Synergistic DNA-damaging effect in multiple myeloma with the combination of zalypsis, bortezomib and dexamethasone.

Despite new advances in multiple myeloma treatment and the consequent improvement in overall survival, most patients relapse or become refractory to treatment. This suggests that new molecules and combinations that may further inhibit important survival pathways for these tumor cells are needed. In this context, zalypsis is a novel compound, derived from marine organisms, with a powerful preclinical anti-myeloma effect based on the sensitivity of malignant plasma cells to DNA-damage induction; and it has already been tested in a phase I/II clinical trial in multiple myeloma. We hypothesized that the addition of this compound to the combination of bortezomib plus dexamethasone may improve efficacy with acceptable toxicity. The triple combination demonstrated strong synergy and higher efficacy compared with double combinations; not only in vitro, but also ex vivo and, especially, in in vivo experiments. The triple combination triggers cell death, mainly through a synergistic induction of DNA damage and a decrease in the nuclear localization of nuclear factor kappa B. Our findings support the clinical evaluation of this combination for relapsed and refractory myeloma patients.

The kinesin spindle protein inhibitor filanesib enhances the activity of pomalidomide and dexamethasone in multiple myeloma.

Kinesin spindle protein inhibition is known to be an effective therapeutic approach in several malignancies. Filanesib (ARRY-520), an inhibitor of this protein, has demonstrated activity in heavily pre-treated multiple myeloma patients. The aim of the work herein was to investigate the activity of filanesib in combination with pomalidomide plus dexamethasone backbone, and the mechanisms underlying the potential synergistic effect. The ability of filanesib to enhance the activity of pomalidomide plus dexamethasone was studied in several in vitro and in vivo models. Mechanisms of this synergistic combination were dissected by gene expression profiling, immunostaining, cell cycle and short interfering ribonucleic acid studies. Filanesib showed in vitro, ex vivo, and in vivo synergy with pomalidomide plus dexamethasone treatment. Importantly, the in vivo synergy observed in this combination was more evident in large, highly proliferative tumors, and was shown to be mediated by the impairment of mitosis transcriptional control, an increase in monopolar spindles, cell cycle arrest and the induction of apoptosis in cells in proliferative phases. In addition, the triple combination increased the activation of the proapoptotic protein BAX, which has previously been associated with sensitivity to filanesib, and could potentially be used as a predictive biomarker of response to this combination. Our results provide preclinical evidence for the potential benefit of the combination of filanesib with pomalidomide and dexamethasone, and supported the initiation of a recently activated trial being conducted by the Spanish Myeloma group which is investigating this combination in relapsed myeloma patients.

Effect of Oncoxin Oral Solution in HER2-Overexpressing Breast Cancer.

One of the most aggressive breast cancer subtypes includes tumors with high expression of HER2. Gene expression and functional studies have shown a link between HER2 overexpression and oxidative stress. Because of this, we hypothesized that Oncoxin Oral Solution (OOS), a composite product that contains several antioxidants, could have an antitumoral effect against HER2+ tumors. Dose-response studies, biochemical and cytometric assessment of the effect of OOS on cell cycle and apoptosis, and drug combination analyses were performed on BT474 and SKBR3 cells, 2 HER2-overexpressing breast cancer cell lines. OOS reduced the proliferation of these cells, and augmented the action of lapatinib, a HER2 inhibitor used in the breast cancer clinic. Moreover, OOS decreased growth of HER2+ tumors in mice. Mechanistically, OOS provoked cell cycle blockade through upregulation of p27 expression and downregulation of cyclin D levels. OOS also caused apoptotic cell death in HER2+ breast cancer cells, as indicated by increases in PARP cleavage as well as upregulation of caspase 8 and caspase 3 activities. These results demonstrate an antitumoral action of OOS in preclinical models of HER2+ breast cancer and suggest that it can be used with anti-HER2 therapies currently adopted as standard of care in the oncology clinic.

R-RAS2 overexpression in tumors of the human central nervous system.

