Metastatic breast cancer patients treated with low-dose metronomic chemotherapy with cyclophosphamide and celecoxib: clinical outcomes and biomarkers of response

Background Preclinical results showing therapeutic effect and low toxicity of metronomic chemotherapy with cyclophosphamide (Cy) + celecoxib (Cel) for mammary tumors encouraged its translation to the clinic for treating advanced breast cancer patients (ABCP). Patients and methods A single-arm, mono-institutional, non-randomized, phase II, two-step clinical trial (approved by Bioethics Committee and Argentine Regulatory Authority) was designed. Patients received Cy (50 mg po.d) + Cel (200 mg p.o.bid). Patient eligibility criteria included: ABCP who progressed to anthracyclines, taxanes and capecitabine, ≤4 chemotherapy schemes, with good performance status. Several pro- and anti-angiogenic molecules and cells were determined as biomarkers. Informed consent was signed by all patients. Primary endpoint was clinical benefit (CB). Results Twenty patients were enrolled. Main clinical outcomes were prolonged disease stabilization and partial remission in 10/20 and 1/20 patients, respectively. CB was 55 %, and time to progression (TTP) was 21.1 weeks. Median TTP in patients who achieved CB was 35.6 weeks, and mean overall survival was 44.20 weeks. There were no grade 3/4 toxicities associated with treatment. Circulating endothelial cells (CECs) increased at the time of progression in patients who showed CB (P = 0.014). Baseline CECs and circulating endothelial progenitor cells showed marginal associations with TTP. Serum VEGF decreased (P = 0.050), sVEGFR-2 increased (P = 0.005) and VEGF/sVEGFR-2 ratio decreased during treatment (P = 0.041); baseline VEGF and VEGF/sVEGFR-2 were associated with TTP (P = 0.035 and P = 0.030, respectively), while sVEGFR-2 did not. Conclusions Treatment was effective, showing low toxicity profile and excellent tolerability. The combination had anti-angiogenic effect. Increased levels of CEC could be useful for detecting progression. Baseline VEGF and VEGF/sVEGFR-2 values could be useful as early predictors of response. Trial registration ANMAT#4596/09.Fil: Perroud, Herman A. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Alasino, Carlos María. Institute of Oncology of Rosario; ArgentinaFil: Rico, María José. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Mainetti, Leandro Ernesto. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; ArgentinaFil: Queralt, Francisco. Institute of Oncology of Rosario; ArgentinaFil: Pezzotto, Stella Maris. Research Council of the National University of Rosario (CIUNR); ArgentinaFil: Rozados, Viviana R. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Scharovsky, O. Graciela. Universidad Nacional de Rosario. Consejo de Investigaciones UNR, Facultad de Ciencias Médicas; Argentin

Nonclassical roles for IFN-γand IL-10 in a murine model of immunoedition

Aims: to characterize, by means of univariate and multivariate approaches, the Th1 and Th2 responses during the different phases of tumor immunoediting. Materials & Methods: we used a multivariate principal component analysis applied to analyze the joint behavior of serum concentrations of IFN-γ, IL-2, IL-10 and IL-4, during the different phases of tumor immunoediting, in CBi/L mice challenged with M-406 mammary adenocarcinoma. Results & Conclusions: Animals in equilibrium phase showed the widest variations in values of the four cytokines. In this experimental model the role of IFN-γ would be related to tumor growth and progression, while IL-10 would participate in the antitumor immune response. Lay abstract: Breast cancer is a complex, multifactor disease that affects about 10% of women in industrialized countries. The immune system has the ability to monitor the appearance of tumors, but the tumors have the ability to escape such rejection. For this reason, in order to design different therapeutic strategies, it is important to know the different mechanisms that take place when a tumor grows or when it is rejected. Here we sought to elucidate some of these mechanismsFil: del Giúdice, Antonela. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Pagura, Lucas. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Capitani, María Celeste. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; ArgentinaFil: Mainetti, Leandro Ernesto. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Scharovsky, Olga Graciela. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; Argentina. Universidad Nacional de Rosario. Consejo de Investigaciones de la Universidad de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Di Masso, Ricardo Jose. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; Argentina. Universidad Nacional de Rosario. Consejo de Investigaciones de la Universidad de Rosario; ArgentinaFil: Rico, Maria Jose. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Rozados, Viviana Rosa. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genética Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentin

Losartan improves the therapeutic effect of metronomic cyclophosphamide in triple negative mammary cancer models

