Structure-Activity Studies of Novel Di-substituted [1,2,5]oxadiazolo [3,4-b]pyrazine Analogs Targeting the A-loop Regulatory Site of p38 MAP Kinase.

INTRODUCTION: In the quest for novel allosteric inhibitors of the p38 MAP kinase, we recently described the A-loop regulatory site, identified by means of molecular modeling studies together with the disclosure of a small molecule hit with a moderate inhibitory profile. Starting from this structure, we subsequently identified two additional hits with simpler molecular structures from an in silico screening study, using a substructure search in the SciFinder database. After corroboration of their inhibitory profile, analysis of their structures permitted to conclude about the suitability of the [1,2,5]oxadiazolo[3,4-b]pyrazine (furazano[ 3,4-b]pyrazine) scaffold for the development of potent A-loop regulatory site p38 MAP kinase inhibitors. Accordingly, we report the synthesis and pharmacological evaluation of a series of di-substituted analogs with a potent inhibitory profile of p38 MAP kinase, as shown by in vitro assays of their capability to inhibit IL-1ß secretion in human monocyte-derived macrophages. OBJECTIVE: To find small molecule potent inhibitors of the p38 MAP kinase A-loop regulatory site. METHODS: Starting from this structure, we subsequently identified two additional hits with simpler molecular structures from an in silico screening study, using a substructure search in the SciFinder database. After corroboration of their inhibitory profile, we carried out a hit-tolead optimization process guided by molecular modeling using a [1,2,5]oxadiazolo[3,4- b]pyrazine (furazano[3,4-b]pyrazine) scaffold. RESULTS: We report the synthesis and pharmacological evaluation of a series of di-substituted analogs with a potent inhibitory profile of p38 MAP kinase, as shown by in vitro assays of their capability to inhibit IL-1ß secretion in human monocyte-derived macrophages. CONCLUSION: We describe in the present work a series of [1,2,5]oxadiazolo[3,4-b]pyrazine (furazano[3,4-b]pyrazine), which are potent inhibitors of IL-1ß secretion in human monocytederived macrophages allosteric modulators of the p38 MAP kinase A-loop regulatory site.

Discovery of novel 2,3,5-trisubstituted pyridine analogs as potent inhibitors of IL-1ß via modulation of the p38 MAPK signaling pathway.

Interleukin-1ß is a central mediator of innate immune responses and inflammation. It plays a key role in a wide variety of pathologies, ranging from autoinflammatory diseases to metabolic syndrome and malignant tumors. It is well established that its inhibition results in a rapid and sustained reduction in disease severity, underlining the importance of having a repertoire of drugs of this class. At present, there are only three interleukin-1ß blockers approved in the clinic. All of them are biologics, requiring parenteral administration and resulting in expensive treatments. In an exercise to identify small molecule allosteric inhibitors of MAP kinases, we discovered a series of compounds that block IL-1ß release produced as a consequence of a stimulus involved in triggering an inflammatory response. The present study reports the hit-to-lead optimization process that permitted the identification of the compound 13b (AIK3-305) an orally available, potent and selective inhibitor of IL-1ß. Furthermore, the study also reports the results of an in vivo efficacy study of 13b in a LPS endotoxic shock model in male BALB/c mice, where IL-1ß inhibition is monitored in different tissues.

Mesenchymal stem cell's secretome promotes selective enrichment of cancer stem-like cells with specific cytogenetic profile.

Cancer stem cells (CSCs) are responsible for tumor initiation, metastasis and cancer recurrence, however the involvement of microenvironment is crucial. Here, we have analyzed how human mesenchymal stem cells (MSCs)-derived conditioned medium (CM) affect colon and melanoma CSCs enrichment and maintenance. Our results strongly suggest that the secretome of CM-MSCs selects and maintains subpopulations with high expression of CSCs markers and ALDH1 activity, low proliferation rates with G1 phase arrest, and notably retain in vivo these properties. Cytogenetic analyses indicated that CM-cultured cells contain alterations in chromosome 17 (17q25). Subsequent SKY-FISH analyses suggested that genes located in 17q25 might be involved in stem-cell maintenance. The characterization of secreted proteins present in CM-MSCs revealed that four cytokines and seven growth factors are directly linked to the CSCs enrichment reported in this study. Further analyses revealed that the combination of just IL6 and HGF is enough to provide cancer cells with better stemness properties. In conclusion, this study demonstrates how specific chromosomal alterations present in CSCs subpopulations might represent an advantage for their in vitro maintenance and in vivo stemness properties.

