Integrating system biology and intratumor gene therapy by trans-complementing the appropriate co-stimulatory molecule as payload in oncolytic herpes virus.

Systems biology has been applied at the multi-scale level within the cancer field, improving cancer prevention, diagnosis and enabling precision medicine approaches. While systems biology can expand the knowledge and skills for oncological treatment, it also represents a challenging expedition due to cancer complexity, heterogeneity and diversity not only between different cancer indications, but also in its evolution process through space and time. Here, by characterizing the transcriptional perturbations of the tumor microenvironment induced by oncolytic, we aimed to rationally design a novel armed oncolytic herpes virus. We found that intratumor oncovirotherapy with HSV-1 induces T-cell activation signatures and transcriptionally activates several costimulatory molecules. We identified differentially expressed costimulatory receptors and binding partners, where inducible co-stimulators (ICOS) resulted in the potentially most beneficial targeted therapy. Through an ex-vivo transcriptomic analysis, we explored the potential of arming an oncolytic virus as a combination therapy strategy; in particular, we engineered a targeted herpes virus encoding ICOSL (THV_ICOSL), which resulted in a significant improvement in tumor size control compared to unarmed parental virus. Also, combination with a PD-1 inhibitor enhanced antitumor efficacy as predictable by upregulation of PD-1 and ligands pair (PD-L1/PD-L2) upon oncolytic virus injection. Generation of the human version of this virus encoding hICOSL orthologue effectively and specifically activated human T cells by triggering the ICOS pathway. Our data support the data-driven generation of armed oncolytic viruses as combination immunotherapeutic with checkpoint inhibitors.

Glatiramer Acetate modulates ion channels expression and calcium homeostasis in B cell of patients with relapsing-remitting multiple sclerosis.

To investigate the effects of Glatiramer Acetate (GA) on B cells by an integrated computational and experimental approach. GA is an immunomodulatory drug approved for the treatment of multiple sclerosis (MS). GA effect on B cells is yet to be fully elucidated. We compared transcriptional profiles of B cells from treatment-naïve relapsing remitting MS patients, treated or not with GA for 6 hours in vitro, and of B cells before and after six months of GA administration in vivo. Microarrays were analyzed with two different computational approaches, one for functional analysis of pathways (Gene Set Enrichment Analysis) and one for the identification of new drug targets (Mode-of-action by Network Analysis). GA modulates the expression of genes involved in immune response and apoptosis. A differential expression of genes encoding ion channels, mostly regulating Ca2+ homeostasis in endoplasmic reticulum (ER) was also observed. Microfluorimetric analysis confirmed this finding, showing a specific GA effect on ER Ca2+ concentration. Our findings unveils a GA regulatory effect on the immune response by influencing B cell phenotype and function. In particular, our results highlight a new functional role for GA in modulating Ca2+ homeostasis in these cells.

VIH/SIDA: comportamiento epidemiológico de la transmisión vertical en el contexto general de la infección en Chile / HIV/AIDS: epidemiological behavior of vertical transmission in the general context of infection in Chile

ANTECEDENTES: En embarazadas seropositivas sin profilaxis antirretroviral la transmisión vertical (TV) del VIH es de 30%, cifra que disminuye bajo al 2% con un manejo integral de prevención. OBJETIVO: Conocer el comportamiento epidemiológico de la TV en Chile desde la creación del programa nacional de prevención. MÉTODO: Datos aportados por el Departamento de Epidemiología del MINSAL y CONASIDA (1984-2006). Estudió de situación VIH/SIDA por año, regiones, grupo etario y sexo, con especial énfasis en TV. RESULTADOS: Se notificaron 9.317 casos de VIH y 7.886 casos de SIDA (1984-2006), prevalencia en aumento hasta el 2003 con tendencia descendente posteriormente. La razón actual de VIH entre hombres y mujeres es de 4:1, con clara tendencia al aumento de notificación en mujeres. La principal vía de exposición sigue siendo la vía sexual (93,4%). El grupo más afectado está entre los 20-39 años (73,1% para VIH y 63,1% para SIDA), cabe destacar que entre 0-9 años se encuentra el 1,1% de los afectados por VIH y el 1,4% por SIDA, todos infectados por TV. CONCLUSIONES: La transmisión vertical de VIH en Chile es responsable de un bajo porcentaje de las personas notificadas de VIH/SIDA, pero es la causa de la totalidad de los niños afectados por la infección en nuestro país. Se observa una importante reducción de la TV, llegando a niveles muy cercanos a los objetivos ministeriales.