Metronomic chemotherapy refers to the minimum biologically effective doses of a chemotherapy agent given as a continuous regimen without extended rest periods. Drug repurposing is defined as the use of an already known drug for a new medical indication, different from the original one. In oncology the combination of these two therapeutic approaches is called “Metronomics”. The aim of this work is to evaluate the therapeutic effect of cyclophosphamide in a metronomic schedule in combination with the repurposed drug losartan in two genetically different mice models of triple negative breast cancer. Our findings showed that adding losartan to metronomic cyclophosphamide significantly improved the therapeutic outcome. In both models the combined treatment increased the mice's survival without sings of toxicity. Moreover, we elucidated some of the mechanisms of action involved, which include a decrease of intratumor hypoxia, stimulation of the immune response and remodeling of the tumor microenvironment. The remarkable therapeutic effect, the lack of toxicity, the low cost of the drugs and its oral administration, strongly suggest its translation to the clinical setting in the near future.Fil: Mainetti, Leandro Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Rico, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Kaufman, Cintia Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Grillo, Monica Carolina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Guercetti, Julian. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Baglioni, María Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: del Giúdice, Antonela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Capitani, María Celeste. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Fusini, Matías. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Rozados, Viviana Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; ArgentinaFil: Scharovsky, Olga Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Medicas. Instituto de Genetica Experimental; Argentina. Metronomics Global Health Initiative; Franci

Comparative Effectiveness of Two Metronomic Chemotherapy Schedules. Our Experience in the Preclinical Field

Metronomic chemotherapy refers to the chronic, equally spaced, delivery of low doses of chemotherapeutic drugs, without extended interruptions. Previously, we developed two combined metronomic schemes for the treatment of murine mammary tumors. The aim of this study was to compare their effects on tumor and metastasis growth, survival, and toxicity. Metronomic chemotherapy with Cyclophosphamide + Celecoxib (Cy + Cel) showed higher antimetastatic power than Cyclophosphamide + Doxorubicin (Cy + Dox), while being similar in other aspects. That difference, plus the advantage that represents its oral administration, suggests that the Cy + Cel combination is more suitable than Cy + Dox for metronomic chemotherapy of mammary tumors and could be proposed to the translation to the clinic.Fil: Rico, María José. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Perroud, Herman A. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Mainetti, Leandro Ernesto. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; ArgentinaFil: Rozados, Viviana R. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Scharovsky, Graciela Olga. Universidad Nacional de Rosario. Consejo de Investigaciones UNR, Facultad de Ciencias Médicas; Argentin

Regulatory T cells but not NKT I cells are modulated by a single low-dose cyclophosphamide in a B cell lymphoma tumor-model

Aim: Experimental and clinical studies showed that the administration of cyclophosphamide (Cy) in low doses leads to an enhancement of the antitumor immune response. Our objective was to study the modulation, if any, by low dose Cy, of T regulatory (Treg) and natural killer T (NKT) I cells, two cell populations of the innate immune response with opposing effects on the tumors, in a rat B cell lymphoma model. Methods: Inbred e rats were challenged s.c. with L-TACB lymphoma and on day 14 animals were distributed in two groups. Treated: injected i.p. with cyclophosphamide (10mg/kg of body weight) and Control: injected i.p. with saline. Blood samples were taken from days 0 to 21 and the percentage of T regulatory and natural killer T I cells were determined by flow cytometry. Results: We found that the increase of natural and inducible T regulatory cells of peripheral blood achieved during tumor growth was significantly downregulated by cyclophosphamide. On the contrary, natural killer T I cells were not modulated by the treatment. Conclusion: The antimetastatic effect of a single low dose of Cy would be due, at least in part, to downregulation of natural and inducible T regulatory cells

Inmunomodulación y antiangiogénesis en la terapéutica oncológica. De la investigación básica a la clínica

La investigación básica y pre-clínica en oncología celular y molecular son pilares fundamentales en los que se apoyan la mayoría de los adelantos en la terapéutica del cáncer. Los hallazgos obtenidos y su aplicación en la práctica clínica constituyen la causa del avance sostenido en el tratamiento de la enfermedad neoplásica. El objetivo de este trabajo es resumir y discutir los resultados pre-clínicos en inmunomodulación y anti-angiogénesis para el tratamiento de diversos tipos de tumores, obtenidos en nuestro Instituto durante los últimos 15 años, y la posterior traslación y aplicación del conocimiento experimental en un Ensayo Clínico Fase I/II. Se describen los resultados que contribuyeron a descifrar los mecanismos de acción de la inmunomodulación antimetastásica con ciclofosfamida, la quimioterapia metronómica con diferentes drogas únicas o combinaciones, y finalmente el diseño y resultados preliminares de un ensayo clínico de quimioterapia metronómica para pacientes con cáncer de mama avanzado.Immunomodulation and antiangiogenesis in cancer therapy. From basic to clinical research. Basic and pre-clinic research in cellular and molecular oncology are the main supports accounting for the advancement in cancer therapeutics. The findings achieved, and their implementation in clinical practice are responsible for the permanent improvement in the treatment of the neoplastic disease. Our present objective is to summarize and discuss the pre-clinical findings in immunomodulation and anti-angiogenesis for the treatment of several types of tumors obtained in our Institute during the last 15 years, and the subsequent translation and application of the acquired experimental knowledge in a Phase I/II Clinical Trial. We present the results and mechanisms of action of antimetastatic immunomodulation with cyclophosphamide, the metronomic chemotherapy with different single drugs and their combinations, and finally the design and preliminary results of a clinical trial with metronomic chemotherapy for patients with advanced breast cancer