Protective Effects of Human and Mouse Soluble Scavenger-Like CD6 Lymphocyte Receptor in a Lethal Model of Polymicrobial Sepsis.

Sepsis still constitutes an unmet clinical need, which could benefit from novel adjunctive strategies to conventional antibiotic therapy. The soluble form of the scavenger-like human CD6 lymphocyte receptor (shCD6) binds to key pathogenic components from Gram-positive and -negative bacteria and shows time- and dose-dependent efficacy in mouse models of monobacterial sepsis. The objective of the present work was to demonstrate the effectiveness of infusing mouse and human sCD6 by different systemic routes, either alone or as adjunctive therapy to gold standard antibiotics, in a lethal model of polymicrobial sepsis. To this end, C57BL/6 mice undergoing high-grade septic shock induced by cecal ligation and puncture (CLP; ≥90% lethality) were infused via the intraperitoneal (i.p.) or intravenous (i.v.) route with shCD6 at different doses and time points, either alone or in combination with imipenem/cilastatin (I/C) at a dose of 33 mg/kg of body weight every 8 h. Significantly reduced mortality and proinflammatory cytokine levels were observed by i.p. infusion of a single shCD6 dose (1.25 mg/kg) 1 h pre- or post-CLP. When using the i.v. route, mice survival was significantly extended by starting shCD6 infusion at later time points post-CLP (up to 6 h after CLP). Significant adjunctive effects on mouse survival were observed by i.p. or i.v. infusion of shCD6 in combination with i.p. I/C post-CLP. Similar results were obtained in mice expressing high sustained levels (5 to 10 µg/ml) of mouse sCD6 in serum by means of transduction with hepatotropic adeno-associated virus (AAV). Taken together, the data support the conserved antibacterial effects of human and mouse sCD6 and their use as adjunctive therapy in experimental models of complex and severe polymicrobial sepsis.

Functionalized immunostimulating complexes with protein A via lipid vinyl sulfones to deliver cancer drugs to trastuzumab-resistant HER2-overexpressing breast cancer cells.

BACKGROUND: Around 20%-30% of breast cancers overexpress the proto-oncogene human epidermal growth receptor 2 (HER2), and they are characterized by being very invasive. Therefore, many current studies are focused on testing new therapies against tumors that overexpress this receptor. In particular, there exists major interest in new strategies to fight breast cancer resistant to trastuzumab (Tmab), a humanized antibody that binds specifically to HER2 interfering with its mitogenic signaling. Our team has previously developed immunostimulating complexes (ISCOMs) as nanocapsules functionalized with lipid vinyl sulfones, which can incorporate protein A and bind to G immunoglobulins that makes them very flexible nanocarriers. METHODS AND RESULTS: The aim of this in vitro study was to synthesize and evaluate a drug delivery system based on protein A-functionalized ISCOMs to target HER2-overexpressing cells. We describe the preparation of ISCOMs, the loading with the drugs doxorubicin and paclitaxel, the binding of ISCOMs to alkyl vinyl sulfone-protein A, the coupling of Tmab, and the evaluation in both HER2-overexpressing breast cancer cells (HCC1954) and non-overexpressing cells (MCF-7) by flow cytometry and fluorescence microscopy. Results show that the uptake is dependent on the level of overexpression of HER2, and the analysis of the cell viability reveals that targeted drugs are selective toward HCC1954, whereas MCF-7 cells remain unaffected. CONCLUSION: Protein A-functionalized ISCOMs are versatile carriers that can be coupled to antibodies that act as targeting agents to deliver drugs. When coupling to Tmab and loading with paclitaxel or doxorubicin, they become efficient vehicles for the selective delivery of the drug to Tmab-resistant HER2-overexpressing breast cancer cells. These nanoparticles may pave the way for the development of novel therapies for poor prognosis resistant patients.