Malignant tumors of the central nervous system (CNS) are the 10th most frequent cause of cancer mortality. Despite the strong malignancy of some such tumors, oncogenic mutations are rarely found in classic members of the RAS family of small GTPases. This raises the question as to whether other RAS family members may be affected in CNS tumors, excessively activating RAS pathways. The RAS-related subfamily of GTPases is that which is most closely related to classical Ras and it currently contains 3 members: RRAS, RRAS2 and RRAS3. While R-RAS and R-RAS2 are expressed ubiquitously, R-RAS3 expression is restricted to the CNS. Significantly, both wild type and mutated RRAS2 (also known as TC21) are overexpressed in human carcinomas of the oral cavity, esophagus, stomach, skin and breast, as well as in lymphomas. Hence, we analyzed the expression of R-RAS2 mRNA and protein in a wide variety of human CNS tumors and we found the R-RAS2 protein to be overexpressed in all of the 90 CNS cancer samples studied, including glioblastomas, astrocytomas and oligodendrogliomas. However, R-Ras2 was more strongly expressed in low grade (World Health Organization grades I-II) rather than high grade (grades III-IV) tumors, suggesting that R-RAS2 is overexpressed in the early stages of malignancy. Indeed, R-RAS2 overexpression was evident in pre-malignant hyperplasias, both at the mRNA and protein levels. Nevertheless, such dramatic changes in expression were not evident for the other two subfamily members, which implies that RRAS2 is the main factor triggering neural transformation.

A Case Series of Pregnant Women With Mitral Stenosis.

Pregnant women with mitral stenosis have a high cardiovascular risk, and their outcomes vary greatly. The management of these patients should be individualized and carried out by a multidisciplinary team. This case series presents 3 cases of pregnant women with severe mitral stenosis, describing their progression and therapeutic strategies.

Filanesib for the treatment of multiple myeloma.

Introduction: Kinesin spindle protein (KSP) is indispensable for the proper separation of spindle poles during mitosis. Importantly, this protein is expressed only in cells undergoing cell division and hence represents an appealing target for the treatment of cancer. Many KSP inhibitors have demonstrated a strong antitumoral effect in vitro, however, they have exhibited only limited activity in clinical trials. By contrast, the KSP inhibitor filanesib has demonstrated clinical efficacy in patients with multiple myeloma (MM).Areas covered: This article provides a comprehensive overview about the progress to date in the preclinical and clinical development of filanesib for the treatment of cancer, and particularly, MM.Expert opinion: Responses observed with filanesib alone or in combination with dexamethasone were encouraging in MM. However, the subsequent appearance of highly effective novel agents such as monoclonal antibodies, has hindered the development of agents such as filanesib that exhibit a more limited activity. Nevertheless, filanesib has shown interesting results for some patients when combined with carfilzomib and pomalidomide. Most importantly, the availability of a biomarker of response such as alpha 1-acid glycoprotein (AAG), could be key to the identification of patients that could benefit most from these combinations.

Protein Translation Inhibition is Involved in the Activity of the Pan-PIM Kinase Inhibitor PIM447 in Combination with Pomalidomide-Dexamethasone in Multiple Myeloma.

BACKGROUND: Proviral Insertion site for Moloney murine leukemia virus (PIM) kinases are overexpressed in hematologic malignancies, including multiple myeloma. Previous preclinical data from our group demonstrated the anti-myeloma effect of the pan-PIM kinase inhibitor PIM447. METHODS: Based on those data, we evaluate here, by in vitro and in vivo studies, the activity of the triple combination of PIM447 + pomalidomide + dexamethasone (PIM-Pd) in multiple myeloma. RESULTS: Our results show that the PIM-Pd combination exerts a potent anti-myeloma effect in vitro and in vivo, where it markedly delays tumor growth and prolongs survival of treated mice. Mechanism of action studies performed in vitro and on mice tumor samples suggest that the combination PIM-Pd inhibits protein translation processes through the convergent inhibition of c-Myc and mTORC1, which subsequently disrupts the function of eIF4E. Interestingly the MM pro-survival factor IRF4 is also downregulated after PIM-Pd treatment. As a whole, all these molecular changes would promote cell cycle arrest and deregulation of metabolic pathways, including glycolysis and lipid biosynthesis, leading to inhibition of myeloma cell proliferation. CONCLUSIONS: Altogether, our data support the clinical evaluation of the triple combination PIM-Pd for the treatment of patients with multiple myeloma.