BACKGROUND: The risk of transmitting HIV from mother to unborn child, without any antiretroviral prophylaxis, reaches 30%. It can be reduced to less than 2% by implementing integral preventive strategies. OBJECTIVE: To assess the epidemiological profile of HIV vertical transmission in Chile since the implementation of the national AIDS transmission prevention program. METHOD: Data from the Epidemiology Department of the Health Ministry and the National Commission for AIDS grouped by year, location, ageandgender. Withspecial emphasis on vertical transmission. RESULTS: 9317 cases of HIV infection and 7886 cases of AIDS have been reported in Chile (1984-2006), with a decreasing tendency since 2003. Sexual exposure continues to be the primary route with 93.4% of the cases. The male/female ratio for case notif¡cation is 4:1, with a rising tendency among women. The most affected is between 20-39 years old (73% HIV notification and 63.1% for AIDS). Among children between 0-9 years of age is 1.1% of the HIV and 1.4% of AIDS notifications, all of them for vertical transmission. CONCLUSIONS: HIV vertical transmission in Chile is responsible for a small percentage of HIV/AIDS notifications, but it accounts for all of the infected children. There's been an important reduction in this transmission route, reaching prevalence close to the aimed rates, which can explained by the expanding prevention programs of the health policies implemented by the Health Ministry.

Hepatitis aguda viral durante el embarazo / Acute viral hepatitis during pregnancy

Acute hepatitis has a very low incidence disease during pregnancy. However, it may be an important cause of jaundice during gestation which in cases of viral etiology can have a very high morbidity and mortality risk to the mother and the fetus. The purpose of this review is to update the available knowledge regarding viral hepatitis during pregnancy including description of the main etiologies, transmission route, maternal-fetal risk and possible management.

La hepatitis aguda es una enfermedad de baja incidencia durante el embarazo; sin embargo, es una causa importante de ictericia durante el desarrollo de éste y en algunos casos presenta un alto riesgo de morbi-mortalidad materno-fetal, siendo la etiología principalmente viral. El propósito de este artículo es actualizar los conocimientos disponibles en la literatura médica respecto a hepatitis viral durante el embarazo, conocer cuáles son los agentes más prevalentes, vía de transmisión, riesgo para el binomio madre- hijo y eventual manejo.

[Acute viral hepatitis during pregnancy]. / Hepatitis aguda viral durante el embarazo.

Acute hepatitis has a very low incidence disease during pregnancy. However, it may be an important cause of jaundice during gestation which in cases of viral etiology can have a very high morbidity and mortality risk to the mother and the fetus. The purpose of this review is to update the available knowledge regarding viral hepatitis during pregnancy including description of the main etiologies, transmission route, maternal-fetal risk and possible management.

[Effectiveness of oral hypoglycemic drugs in the metabolic control of patients with gestational diabetes]. / Eficacia de los hipoglicemiantes orales en el control metabólico de pacientes con diabetes mellitus gestacional.

Gestational Diabetes is characterized by different degrees of glucose intolerance that produce a series of fetal and perinatal alterations. During many years, in those cases of gestational diabetes that did not respond to nutritional interventions, the use of insulin was a proven treatment to achieve metabolic control and thus a better perinatal outcome. At present, some new oral hypoglycemic drugs, from the family of sulfonylureas and biguanides, have been shown to be safe, of low cost, and apparently effective in the metabolic control of this disease. We review the publications that propose the use of oral hypoglycemic drugs for the metabolic control of gestational diabetes that does not respond to nutritional measures.

Eficacia de los hipoglicemiantes orales en el control metabólico de pacientes con diabetes mellitus gestacional / Effectiveness of oral hypoglycemic drugs in the metabolic control of patients with gestational diabetes

Gestational Diabetes is characterized by different degrees of glucose intolerance that produce a series of fetal and perinatal alterations. During many years, in those cases of gestational diabetes that did not respond to nutritional interventions, the use of insulin was a proven treatment to achieve metabolic control and thus a better perinatal outcome. At present, some new oral hypoglycemic drugs, from the family of sulfonylureas and biguanides, have been shown to be safe, of low cost, and apparently effective in the metabolic control of this disease. We review the publications that propose the use of oral hypoglycemic drugs for the metabolic control of gestational diabetes that does not respond to nutritional measures.