Meroxest improves the prognosis of immunocompetent C57BL/6 mice with allografts of E0771 mouse breast tumor cells.

INTRODUCTION: Recently, we have reported the antitumor properties of a new family of synthetic merosesquiterpenes, among which meroxest is highlighted, since it has high activity and specificity for ER+ breast cancer cells. In this paper, we characterize allografts of ER+ E0771 mouse breast tumor cells in immunocompetent C57BL/6 mice, and also analyze the effect of meroxest on the prognosis of the disease. MATERIAL AND METHODS: Twenty female C57BL/6 mice were injected with 106 E0771 cells. Once the tumors reached the appropriate size, the mice were divided into two groups, one control and another treated orally with 15 mg/kg of meroxest. After 20 days, tumor samples were taken for histopathological study and for determination of the expression of the prognostic markers Ki67 and vascular endothelial growth factor (VEGF) by immunofluorescence. RESULTS: In sections stained with hematoxylin-eosin, we observed that tumors have a well-defined capsule enclosing E0771 tumor cells. The central area of tumors contains necrotic regions with leukocyte infiltration. Meroxest treatment significantly reduces tumor size (68%, p < 0.05), induces changes in its structure, decreases the degree of leukocyte infiltration, and significantly reduces the expression of Ki67 (33%, p < 0.05) and VEGF (82%, p < 0.05). CONCLUSIONS: Meroxest improves the prognosis of mice since it reduces leukocyte infiltration, and decreases the expression of Ki67 and VEGF markers. Consequently, the merosesquiterpene could become a useful antiangiogenic drug in the treatment of breast cancer. These results encourage us to deepen the study of meroxest, in order to find more evidence that supports the convenience of its evaluation in a clinical study or trial.

Antioxidant Intake and Antitumor Therapy: Toward Nutritional Recommendations for Optimal Results.

The role of the induction of oxidative stress as the mechanism of action of many antitumor drugs is acquiring an increasing interest. In such cases, the antitumor therapy success may be conditioned by the antioxidants present in our own body, which can be synthesized de novo (endogenous) or incorporated through the diet and nutritional supplements (exogenous). In this paper, we have reviewed different aspects of antioxidants, including their classification, natural sources, importance in diet, consumption of nutritional supplements, and the impact of antioxidants on health. Moreover, we have focused especially on the study of the interaction between antioxidants and antitumor therapy, considering both radiotherapy and chemotherapy. In this regard, we found that the convenience of administration of antioxidants during cancer treatment still remains a very controversial issue. In general terms, antioxidants could promote or suppress the effectiveness of antitumor treatment and even protect healthy tissues against damage induced by oxidative stress. The effects may depend on many factors discussed in the paper. These factors should be taken into consideration in order to achieve precise nutritional recommendations for patients. The evidence at the moment suggests that the supplementation or restriction of exogenous antioxidants during cancer treatment, as appropriate, could contribute to improving its efficiency.

Pattern Recognition by CD6: A Scavenger-Like Lymphocyte Receptor.

CD6, one of the first antigens to be identified on T cells, is a membrane glycoprotein that physically associates with the antigen receptor complex. Because of this, its main function seems to involve the modulation of TCR-mediated signaling pathways. However, growing evidence indicates that this ancient and conserved scavenger-like receptor may also play a role as pattern recognition receptor (PRR), similar to other members of the scavenger receptor cysteine rich superfamily (SRCR-SF). Here, we discuss the functional interactions of CD6 with microbe- and damage-associated signals and the potential use of soluble forms of CD6 in the therapeutic treatment of bacterial infections, in particular multi-drug resistant bacterial strains. Importantly, microbe recognition by CD6 may also have functional consequences on T cell activation and differentiation, which remain to be explored.

First enantiospecific syntheses of marine merosesquiterpenes neopetrosiquinones a and B: evaluation of biological activity.