Achievements and challenges in the use of metronomics for the treatment of breast cancer

Two interesting therapeutic proposals for cancer treatment emerged at the beginning of the 21st century. The first one was metronomic chemotherapy, which refers to the chronic administration of chemotherapeutic agents, in low doses, without extended drug-free periods. Then, the idea of drug repositioning in oncology, the use of well-known drugs that were created for other uses to be utilized in oncology, gained strength. Shortly after, the two strategies were merged in one, named metronomics. Both approaches share several features which make metronomics an appealing choice for cancer treatment: use of known and approved drugs, thus diminishing the time necessary to enter to the clinic, therapeutic effect, low toxicity, oral administration, better life quality, low costs because of the use of, generally, out of patent drugs, possibility of use, even in countries with very low economic resources. Many chemotherapy and repurposed drugs were tested with metronomics approaches for the treatment of mammary cancer, the most common malignancy in women worldwide, leading to high rates of mortality. The wide range of therapeutic models studied, paralleled the wide range of responses obtained, like tumor growth and metastasis inhibition, overall survival increase, lack of toxicity, better life quality, among others. The accomplishments reached, and the challenges faced by researchers, are discussed.Fil: Scharovsky, O. Graciela. Universidad Nacional de Rosario. Consejo de Investigaciones UNR, Facultad de Ciencias Médicas; ArgentinaFil: Rico, María José. Institute of Experimental Genetics.School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Mainetti, Leandro Ernesto. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Perroud, Herman A. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; ArgentinaFil: Rozados, Viviana R. Universidad Nacional de Rosario. Facultad de Ciencias Médicas; Argentin

Safety and therapeutic effect of metronomic chemotherapy with cyclophosphamide and celecoxib in advanced breast cancer patients

Metronomic chemotherapy (MCT), the chronic administration, at regular intervals, of low doses of chemotherapeutic drugs, without extended rest periods, allows chronic treatment with therapeutic efficacy and low toxicity. Our pre-clinical results suggested that combined MCT with cyclophosphamide (CY) and celecoxib (CEL) could inhibit breast cancer growth. The aim of this study was to determine the toxicity, safety and efficacy of the oral chronic administration of CY 50 mg p.o. daily, plus CEL 400 mg (200 mg p.o. bid) in advanced breast cancer patients (ABCP). The therapeutic response consisted in stable disease in a high proportion of patients and one partial response. The overall clinical benefit rate obtained was 66.7%. Toxicity was low. Evaluation of quality of life showed no changes during the response period. MCT with CY + Cel is safe and shows therapeutic effect in ABCP.Fil: Perroud, Herman A. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Rico, María José. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Alasino, Carlos María. Institute of Oncology of Rosario; ArgentinaFil: Queralt, Francisco. Institute of Oncology of Rosario; ArgentinaFil: Pezzotto, Stella Maris. Research Council of the National University of Rosario (CIUNR); ArgentinaFil: Rozados, Viviana R. Institute of Experimental Genetics. School of Medical Sciences. National University of Rosario; Rosario; ArgentinaFil: Scharovsky, O. Graciela. Universidad Nacional de Rosario. Consejo de Investigaciones UNR, Facultad de Ciencias Médicas; Argentin

The Immune Response and the Therapeutic Effect of Metronomic Chemotherapy With Cyclophosphamide

Metronomic chemotherapy (MCT) is a novel therapeutic strategy for cancer treatment endowed with an antiangiogenic effect. It refers to regular administration of low doses of cytotoxic drugs, with minimal or no drug-free breaks. Previously, we demonstrated the immunomodulating activity of a single low-dose of cyclophosphamide (Cy) and the antitumor effect of MCT with Cy on established rat lymphomas and sarcomas. Here, we examined whether the immune response is responsible for the antitumor effect of MCT with Cy on L-TACB lymphoma. Inbred e rats and nude mice were subcutaneously challenged with L-TACB. After 7 days, they were distributed into two experimental groups: 1) treated animals, which were injected IP with Cy (10 mg/kg body weight) three times per week, and 2) control animals, which received IP saline injections. Exponential growth and decay and tumor doubling time were calculated. Also, serum IL-10 levels were measured. One hundred percent of treated rats showed tumor regression versus 0% of control rats. The increase of tumor-induced IL-10 levels was reverted by the treatment with Cy. On the other hand, there were no tumor regressions, in treated or control nude mice. However, the tumor doubling times of treated nude mice were significantly higher than those of control mice, implying that other antitumor mechanism(s), independent of the adaptive immune response, might be taking place. Our present results indicate that modulation of the immune response would be involved in the antitumor effect of MCT with Cy, because the absence of the specific immune response impairs, at least in part, its therapeutic effect in a lymphoma tumor model