New analytical scenarios and new approaches in the embryonic genetic investigation of the macromolecular alterations responsible for the neurodegenerative diseases

Abstract

Systematic, genome-wide interrogations have identified hundreds of genes, including several transcription factors, which have expression patterns tightly correlated with ES cell differentiation. OCT4, SOX2 and NANOG constitute a triad of transcription factors, identified as crucial for the maintenance of ES cells self-renewal and pluripotent state.The principal aim of our project was to induce the differentiation process of embryo-derived stem cells into neural cells (neurons, glial cells), to follow during the differentiation process the changing in the expression of characteristic stemness markers (OCT4, SOX2 and NANOG) responsible for the regulatory networks involved in embryo-derived stem cells pluripotency, whose understanding is fundamental for any potential therapeutic application. For this reason the use of advanced spectroscopic techniques, such as time-resolved fluorescence correlation spectroscopy (FCS), could allow to follow protein changes and to analyze different aspects such as the molecular dynamics and intracellular translocation of some selected transcription factors tightly bound to the activation of the ESCs differentiation processes into neural cells. Our Results show that the pluripotency circuit is known to act as a unit that strongly represses lineage specific gene expression in ESCs. However, rather than being a monolithic entity, the pluripotency circuit components have lineage specific roles, so that the same proteins can also be used for lineage selection.