Study of PI3K/AKT signaling pathways in acute myeloid leukemia using selective ion monitoring (SRM).

Abstract

Acute myeloid leukemia (AML) is a group of diseases with unfavorable prognosis. During leukemogenesis several changes occur in genome, transcriptome, proteome and interactome which allow the development of leukemic cell. The leukemic cell results from deregulation in different signaling pathways, leading cells to increase proliferation, impaired cell differentiation and higher resistance to apoptosis. Among several of these altered signaling pathways, the focus of the current proposal will be the mechanisms related to the PI3K/AKT pathway, which will be studied as a model for early progression, increased proliferation and apoptosis evasion in leukemic cells. Changes described in PI3K/AKT signaling pathway may lead to a constitutive activation of downstream targets, such as AKT kinase. Our research will be based on: the role of the transmembrane adaptor protein LAT2 and the induction of apoptosis or the use of inhibitors, which alter the AKT signaling pathway in hematopoietic cells. To evaluate the regulation of this pathway, we will use targeted proteomics, and develop a panel of proteotypic peptides corresponding to tryptic fragments of relevant proteins related to PI3K/AKT pathway. This panel will be used for large scale monitoring of different samples (treated or not with kinase inhibitors and / or antineoplastic drugs, or even for specific gene knockout) using liquid chromatography coupled to mass spectrometry, referred to as SRM or MRM (selected reaction monitoring, or multiple). With this approach, we intend to obtain important functional, temporal and quantitative information about proteins in PI3K/AKT pathway using AML lineages. The identification of novel protein targets and deeper understanding of cellular signaling pathways are fundamental to the development of new drugs.