Analysis of mitocan alpha-tocoferol mitochondrial action: cellular model for acute myeloid leukemia carrying the FLT3-ITD mutation

• Grant number: 17/02324-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2017
• End date: May 31, 2018
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Eduardo Magalhães Rego
• Grantee: Adriane Souza Lima
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Acute myeloid leukemia (AML) can be defined, in most cases, as a cluster of diseases of a sporadic nature and represents failure in the maturation of myeloid cells, with the expansion of the immature lineage in blood, marrow and other tissues. Of the many genetic changes that can result in the disease, the mutation in the FLT3 gene in the form of internal tandem duplications (ITD) appears in about 30% of the cases and is linked to a poor prognosis, with lower chances of remission and higher rates of relapse. Among the new drugs investigated for the treatment of AML with FLT3-ITD, the tyrosine kinase inhibitors stand out. Quisartinib and midostaurin, for example, showed substantial antitumor action with significant clinical results. However, they also demonstrated that when used as monotherapy, these drugs are unable to maintain or even achieve complete remission. Thus, investigations of new classes of drugs, possibly used in combined therapy with classic chemotherapeutic agents or with tyrosine kinase inhibitors are highly relevant. One of these new classes are the mitocans, a group of drugs capable of acting in the mitochondria, interrupting the production of energy by the cell, increasing the production of reactive oxygen species and activating the intrinsic chain of apoptosis. In this study, mitocan alpha-tocopherol is used in the treatment of MV4,11 cells, a model line for AML with FLT3-ITD, in order to demonstrate its mitochondrial action in leukemic cells, potentializing its chance of future use in the treatment of the disease in question. Thus, the main objectives of the project are: 1) To find the effective dose (ED 50) of mitocan alpha-tocopherol resulting in apoptosis of MV cells 4,11. 2) To analyze the mitochondrial membrane potential of MV4,11 cells treated with mitocan alpha-tocopherol to evaluate the hypothesis that the pro-apoptotic effect of alpha-tocopherol in MV4,11 tumor cells is due to mitochondrial membrane depolarization. 3) To analyze activation of caspases 3 and 9 by the Western Blot method in MV4,11 cells treated with alpha-tocopherol. 4) To analyze the phosphorylation of the FLT3 tyrosine kinase receptor in alpha-tocopherol treated MV4,11 cells. (AU)