The first enantiospecific syntheses of neopetrosiquinones A (6) and B (7), two merosesquiterpenes isolated from the deep-water sponge Neopetrosia cf. proxima, from the labdane diterpene trans-communic acid (10) have been achieved. A key step of the synthetic sequence is the simultaneous aromatization of the C ring and the benzylic oxidation on C-7 of an advanced intermediate, mediated by the oxygen-DDQ system. The in vitro antiproliferative activities of neopetrosiquinone B (7) and of the synthetic intermediates 8 and 9 against human breast (MCF-7), lung (A-549), and colon (T-84) tumor cell lines have been assayed. The most potent was compound 9 (IC50 = 4.1 µM), which was twice as active as natural compound 7 (IC50 = 8.3 µM) against A-549 cells. In addition, the treatment with these compounds resulted in an induction of apoptosis. These findings indicate that the terpene benzoquinones reported here might be potentially useful as anticancer agents.

Novel merosesquiterpene exerts a potent antitumor activity against breast cancer cells in vitro and in vivo.

This article describes the antitumor properties of a new family of merosesquiterpenes, which were synthesized by Diels-Alder cycloaddition of the labdane diene trans-communic acid, highly abundant in Cupressus sempervirens, or its methyl ester, with the appropriate dienophile. These compounds demonstrated potent cytotoxic activity in vitro against human breast, colon, and lung tumor cells. We highlight the elevated activity (IC50: 0.35 ± 0.10 µM) and specificity (TI: 9) of compound 13 against the MCF-7 line, which corresponds to the most prevalent breast cancer cell subtype, luminal A. It was found that compound 13 exerts its anti-tumor action by inducing oxidative stress, arresting the cell cycle in stages G0-G1, and activating apoptosis, which are all associated with low cyclin D1 regulation, pRb hypophosphorylation, increased expression of p27 and p53, and poly (ADP-ribose) polymerase (PARP) fractioning. Epithelial-mesenchymal transition, a phenomenon associated with metastasis promotion and a worsened prognosis also appeared to be inhibited by compound 13. In addition, it markedly reduced tumor development in immunocompetent C57BL/6 mice with allografts of E0771 mouse breast tumor cells (luminal A subtype). According to these findings, this new family of compounds, especially compound 13, may be highly useful in the treatment of human breast cancer.

Application of nanotechnology in the treatment and diagnosis of gastrointestinal cancers: review of recent patents.

Gastrointestinal cancers remain one of the main causes of death in developed countries. The main obstacles to combating these diseases are the limitations of current diagnostic techniques and the low stability, availability, and/or specificity of pharmacological treatment. In recent years, nanotechnology has revolutionized many fields of medicine, including oncology. The association of chemotherapeutic agents with nanoparticles offers improvement in the solubility and stability of antitumor agents, avoidance of drug degradation, and reductions in therapeutic dose and toxicity, increasing drug levels in tumor tissue and decreasing them in healthy tissue. The use of specific molecules that drive nanoparticles to the tumor tissue represents a major advance in therapeutic specificity. In addition, the use of nanotechnology in contrast agents has yielded improvements in the diagnosis and the follow-up of tumors. These nanotechnologies have all been applied in gastrointestinal cancer treatment, first in vitro, and subsequently in vivo, with promising results reported in some clinical trials. A large number of patents have been generated by nanotechnology research over recent years. The objective of this paper is to review patents on the clinical use of nanoparticles for gastrointestinal cancer diagnosis and therapy and to offer an overview of the impact of nanotechnology on the management of this disease.

Bozepinib, a novel small antitumor agent, induces PKR-mediated apoptosis and synergizes with IFNα triggering apoptosis, autophagy and senescence.

