Integración de la información experimental, expresión y ontologías para identificar efectos específicos inducidos por variantes glicosiladas de FSHrh

LORETI, NAZARETH; FRESNO, CRISTOBAL; FERNÁNDEZ, ELMER ANDRÉS; CAMPOS, STELLA

Cordoba

Congreso; 2do Congreso Argentino de Bioinformatica y Biologia Computacional; 2011

A2B2C (www.a2b2c.org.ar)

Background: Previous studies [1-3] have shown that different physiological conditions in women(menstrual cycle, full lactation, perimenopause and postmenopause) are associated with variations inthe degree of syalilation and complexity of FSH oligosaccharides. FSH carbohydrate moiety determinesnot only the hormone half-life but also the biological response in the target cell [4]. The objective of thepresent study was to determine the possible differential regulation of granulosa cell gene expresión byrecombinant human FSH (rhFSH) glycosylation variants.Materials and Methods: Four rhFSH preparations: more acidic, AC; less acidic, BA; those bearingcomplex, highly branched (UB) and hybrid type (FR) oligosaccharides were used to stimulate a humanovarian steroidogenic granulosa cell line (KGN). Cells were cultured with rhFSH glycosylation variants(20 ng/mL) for 24h. Purified total RNA was hybridized on Affimetrix GeneChip® Human Gene 1.0 STand Gene Expression and Ontology analysis were performed. A method to incorporate expression andexperimental information was designed in order to obtain biologically relevant information.Results: The above mentioned strategy allowed us to find cellular homeostasis, nucleosomeorganization, steroid binding, steroid dehydrogenase and ion transport enriched by AC. When BA wasadded, enriched Gene Ontology terms were mainly associated to important aspect of the reproductiveprocess such as gamete generation, cell differentiation (i.e.growth factors) and cell secretion.Programmed cell death as well as cell adhesion and peptide binding were linked to UB. Meanwhile,important aspects of cell endocrine activity such as steroid metabolism, ovarian follicle development,cytoskeleton (i.e. keratin), response to steroid hormones, cytokine receptors were identified in FBcultures.Conclusions: Integration of Gene expression, Experimental Design and Ontologies allowed us to easilyidentify meaningful biological information. In this case, results led us to recognize differentially regulatedgenes by rhFSH glycosylation variants. Moreover, it was possible to associate biological process andmolecular functions with specific characteristics of FSH carbohydrate moiety.