Modeling of monogenic diseases for physiopathological studies and pharmacological tests using specialized cells derived from iPSCs

Abstract

Currently, one of the great challenges in medical research is to model cells in vitro capable of recapitulating physiological processes in the most similar way possible to complex organisms. The difficulty of obtaining specialized cells for studies in laboratories such as cardiomyocytes, endothelial cells and neurons has always been a challenge for the development of more accurate studies, which has been overcome in the last decade with the use of Induced Pluripotent Stem Cells (iPSCs). Since its development, iPSCs have shown their potential to differentiate into specialized cells. In addition, these cells behave like embryonic stem cells, bypassing ethical controversies, with no sample limit and with the ability to differentiate into almost all cell types. In this project we aim to use these cells in order to create an in-vitro platform for pathophysiological studies and drug analysis in 3 monogenic diseases. For this purpose, the development of cardiomyocytes and neurons derived from iPSCs with changes in genes responsible for the development of Fabry's Disease (DF) and Long QT Syndrome Type 3 (LQT3) was proposed. In addition, iPSC-derived endothelial cells carrying mutations responsible for the development of Hereditary Angioedema (HA) will also be developed. All specialized cells will be modeled from adult peripheral blood cells of patients with monogenic diseases due to or mutations induced by the CRISPr-Cas9 system. For this, iPSCs will be generated through reprogramming with episomal vectors and differentiated with specific factors. The differentiated cells will be characterized, and later analyzed in their pathophysiological processes, through molecular, fluorescent markers and electrophysiological analyzes. The use of these cells for the development of organoids may also better elucidate the biological processes involved in the accumulation of GB3 in DF, and their relationship with neural and cardiac involvement, in addition to the electrophysiological processes in LQT3 and vascular permeability in HA. The platform can also open perspectives for drug tests and bring important pharmacogenomic information capable of more assertively guiding drug prescriptions according to the type of mutation presented by the patient. (AU)