Bozepinib [(RS)-2,6-dichloro-9-[1-(p-nitrobenzenesulfonyl)-1,2,3,5-tetrahydro-4,1-benzoxazepin-3-yl]-9H-purine] is a potent antitumor compound that is able to induce apoptosis in breast cancer cells. In the present study, we show that bozepinib also has antitumor activity in colon cancer cells, showing 50% inhibitory concentration (IC50) values lower than those described for breast cancer cells and suggesting great potential of this synthetic drug in the treatment of cancer. We identified that the double-stranded RNA-dependent protein kinase (PKR) is a target of bozepinib, being upregulated and activated by the drug. However, p53 was not affected by bozepinib, and was not necessary for induction of apoptosis in either breast or colon cancer cells. In addition, the efficacy of bozepinib was improved when combined with the interferon-alpha (IFNα) cytokine, which enhanced bozepinib-induced apoptosis with involvement of protein kinase PKR. Moreover, we report here, for the first time, that in combined therapy, IFNα induces a clear process of autophagosome formation, and prior treatment with chloroquine, an autophagy inhibitor, is able to significantly reduce IFNα/bozepinib-induced cell death. Finally, we observed that a minor population of caspase 3-deficient MCF-7 cells persisted during long-term treatment with lower doses of bozepinib and the bozepinib/IFNα combination. Curiously, this population showed ß-galactosidase activity and a percentage of cells arrested in S phase, that was more evident in cells treated with the bozepinib/IFNα combination than in cells treated with bozepinib or IFNα alone. Considering the resistance of some cancer cells to conventional chemotherapy, combinations enhancing the diversity of the cell death outcome might succeed in delivering more effective and less toxic chemotherapy.

Cadmium modifies the cell cycle and apoptotic profiles of human breast cancer cells treated with 5-fluorouracil.

Industrialisation, the proximity of factories to cities, and human work activities have led to a disproportionate use of substances containing heavy metals, such as cadmium (Cd), which may have deleterious effects on human health. Carcinogenic effects of Cd and its relationship with breast cancer, among other tumours, have been reported. 5-Fluorouracil (5-FU) is a fluoropyrimidine anticancer drug used to treat solid tumours of the colon, breast, stomach, liver, and pancreas. The purpose of this work was to study the effects of Cd on cell cycle, apoptosis, and gene and protein expression in MCF-7 breast cancer cells treated with 5-FU. Cd altered the cell cycle profile, and its effects were greater when used either alone or in combination with 5-FU compared with 5-FU alone. Cd significantly suppressed apoptosis of MCF-7 cells pre-treated with 5-FU. Regarding gene and protein expression, bcl2 expression was mainly upregulated by all treatments involving Cd. The expression of caspase 8 and caspase 9 was decreased by most of the treatments and at all times evaluated. C-myc expression was increased by all treatments involving Cd, especially 5-FU plus Cd at the half time of treatment. Cd plus 5-FU decreased cyclin D1 and increased cyclin A1 expression. In conclusion, our results indicate that exposure to Cd blocks the anticancer effects of 5-FU in MCF-7 cells. These results could have important clinical implications in patients treated with 5-FU-based therapies and who are exposed to high levels of Cd.

The chemotherapeutic drug 5-fluorouracil promotes PKR-mediated apoptosis in a p53-independent manner in colon and breast cancer cells.

The chemotherapeutic drug 5-FU is widely used in the treatment of a range of cancers, but resistance to the drug remains a major clinical problem. Since defects in the mediators of apoptosis may account for chemo-resistance, the identification of new targets involved in 5-FU-induced apoptosis is of main clinical interest. We have identified the ds-RNA-dependent protein kinase (PKR) as a key molecular target of 5-FU involved in apoptosis induction in human colon and breast cancer cell lines. PKR distribution and activation, apoptosis induction and cytotoxic effects were analyzed during 5-FU and 5-FU/IFNα treatment in several colon and breast cancer cell lines with different p53 status. PKR protein was activated by 5-FU treatment in a p53-independent manner, inducing phosphorylation of the protein synthesis translation initiation factor eIF-2α and cell death by apoptosis. Furthermore, PKR interference promoted a decreased response to 5-FU treatment and those cells were not affected by the synergistic antitumor activity of 5-FU/IFNα combination. These results, taken together, provide evidence that PKR is a key molecular target of 5-FU with potential relevance in the clinical use of this drug.