The Novel Pan-PIM Kinase Inhibitor, PIM447, Displays Dual Antimyeloma and Bone-Protective Effects, and Potently Synergizes with Current Standards of Care.

PURPOSE: PIM kinases are a family of serine/threonine kinases recently proposed as therapeutic targets in oncology. In the present work, we have investigated the effects of the novel pan-PIM kinase inhibitor, PIM447, on myeloma cells and myeloma-associated bone disease using different preclinical models. EXPERIMENTAL DESIGN: In vitro/ex vivo cytotoxicity of PIM447 was evaluated on myeloma cell lines and patient samples. Synergistic combinations with standard treatments were analyzed with Calcusyn Software. PIM447 effects on bone cells were assessed on osteogenic and osteoclastogenic cultures. The mechanisms of PIM447 were explored by immunoblotting, qPCR, and immunofluorescence. A murine model of disseminated multiple myeloma was employed for in vivo studies. RESULTS: PIM447 is cytotoxic for myeloma cells due to cell-cycle disruption and induction of apoptosis mediated by a decrease in phospho-Bad (Ser112) and c-Myc levels and the inhibition of mTORC1 pathway. Importantly, PIM447 demonstrates a very strong synergy with different standard treatments such as bortezomib + dexamethasone (combination index, CI = 0.002), lenalidomide + dexamethasone (CI = 0.065), and pomalidomide + dexamethasone (CI = 0.077). PIM447 also inhibits in vitro osteoclast formation and resorption, downregulates key molecules involved in these processes, and partially disrupts the F-actin ring, while increasing osteoblast activity and mineralization. Finally, PIM447 significantly reduced the tumor burden and prevented tumor-associated bone loss in a disseminated murine model of human myeloma. CONCLUSIONS: Our results demonstrate dual antitumoral and bone-protective effects of PIM447. This fact, together with the very strong synergy exhibited with standard-of-care treatments, supports the future clinical development of this drug in multiple myeloma. Clin Cancer Res; 23(1); 225-38. ©2016 AACR.

Preclinical anti-myeloma activity of EDO-S101, a new bendamustine-derived molecule with added HDACi activity, through potent DNA damage induction and impairment of DNA repair.

BACKGROUND: Despite recent advances in the treatment of multiple myeloma (MM), the prognosis of most patients remains poor, and resistance to traditional and new drugs frequently occurs. EDO-S101 is a novel therapeutic agent conceived as the fusion of a histone deacetylase inhibitor radical to bendamustine, with the aim of potentiating its alkylating activity. METHODS: The efficacy of EDO-S101 was evaluated in vitro, ex vivo and in vivo, alone, and in combination with standard anti-myeloma agents. The underlying mechanisms of action were also evaluated on MM cell lines, patient samples, and different murine models. RESULTS: EDO-S101 displayed potent activity in vitro in MM cell lines (IC50 1.6-4.8 µM) and ex vivo in cells isolated from MM patients, which was higher than that of bendamustine and independent of the p53 status and previous melphalan resistance. This activity was confirmed in vivo, in a CB17-SCID murine plasmacytoma model and in de novo Vk*MYC mice, leading to a significant survival improvement in both models. In addition, EDO-S101 was the only drug with single-agent activity in the multidrug resistant Vk12653 murine model. Attending to its mechanism of action, the molecule showed both, a HDACi effect (demonstrated by α-tubulin and histone hyperacetylation) and a DNA-damaging effect (shown by an increase in γH2AX); the latter being again clearly more potent than that of bendamustine. Using a reporter plasmid integrated into the genome of some MM cell lines, we demonstrate that, apart from inducing a potent DNA damage, EDO-S101 specifically inhibited the double strand break repair by the homologous recombination pathway. Moreover, EDO-S101 treatment reduced the recruitment of repair proteins such as RAD51 to DNA-damage sites identified as γH2AX foci. Finally, EDO-S101 preclinically synergized with bortezomib, both in vitro and in vivo. CONCLUSION: These findings provide rationale for the clinical investigation of EDO-S101 in MM, either as a single agent or in combination with other anti-MM drugs, particularly proteasome inhibitors.

Antitumoral effect of Ocoxin on acute myeloid leukemia.

Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy whose incidence is growing in developed countries. In the relapse setting, very limited therapeutic options are available and in most cases only palliative care can be offered to patients. The effect of a composite formulation that contains several antioxidants, Ocoxin Oral solution (OOS), was tested in this condition. When analyzed in vitro, OOS exhibited anti-AML action that was both time and dose dependent. In vivo OOS induced a ralentization of tumor growth that was due to a decrease in cell proliferation. Such effect could, at least partially, be due to an increase in the cell cycle inhibitor p27, although other cell cycle proteins seemed to be altered. Besides, OOS induced an immunomodulatory effect through the induction of IL6. When tested in combination with other therapeutic agents normally used in the treatment of AML patients, OOS demonstrated a higher antiproliferative action, suggesting that it may be used in combination with those standard of care treatments to potentiate their antiproliferative action in the AML clinic.

[Physical training in chronic heart failure: pathophysiology and clinical evolution]. / Entrenamiento físico en la insuficiencia cardíaca crónica: fisiopatología y evolución clínica.

Chronic heart failure has become one of the main global health problems; 23 million people suffer from this disease worldwide and age of onset has varied considerably over the past five decades, coinciding with other co-morbidities as longevity in the population increases. Treatment of heart failure has also shown striking variations in recent years. Such is the case of the substitution of sympathomimetic drugs by beta-blocking agents, which primarily means a conceptual change in the pathophysiological interpretation of this syndrome. Incorporating to the treatment of heart failure drugs such as angiotensin converting enzyme inhibitors and angiotensin receptor blockers has meant a great step forward in the treatment of patients with this condition that significantly has decreased mortality and morbidity. The latest introduction of the drug identified as angiotensin receptor-neprilysin inhibitor (LCZ696), launched in August 2014 with an even greater reduction in mortality and morbidity of heart failure and fewer side effects, offers a valid hope in the treatment of this pathology. Training and physical activity is another area of treatment being completely reassessed. Pathophysiological aspects that link the practice of systematic physical exercise with heart failure and how they both relate to clinical outcomes, morbidity and mortality in the trained patient are reviewed in this paper.

La insuficiencia cardíaca crónica se ha convertido en uno de los mayores problemas de salud a nivel global; 23 millones de personas padecen esta patología en todo el mundo y su edad de aparición ha variado notablemente en los últimos cinco decenios, coincidiendo incluso con otras comorbilidades en la medida en que aumenta la longevidad en la población. El tratamiento de la insuficiencia cardíaca también ha mostrado variaciones impresionantes durante los últimos años. Tal es el caso del cambio en el tratamiento de las drogas simpaticomiméticas por los agentes betabloqueadores, que implica sobre todo un cambio conceptual en la interpretación fisiopatológica de este síndrome. La incorporación al tratamiento de la insuficiencia cardíaca de los fármacos inhibidores de la enzima convertidora de la angiotensina y de los bloqueadores de los receptores de la angiotensina ha significado un gran paso de avance en el tratamiento de pacientes con esta patología al disminuir significativamente su mortalidad y morbilidad. La más reciente introducción de la droga identificada como angiotensin receptor–neprilysin inhibitor (LCZ696), lanzada en agosto 2014, con una aun mayor disminución de la mortalidad y morbilidad de la insuficiencia cardíaca y menos efectos secundarios, ofrece una valedera esperanza para el tratamiento de esta patología. El entrenamiento y la actividad física es otra área de tratamiento que está siendo completamente revalorada. Se revisan aquí aspectos fisiopatológicos que vinculan la práctica del ejercicio físico sistemático con la insuficiencia cardíaca y cómo se relacionan con la evolución clínica, morbilidad y mortalidad del paciente entrenado.