Síndrome de rubéola congénita: primer caso en Chile post programa de revacunación / Congenital rubella syndrome after rubella revaccination program: first chilean case

Se presenta el primer caso clínico de síndrome de rubéola congénita en Chile, posterior al programa de revacunación en 1999. Se describe el caso índice y se realiza una breve revisión actualizada del tema.

We present the first clinical case of congenital rubella syndrome (CRS) in Chile after the preconceptional vaccination campaign in 1999. This report describes the case and we present a brief review of CRS.

Induction of complete regressions of oncogene-induced breast tumors in mice.

Over the past decade, mouse models of cancer have come to resemble human disease much more closely than simple subcutaneous or orthotopic systems. Intervention strategies that work on these new model systems are more likely to have an impact clinically. We have shown recently that antiangiogenic stress imposed by loss of Id protein in endothelial progenitor cells results in dramatic central necrosis in breast tumors initiated in mice by overexpression of the her2/neu oncogene. Tumor cells remain viable at the periphery, perhaps via the hypoxic response pathway which allows the lesions to expand. Inhibition of this pathway by the inactivation of the Hif-1alpha chaperone Hsp90 in combination with antiangiogenic stress leads to the first reported complete regression of these aggressive breast tumors.

Divulsión del polo inferior: método seguro y eficaz para disminuir los partos espontáneos después de las 41 semanas / Membrane stripping: efficient and sure method to reduce the spontaneous labor after the 41 weeks

Objetivo: Conocer si la divulsión del polo inferior, realizada semanalmente desde las 38 semanas de gestación en pacientes de bajo riesgo, es un método seguro y eficaz para disminuir la incidencia de partos mayores de 41 semanas. Método: 110 embarazadas de bajo riesgo y de edad gestacional segura fueron randomizadas a divulsión de polo (casos) y no divulsión (controles). Desde las 38 semanas, tanto a las controles como las intervenidas, se les realizó tactos vaginales para evaluar pelvimetría, puntaje de Bishop y divulsión de polo en aquellas gestantes aleatorizadas y que cumplían las condiciones para poder realizar el procedimiento. El objetivo primario fue determinar la incidencia de parto espontáneo antes de las 41 semanas y el secundario fue evaluar la morbimortalidad materna y neonatal. Los datos fueron analizados por Chi cuadrado, test exacto de Fisher, test t de Student y test Wilcoxson rank-sum según correspondiera. Resultados: La población estudiada en ambos grupos fueron demográficamente similares. Las gestantes divulsionadas tuvieron significativamente (p<0,005) menor cantidad de partos (22,2 por ciento) después de las 41 semanas comparadas con el grupo de control (10,0 por ciento). Las complicaciones materno-neonales fueron similares en ambos grupos. Conclusiones: La divulsión de polo es un método seguro y eficaz en disminuir la incidencia de partos después de las 41 semanas.

Displasia esquelética letal: experiencia de nueve años y aproximación diagnóstica actual / Lethal skeletal dysplasia: experience of nine years and present diagnostic approach

Se presenta la experiencia de nueve años en el diagnóstico de las displasias esqueléticas letales mediante la descripción de seis casos clínicos. Se realiza una revisión actualizada del diagnóstico de estas patologías en el período antenatal.

Prevalencia de colonizacion por streptococcus agalactiae (grupo b) en el tercer trimestre del embarazo pesquisado en medio de cultivo no selectivo

El Streptococcus agalactiae es el principal agente bacteriano responsable de sepsis neonatal. Para evitar la infección perinatal se recomienda la pesquisa de la bacteria en región vagino-perianal durante el tercer trimestre, indicando tratamiento antibiótico durante el parto en aquellas gestantes colonizadas. La prevalencia de colonización varía según el tipo de cultivo utilizado (selectivo vs no selectivo). El objetivo de este estudio es conocer la prevalencia de colonización del Streptococcus agalactiae en la población de embarazadas controladas en nuestra maternidad durante los años 2001-2002. Se tomó cultivo en medio no selectivo, entre las 35-37 semanas de gestación a 1658 pacientes, encontrando una prevalencia de colonización vagino-perianal de 6,2%. Esta cifra es bastante menor a la comunicada en otros estudios en embarazadas chilenas, por lo que creemos importante la implementación de medios de cultivos selectivos para mejorar el rendimiento de pesquisa.