Supervivencia de pacientes con cáncer de pulmón de célula no pequeña: en cuatro instituciones de Colombia / Superlife in patients with lung cancer no smal cells: in four Colombian institutes

Describir los resultados del tratamiento en pacientes con carcinoma de células no pequeñas avanzado. El 70 % de los pacientes debutan con enfermedad avanzada con supervivencia global de 5 años menor del 13 %. MÉTODOS: Se analizaron variables clínicas de 176 pacientes tratados en Bogotá. RESULTADOS: Promedio de edad 64 años, 64 % hombres 51 % tenía ECOG ≤1. El 90 % tenía compromiso metastásico principalmente a nivel pulmonar y en sistema nervioso central. El 76 % recibió terapia combinada como primera línea; 126 sujetos fueron tratados con (cisplatino 38/21,6 % y carboplatino 88 / 50% más algún agente de tercera generación o etopósido. Treinta y cuatro pacientes recibieron monoterapia, ocho, erlotinib. Se suministraron 3,7 ± 1,7 ciclos por paciente, la tasa de respuesta global fue 29 %, beneficio clínico 39 %, la mediana del tiempo de progresión 3,4 meses. La tercera parte de la población recibió segunda línea; en este grupo tasa de respuesta completa 15,8 %, beneficio clínico 33,8 % la mediana de tiempo libre de progresión 3,1 meses. El 69,3 % había muerto, la mediana de sobrevida global para la población total fue 9,2 meses, la proporción de sujetos vivos a 1 año 26 %. Variables como sexo, estado funcional, historia de tabaquismo, administración de segunda línea, adenocarcinoma, influyeron sobre la supervivencia. CONCLUSIONES: El comportamiento clínico y los objetivos del tratamiento de los pacientes incluidos en este estudio son similares a los reportados en la literatura médica.

Describing results of treatment in patients suffering advanced no small cell according to routine clinical practice. The 70 % of patients suffering debut with advanced disease are determining less than 13 % overall survival in 5 years. METHODS: 176 patients treated in Bogotá. RESULTS: The mean age was 64 years; 64 % were male 51 % had ≤1 ECOG. The 90 % had metastasis, mainly in the lung and in central nervous system. 76 % received combined therapy as the first line of intervention; 126 subjects were treated with a platinum compounds (cisplatin 38/21.6 % carboplatin 88/50 %) plus some third generation agent or etoposide. Thirty-four received monotherapy and eight erlotinib; 3.7 ± 1.7 cycles per patient were supplied on average, overall response rate was 29 %, clinical benefit 39 % and the median time to progression was 3.4 months. Only the third part of the population received a second line intervention; in this group overall response was 15.8 %, clinical benefit 33.8 % and free time progression was 3.1 months. When follow-up finished, 69.3 % had died, median the overall survival was 9.2 months 26 % of the subjects lived for up to one year. Some variables such gender, functional state, history of smoking, administering second line agent and a diagnosis of adenocarcinoma influenced in superlife. CONCLUSIONS: Clinical behaviour and outcomes treatment of patients included in this study were similar to those previously reported in the medical literature.

Mieloptisis. Viejos aspectos, nuevos conceptos / Myelophthisis. Old aspects, new concepts