Evaluación funcional de un programa de entrenamiento físico en pacientes infartados con disfunción sistólica severa del ventrículo izquierdo / Functional evaluation of a physical training programme in myocardial infarction patients with severe left ventricular systolic dysfunction

Resumen Objetivo: Evaluar la factibilidad y los efectos sobre la capacidad funcional de un programa de entrenamiento físico supervisado, aplicado en pacientes con disfunción sistólica severa del ventrículo izquierdo después de infarto agudo de miocardio. Métodos: Se estudiaron 37 pacientes, de ambos sexos y sin límites de edad, con diagnóstico de disfunción sistólica severa del ventrículo izquierdo, después de haber sufrido un infarto agudo de miocardio, que consecutivamente se incorporaron al programa ambulatorio del Centro de Rehabilitación del Instituto de Cardiología. Se hicieron pruebas de esfuerzo máximas limitadas por síntomas con determinación de consumo de oxígeno, ecocardiogramas en reposo y ventriculografías isotópicas en reposo y esfuerzo a los 2, 8 y 18 meses de evolución, y un tiempo medio de seguimiento clínico de 4,1 años. A todos se les prescribió un régimen de entrenamiento físico moderado o intenso, durante un año como mínimo. Se consideró disfunción sistólica severa cuando la fracción de eyección del ventrículo izquierdo fue menor de 35%. Resultados: Todos los parámetros ergométricos que expresaron capacidad funcional incrementaron significativamente en la evaluación del octavo mes (p< 0,0005), permaneciendo invariables a los 18. La fracción de eyección del ventrículo izquierdo media en reposo inicial fue de 28,3 ± 5,3%, la cual no mostró variaciones significativas con el esfuerzo ni con otros estudios evolutivos. La mortalidad total y la morbilidad de la serie fueron de 10,5% y 47,3%, respectivamente. Conclusión: El entrenamiento físico supervisado en pacientes infartados con disfunción sistólica severa de ventrículo izquierdo fue seguro y efectivo, y mejoró su calidad de vida, sin causar efectos negativos sobre la función ventricular.

Abstract Objective: To evaluate the feasibility and effects on the functional capacity of a supervised physical training programme carried out on patients with severe left ventricular systolic dysfunction after an acute myocardial infarction. Methods: The study included a total of 37 patients, males and females of any age, with a diagnosis of severe left ventricular systolic dysfunction after having suffered an acute myocardial infarction. They were consecutively included into the ambulatory programme of the Institute of Cardiology Rehabilitation Centre. Maximum effort tests, limited by symptoms, were performed to determine oxygen consumption. Echocardiograms were also performed at rest, with isotopic ventriculography at rest and then at 2, 8, and 18 months. The mean clinical follow-up was 4.1 years. They were all prescribed to a moderate or intense training programme for at least one year. Severe left ventricular systolic dysfunction was considered when the left ventricular ejection fraction was less than 35%. Results: All the ergometric parameters that expressed functional capacity increased significantly in the evaluation at 8 months (P< .0005), and remained at 18 months. The initial mean left ventricular ejection fraction at rest was 28.3 ± 5.3%, which showed no significant changes with effort or in the other evaluation times. The overall mortality and morbidity of the series was 10.5% and 47.3%, respectively. Conclusion: Supervised physical training in patients after an acute myocardial infarction and with severe left ventricular systolic dysfunction was safe and effective, and improved the quality of life, without causing negative effects on ventricular function.

The Activation of the Sox2 RR2 Pluripotency Transcriptional Reporter in Human Breast Cancer Cell Lines is Dynamic and Labels Cells with Higher Tumorigenic Potential.

The striking similarity displayed at the mechanistic level between tumorigenesis and the generation of induced pluripotent stem cells and the fact that genes and pathways relevant for embryonic development are reactivated during tumor progression highlights the link between pluripotency and cancer. Based on these observations, we tested whether it is possible to use a pluripotency-associated transcriptional reporter, whose activation is driven by the SRR2 enhancer from the Sox2 gene promoter (named S4+ reporter), to isolate cancer stem cells (CSCs) from breast cancer cell lines. The S4+ pluripotency transcriptional reporter allows the isolation of cells with enhanced tumorigenic potential and its activation was switched on and off in the cell lines studied, reflecting a plastic cellular process. Microarray analysis comparing the populations in which the reporter construct is active versus inactive showed that positive cells expressed higher mRNA levels of cytokines (IL-8, IL-6, TNF) and genes (such as ATF3, SNAI2, and KLF6) previously related with the CSC phenotype in breast cancer.

[Effects of long-term exercise training on left ventricular function and remodeling in patients with anterior wall myocardial infarction]. / Efectos del entrenamiento físico de larga duración sobre la función y remodelación del ventrículo izquierdo en pacientes con infarto miocárdico de pared anterior.

