (nintedanib) in a preclinical lung cancer model.
Authors: Gerber David E. , Kutluk Cenik Bercin , Ostapoff Katherine T , Brekken Rolf A.
Published: March 2020

e13563

Background: BIBF 1120 is an angiogenic receptor tyrosine kinase inhibitor that potently inhibits VEGFR, PDGFR and FGFR kinase activity in in vitro enzymatic assays. This study investigated the effect and mechanism of BIBF 1120, in vitro and in vivo, as a single agent and in combination with chemotherapy in preclinical models of lung cancer. Methods: Anti-tumor effects of BIBF 1120 in vitro were assessed using cell proliferation assays on five lung cancer lines (A549, Calu-3, Calu-6, H1993, H1703) with BIBF 1120 as a single agent and in combination with gemcitabine and cisplatin. To demonstrate anti-tumor activity in vivo, NOD/SCID mice bearing subcutaneous A549 grafts were treated daily with BIBF 1120 or BIBF 1120 plus chemotherapy. Perfusion studies were conducted using labeled dextrans. Ex vivo tumor tissues were assessed for architecture, microvessel density, pericyte coverage, proliferation, and apoptosis. Results: In vitro, BIBF 1120 did not show anti-proliferative effects at pharmacologically achievable concentrations (IC50>20μM) as a single agent; nor did it sensitize tumor cells to chemotherapy. However, BIBF 1120 inhibited primary tumor growth as a single agent and in combination therapy in subcutaneous endpoint studies with A549 xenografts. There was no significant difference in tumor architecture between BIBF 1120 and control-treated animals. Microvessel density (CD31,endomucin) and pericyte-covered vessels (NG2) were significantly decreased in BIBF 1120-treated animals compared to the control and chemotherapy only groups. Proliferation (phospho-histone 3) was decreased and apoptosis (cleaved caspase 3) was increased in BIBF 1120-treated animals compared to the control and chemotherapy only groups. Finally, BIBF 1120 significantly decreased perfusion in A549 xenografts. Conclusions: BIBF 1120 demonstrated potent anti-tumor and anti-angiogenic activity in a preclinical model of lung cancer. Because A549 was previously demonstrated to be an anti-VEGFR therapy resistant cell line, this study highlights the importance of FGFR and PDGFR pathways in the treatment of lung cancer.