Introducción: la mieloptisis es la infiltración de la médula ósea por células no hematopoyéticas.Objetivo: realizar un análisis detallado de 89 sujetos con mieloptisis tratados entre 1991 y 2006.Material y métodos: se revisaron las historias clínicas de 138 pacientes en quienes se encontró leucoeritroblastosis. De éstos, se seleccionaron 89 sujetos mayores de 16 años, con diagnóstico de alguna neoplasia sólida y mieloptisis.Resultados: la edad promedio fue 47,5 ± 17,2 años, la distribución por sexo fue homogénea y el 62 por ciento tuvo un IK ≥60 por ciento. Veintisiete pacientes (30 por ciento) tenían cáncer de mama, patología seguida por los tumores de primario desconocido (27 por ciento), el rabdomiosarcoma (10 por ciento), adenocarcinoma de próstata (10 por ciento), el carcinoma gástrico (7 por ciento) y otras neoplasias sólidas (22 por ciento). En el momento en que se documentó la mieloptisis, 72 por ciento y 50 por ciento tenían metástasis óseas y viscerales, respectivamente; 81 pacientes presentaron anemia (Hb 9,8 ± 1,2 gr/dl), la mediana del recuento plaquetario fue 174.000 y el promedio de leucocitos 24.283 ± 5.447. Cuarenta y tres pacientes recibieron quimioterapia después del diagnóstico de infiltración medular, tras lo que se evidenció normalidad en el recuento leucocitario en 40 por ciento de los casos. Se presentaron nueve episodios de neutropenia febril, la mediana de supervivencia global (SG) a partir del diagnóstico de la neoplasia fue 13,8 meses, y después de la mieloptisis, 2,2 meses. Los factores relacionados con una menor supervivencia fueron: la presencia de Hb ≤8,5 gr/dl (HR: 0,54, IC95 por ciento 0,32-0,95; p = 0.04), >3 sitios de metástasis (HR: 0,67, IC95 por ciento 0,45-0,92; p = 0.03), enfermedad visceral (HR: 0,72, IC95 por ciento 0,66-0,89; p = 0.04) y la neutropenia febril por quimioterapia (HR: 0,52, IC95 por ciento 0,37-0,60; p = 0.02).Conclusiones: la mieloptisis es una condición grave que modifica la SG de los pacientes con tumores sólidos. El tratamiento de este subgrupo debe seleccionarse teniendo en cuenta su toxicidad hematológica.

Supervivencia en pacientes con adenocarcinoma de pulmón metastásico y registro de las primeras mutaciones en el receptor para el factor de crecimiento epidérmico documentado en Colombia: Estudio del ONCOLGroup / Survival of advanced lung adenocarcinoma patients and report of the first epidermal growth factor receptor mutations recognized in Colombia: ONCOLGroup study

Introducción: la incidencia del adenocarcinoma de pulmón (ADCP) se ha incrementado en 100 por ciento desde 1950. Material y métodos: realizar una descripción detallada de los desenlaces de 147 pacientes con ADCP avanzado tratados en Bogotá entre los años 2000 y 2007. Se estimaron la respuesta global (TRG), el beneficio clínico (BC), el tiempo libre de progresión (TLP) y la supervivencia global (SG). En un subgrupo de pacientes se estudiaron las mutaciones en el gen EGFR. Resultados: el promedio de edad fue 66±12.8 años, 78 fueron mujeres, el 40 por ciento nunca había tenido exposición al humo por combustión del tabaco y en 119 casos se encontró un estado funcional = 70 por ciento. El sitio dominante de metástasis fue el sistema nervioso seguido por la afectación pulmonar. El 69 por ciento de los pacientes recibió terapia combinada con platino como primera línea de intervención y 32 sujetos (22 por ciento) recibieron erlotinib en algún momento del tratamiento de la enfermedad. La respuesta a la primera línea fue evaluable en 110 pacientes (74.8 por ciento); la TRG fue 28 por ciento, el BC fue 39 por ciento y la mediana del TLP fue de 3.7 meses (0.6-18.2). En 46 pacientes se administró una segunda línea con la que se obtuvo una TRG de 8 por ciento, un BC del 25 por ciento y un TLP de 3.7 meses (2.1-17). La mediana de SG para la población total fue 9.8 meses (6.3-19). Algunas variables como el ECOG, la ausencia de tabaquismo, la administración de una segunda o tercera línea de tratamiento y el haber recibido erlotinib, influyeron positivamente sobre este desenlace. En 10 casos seleccionados se realizó el perfil mutacional para el EGFR, cuatro presentaron alteraciones en el exón 19 (dos pacientes mostraron una inserción de 3 nucleótidos ricos en serina (L747_S750) que no había sido reportada previamente) y dos tuvieron la mutación del E21 (L858R). Para este subgrupo, la TRG al erlotinib fue 85 por ciento y en cinco casos la SG superó los 16 meses. Conclusiones: los desenlaces de los pacientes con ADCP avanzado en Colombia son similares a los descritos en otros países de América Latina. En nuestra serie se identificaron seis pacientes con mutaciones del EGFR.