Schistosomiasis mansoni in low transmission areas: abdominal ultrasound.

In endemic areas with low prevalence and low intensity of infection, the diagnosis of hepatic pathology due to the Schistosoma mansoni infection is very difficult. In order to establish the hepatic morbidity, a double-blind study was achieved in Venezuelan endemic areas, with one group of patients with schistosomiasis and the other one of non-infected people, that were evaluated clinically and by abdominal ultrasound using the Cairo classification. Schistosomiasis diagnosis was established based on parasitologic and serological tests. The increase of the hepatic size at midclavicular and midsternal lines (in hepatometry) and the hard liver consistency were the clinical parameters able to differentiate infected persons from non infected ones, as well as the presence of left lobe hepatomegaly detected by abdominal ultrasound. The periportal thickening, especially the mild form, was frequent in all age groups in both infected and uninfected patients. There was not correlation between the intensity of infection and ultrasound under the current circumstances. Our data suggest that in Venezuela, a low endemic area of transmission of schistosomiasis, the hepatic morbidity is mild and uncommon. The Cairo classification seems to overestimate the prevalence of periportal pathology. The specificity of the method must be improved, especially for the recognition of precocious pathology. Other causes of hepatopathies must be investigated.

Schistosomiasis mansoni in low transmission areas. Abdominal ultrasound

In endemic areas with low prevalence and low intensity of infection, the diagnosis of hepatic pathology due to the Schistosoma mansoni infection is very difficult. In order to establish the hepatic morbidity, a double-blind study was achieved in Venezuelan endemic areas, with one group of patients with schistosomiasis and the other one of non-infected people, that were evaluated clinically and by abdominal ultrasound using the Cairo classification. Schistosomiasis diagnosis was established based on parasitologic and serological tests. The increase of the hepatic size at midclavicular and midsternal lines (in hepatometry) and the hard liver consistency were the clinical parameters able to differentiate infected persons from non infected ones, as well as the presence of left lobe hepatomegaly detected by abdominal ultrasound. The periportal thickening, especially the mild form, was frequent in all age groups in both infected and uninfected patients. There was not correlation between the intensity of infection and ultrasound under the current circumstances. Our data suggest that in Venezuela, a low endemic area of transmission of schistosomiasis, the hepatic morbidity is mild and uncommon. The Cairo classification seems to overestimate the prevalence of periportal pathology. The specificity of the method must be improved, especially for the recognition of precocious pathology. Other causes of hepatopathies must be investigated

The beta-amyloid precursor protein functions as a cytosolic anchoring site that prevents Fe65 nuclear translocation.

In this study we addressed the question of the intracellular localization of Fe65, an adaptor protein interacting with the beta-amyloid precursor protein (APP) and with the transcription factor CP2/LSF/LBP1. By using tagged Fe65 expression vectors, we observed that a significant fraction of Fe65 is localized in the nucleus of transfected COS7 cells. Furthermore, the isolation of nuclei from untransfected PC12 cells allowed us to observe that a part of the endogenous Fe65 is present in the nuclear extract. The analysis of Fe65 mutant constructs demonstrated that the region of the protein required for its nuclear translocation includes the WW domain, and that, on the other hand, a small fragment of 100 residues, including this WW domain, contains enough structural information to target a reporter protein (green fluorescent protein (GFP)-GFP) to the nucleus. To evaluate whether the Fe65-APP interaction could affect Fe65 intracellular trafficking, COS7 cells were cotransfected with APP(695) or APP(751) and with GFP-Fe65 expression vectors. These experiments demonstrated that Fe65 is no longer translocated to the nucleus when the cells overexpress APP, whereas the nuclear targeting of GFP-Fe65 mutants, unable to interact with APP, is unaffected by the coexpression of APP, thus suggesting that the interaction with APP anchors Fe65 in the cytosol.

Fe65 and the protein network centered around the cytosolic domain of the Alzheimer's beta-amyloid precursor protein.