OBJECTIVE: To assess the effects of long-term exercise training on the function and remodeling of the left ventricle after myocardial infarction. METHODS: We studied 90 patients with a first acute anterior-wall myocardial infarction, all received conventional medical treatment. Symptom-limited maximal exercise stress tests, echocardiograms and effort-rest isotopic ventriculographies at 2, 6 and 12 months after myocardial infarction were performed; the follow-up time averaged 36.3±17 months. All patients joined a cardiac rehabilitation program with moderate or intense exercise training lasting at least a year. Of all patients, 41.1% suffered severe left ventricle dysfunction. RESULTS: Ergometric parameters that expressed functional capacity increased significantly (P<.0005) at the sixth month evaluation and remained unchanged after a year. There was significant decrease (P<.01) of exercise myocardial ischemia at 6 months. The variables that measured size and function of left ventricle did not change during evolution. Morbidity amounted to 16.7% and total mortality of the series was 13.3%, with 8.9% of cardiovascular cause. CONCLUSIONS: Long-term exercise training showed no deleterious effects on left ventricle function or remodeling and beneficial functional and clinical effects were obtained in these rehabilitated postinfarction patients.

Efectos del entrenamiento físico de larga duración sobre la función y remodelación del ventrículo izquierdo en pacientes con infarto miocárdico de pared anterior / Effects of long-term exercise training on left ventricular function and remodeling in patients with anterior wall myocardial infarction

Objetivo: Determinar los efectos del entrenamiento físico prolongado sobre la función y remodelación del ventrículo izquierdo después del infarto miocárdico. Métodos: Fueron estudiados 90 pacientes con un primer infarto miocárdico agudo, de localización anterior; todos recibieron el tratamiento médico convencional. Se realizaron pruebas de esfuerzo máximas, ecocardiogramas y ventriculografías isotópicas en reposo y esfuerzo a los 2, 6 y 12 meses de evolución; el seguimiento clínico medio fue de 36.3 ± 17 meses. A todos se les incorporó a un programa de rehabilitación cardíaca con entrenamiento físico moderado o intenso, durante un año como mínimo. Un 41.1% tuvieron una disfunción severa del ventrículo izquierdo. Resultados: Todos los parámetros ergométricos que expresaron capacidad funcional incrementaron significativamente en la evaluación del sexto mes (p < 0.0005), permaneciendo invariables al año. Se comprobó una disminución significativa (p < 0.01) de isquemia miocárdica al esfuerzo a los 6 meses. Las variables que midieron tamaño y función del ventrículo izquierdo no se modificaron evolutivamente. La mortalidad total de la serie fue un 13.3%, de causa cardiovascular un 8.9% y la morbilidad un 16.7%. Conclusiones: El entrenamiento físico prolongado no mostró efectos deletéreos sobre la función y remodelación del ventrículo izquierdo y se obtuvieron efectos funcionales y clínicos beneficiosos en estos pacientes infartados rehabilitados.

Objective: To assess the effects of long-term exercise training on the function and remodeling of the left ventricle after myocardial infarction. Methods: We studied 90 patients with a first acute anterior-wall myocardial infarction, all received conventional medical treatment. Symptom-limited maximal exercise stress tests, echocardiograms and effort-rest isotopic ventriculographies at 2, 6 and 12 months after myocardial infarction were performed; the follow-up time averaged 36.3 ± 17 months. All patients joined a cardiac rehabilitation program with moderate or intense exercise training lasting at least a year. Of all patients, 41.1% suffered severe left ventricle dysfunction. Results: Ergometric parameters that expressed functional capacity increased significantly (P< .0005) at the sixth month evaluation and remained unchanged after a year. There was significant decrease (P< .01) of exercise myocardial ischemia at 6 months. The variables that measured size and function of left ventricle did not change during evolution. Morbidity amounted to 16.7% and total mortality of the series was 13.3%, with 8.9% of cardiovascular cause. Conclusions: Long-term exercise training showed no deleterious effects on left ventricle function or remodeling and beneficial functional and clinical effects were obtained in these rehabilitated postinfarction patients.