BRCA1: a new genomic marker for non-small-cell lung cancer.

We are all aware of the recent rapid changes in cancer management mostly achieved with emerging new data regarding tumor biology. Currently, research in oncology is mainly focused on identifying the unique molecular characteristics of neoplasms and developing new targeted drugs to treat them. Although some tumors have specific genetic alterations that set off a cause-and-effect process after targeted treatment, those who work in the lung cancer field recognize that this is a more complex disease in which various genetic disorders carry its distinctive aggressiveness. At this time, the efforts of the scientific community are directed toward the identification of predictive markers to customize treatment based on specific genomic or protein expression profiles of individual tumors. This report provides a review on the breast cancer susceptibility gene 1, a promising gene determinant of response to different types of chemotherapy and its potential applications as a new molecular marker in lung cancer.

A sensitive method for detecting EGFR mutations in non-small cell lung cancer samples with few tumor cells.

BACKGROUND: Detection of epidermal growth factor receptor (EGFR) mutations in advanced non-small cell lung cancer (NSCLC) patients has relied on DNA purification from biopsies, amplification, and sequencing. However, the number of tumor cells in a sample is often insufficient for EGFR assessment. METHODS: We prospectively screened 1380 NSCLC patients for EGFR mutations but found that 268 were not evaluable because of insufficient tumor tissue. We therefore developed and validated a method of detecting EGFR mutations in these samples. Tumor cells were microdissected into polymerase chain reaction buffer and amplified. EGFR mutations were detected by length analysis of fluorescently labeled polymerase chain reaction products and TaqMan assay. RESULTS: We determined EGFR status in 217 (81%) of the 268 primary NSCLC samples not evaluable in our original study-fresh and paraffin-embedded with less than 150 cells. Exon 19 deletions were detected in 11.5% of patients and exon 21 L858R mutations in 5.5%. In addition, the exon 20 T790M mutation was detected in 6 of 15 (40%) patients at the time of progression to erlotinib. The primary, sensitive mutation was present in all tumor cells, whereas the T790M mutation was absent in some groups. CONCLUSIONS: The method presented here eliminates the need for DNA purification and allows for detection of EGFR mutations in samples containing as few as eight cancer cells.

Intravitreous hepatocyte growth factor in patients with proliferative diabetic retinopathy: a case-control study.

The aim of the present study was to evaluate the vitreous levels of hepatocyte growth factor (HGF) in patients with proliferative diabetic retinopathy (PDR) and to investigate its relationship with vascular endothelial growth factor (VEGF) and retinopathy activity. In addition, the relationship between intravitreous HGF levels and the presence of epiretinal membranes (ERM), as well as the expression of c-Met in ERM were also investigated. In this case-control study, serum and vitreous samples as well as ERM specimens were obtained during vitrectomy from 28 diabetic patients with PDR and 30 non-diabetic control subjects. HGF and VEGF were determined by ELISA and c-Met expression by immunohistochemistry. Vitreal levels of both VEGF and HGF were higher in patients with PDR in comparison with the control group (p<0.0001). However, after correcting for total vitreous protein concentration, HGF (ng/mg of proteins) was lower in diabetic patients than in non-diabetic control subjects (p=0.02). No correlation was detected between the vitreal levels of HGF and VEGF. In addition, intravitreous VEGF but not HGF was found to be related to PDR activity. Both diabetic patients and non-diabetic patients in whom ERM had been excised presented higher HGF intravitreous levels. Finally, a significant expression of c-Met in ERM membranes were observed in both diabetic patients with PDR and in non-diabetic subjects. In conclusion, both HGF and VEGF increased, but were not related, in the vitreous fluid of diabetic patients with PDR. Our findings suggest that HGF is related to pathological conditions in which fibroproliferative processes or wound healing are involved rather than with angiogenesis itself.