A distinctive tract of all the forms of Alzheimer's disease is the extracellular deposition of a 40-42/43 amino acid-long peptide derived from the so-called beta-amyloid precursor protein (APP). This is a membrane protein of unknown function, whose short cytosolic domain has been recently demonstrated to interact with several proteins. One of these proteins, named Fe65, has the characteristics of an adaptor protein; in fact, it possesses three protein-protein interaction domains: a WW domain and two PID/PTB domains. The interaction with APP requires the most C-terminal PID/PTB domain, whereas the WW domain is responsible for the interaction with various proteins, one of which was demonstrated to be the mammalian homolog of the Drosophila enabled protein (Mena), which in turn interacts with the cytoskeleton. The second PID/PTB domain of Fe65 binds to the CP2/LSF/LBP1 protein, which is an already known transcription factor. The other proteins interacting with the cytosolic domain of APP are the G(o) heterotrimeric protein, APP-BP1 and X11. The latter interacts with APP through a PID/PTB domain and possesses two other protein-protein interaction domains. The small size of the APP cytodomain and the overlapping of its regions involved in the binding of Fe65 and X11 suggest the existence of competitive mechanisms regulating the binding of the various ligands to this cytosolic domain. In this short review the possible functional roles of this complex protein network and its involvement in the generation of Alzheimer's phenotype are discussed.

The Fe65 adaptor protein interacts through its PID1 domain with the transcription factor CP2/LSF/LBP1.

The neural protein Fe65 possesses three putative protein-protein interaction domains: one WW domain and two phosphotyrosine interaction/phosphotyrosine binding domains (PID1 and PID2); the most C-terminal of these domains (PID2) interacts in vivo with the Alzheimer's beta-amyloid precursor protein, whereas the WW domain binds to Mena, the mammalian homolog of Drosophila-enabled protein. By the interaction trap procedure, we isolated a cDNA clone encoding a possible ligand of the N-terminal PID/PTB domain of Fe65 (PID1). Sequence analysis of this clone revealed that this ligand corresponded to the previously identified transcription factor CP2/LSF/LBP1. Co-immunoprecipitation experiments demonstrated that the interaction between Fe65 and CP2/LSF/LBP1 also takes place in vivo between the native molecules. The localization of both proteins was studied using fractionated cellular extracts. These experiments demonstrated that the various isoforms of CP2/LSF/LBP1 are differently distributed among subcellular fractions. At least one isoform, derived from alternative splicing (LSF-ID), is present outside the nucleus; Fe65 was found in both fractions. Furthermore, transfection experiments with an HA-tagged CP2/LSF/LBP1 cDNA demonstrated that Fe65 interacts also with the nuclear form of CP2/LSF/LBP1. Considering that the analysis of Fe65 distribution in fractionated cell extracts demonstrated that this protein is present both in nuclear and non-nuclear fractions, we examined the expression of Fe65 deletion mutants in the two fractions. This analysis allowed us to observe that a small region N-terminal to the WW domain is phosphorylated and is necessary for the presence of Fe65 in the nuclear fraction.

Interaction of the phosphotyrosine interaction/phosphotyrosine binding-related domains of Fe65 with wild-type and mutant Alzheimer's beta-amyloid precursor proteins.

The two tandem phosphotyrosine interaction/phosphotyrosine binding (PID/PTB) domains of the Fe65 protein interact with the intracellular region of the Alzheimer's beta-amyloid precursor protein (APP). This interaction, previously demonstrated in vitro and in the yeast two hybrid system, also takes place in vivo in mammalian cells, as demonstrated here by anti-Fe65 co-immunoprecipitation experiments. This interaction differs from that occurring between other PID/PTB domain-containing proteins, such as Shc and insulin receptor substrate 1, and activated growth factor receptors as follows: (i) the Fe65-APP interaction is phosphorylation-independent; (ii) the region of the APP intracellular domain involved in the binding is larger than that of the growth factor receptor necessary for the formation of the complex with Shc; and (iii) despite a significant similarity the carboxyl-terminal regions of PID/PTB of Fe65 and of Shc are not functionally interchangeable in terms of binding cognate ligands. A role for Fe65 in the pathogenesis of familial Alzheimer's disease is suggested by the finding that mutant APP, responsible for some cases of familial Alzheimer's disease, shows an altered in vivo interaction with Fe65.