RESUMO
OBJECTIVES: Recombinant human interferon-beta (IFN-b) is a well-established treatment for multiple sclerosis (MS). The regulatory process for marketing authorization of biosimilars is currently under debate in certain countries. In the EU, EMEA has clearly defined the process including overarching and product-specific guidelines, which includes clinical testing. Biosimilarity needs to be based on comparability criteria, including at least molecular characterization, biological activity relevant for the therapeutic effect and relative bioavailability ("bioequivalence"). In the case of such complex diseases as MS, where the effect of treatment is not so directly measurable, in vitro tools can provide additional data to support comparability. Genomic microarrays assays might be useful to compare multisource biopharmaceuticals. The aim of the present study was to compare the pharmacodynamic genomic effects (in terms of transcriptional regulation) of two recombinant human IFN-I(2)1a preparations on lymphocytes of multiple sclerosis patients using a whole genome microarray assay. METHODS: We performed an ex vivo whole genome expression profiling of the effect of two preparations of IFN-I(2)1a on non-adherent mononuclears from five relapsing-remitting MS patients analyzing microarrays (CodeLink Human Whole Genome). Patients blood was drawn, PBMCs isolated and cultured in three different conditions: culture medium (control), 1,000 U/ml of IFN-I(2)1a (BLA- (STOFERON, Bio Sidus) and 1,000 U/ml of IFN-I(2)1a (REBIF, Serono) RNA was purified from non-adherent cells (mostly lymphocytes), amplified and hybridized. Raw data were generated by CodeLink proprietary software. Data normalization, quality control and analysis of differential gene expression between treatments were done using linear model for microarray data. Functional annotation analysis of IFN-I(2)1a MS treatment transcription was done using DAVID. RESULTS: Out of the approximately 45,000 human sequences examined, no evidence of differential regulation was found when both treatments were compared (minimum adjusted p-value > 0.999). The IFN-I(2)1a effect differentially regulated the expression of 868 genes. The expression of standard markers such as GTP cyclohidrolase, MxA, and OAS isoenzymes A and B changed as a consequence of the action of IFN-I(2)1a. CONCLUSIONS: This exhaustive and highly sensitive assay did not show differences in the genomic expression profile of these two products under the assayed experimental conditions. These results suggest that this technology might be useful for the initial comparison of biosimilars, being part of a comprehensive comparability program that includes clinical testing.
Assuntos
Perfilação da Expressão Gênica , Interferon beta/farmacologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Transcrição Gênica/efeitos dos fármacos , Análise por Conglomerados , Biologia Computacional/métodos , Composição de Medicamentos/métodos , Feminino , Genoma Humano , Humanos , Interferon beta-1a , Interferon beta/genética , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Masculino , Proteínas Recombinantes/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
El fondo de ojo fue estudiado en 50 ninos recien nacidos normales de termino, entre el 1o e 4o dia de vida, con el fin de describir las caracteristicas que lo diferencian, tanto del fondo de ojo del prematuro como del nino de mas de 1 semana de edad. Se estudio mediante oftamoscopia directa y control fotografico. Se utilizo fenilefrina al 10% con homatropina al 1% como colirio midriatico media hora antes del examen. Se realizo en habitacon en penumbra y sobre una camilla alta que permite al observador ubicarse, de pie, detras del nino. Un chupete provoca una relajacion muscular generalizada. Se clasificaron los hallazgos segun el tejido. En retina se encuentra una coloracion amarillenta grisacea en el polo posterior que va tornandose azulada hacia la periferia. Las arteriolas estan disminuidas en su calibre y la macula esta representada por una zona mas oscura rodeada de un halo claro. Entre un 3 y un 38%, segun diversos autores, se escuentran hemorragias retinales que se han constatado y que se reabsorben espontaneamente en pocos dias.
Assuntos
Fundo de Olho , Recém-Nascido , OftalmoscopiaRESUMO
El fondo de ojo fue estudiado en 50 ninos recien nacidos normales de termino, entre el 1o e 4o dia de vida, con el fin de describir las caracteristicas que lo diferencian, tanto del fondo de ojo del prematuro como del nino de mas de 1 semana de edad. Se estudio mediante oftamoscopia directa y control fotografico. Se utilizo fenilefrina al 10% con homatropina al 1% como colirio midriatico media hora antes del examen. Se realizo en habitacon en penumbra y sobre una camilla alta que permite al observador ubicarse, de pie, detras del nino. Un chupete provoca una relajacion muscular generalizada. Se clasificaron los hallazgos segun el tejido. En retina se encuentra una coloracion amarillenta grisacea en el polo posterior que va tornandose azulada hacia la periferia. Las arteriolas estan disminuidas en su calibre y la macula esta representada por una zona mas oscura rodeada de un halo claro. Entre un 3 y un 38%, segun diversos autores, se escuentran hemorragias retinales que se han constatado y que se reabsorben espontaneamente en pocos dias.
