Effect of Notch1 signaling in hiPSC-derived cardiomyocytes differentiation

• Grant number: 21/14876-1
• Support Opportunities: Scholarships in Brazil - Master
• Start date: May 01, 2022
• End date: August 31, 2023
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: José Eduardo Krieger
• Grantee: Nikolas Dresch Ferreira
• Host Institution: Instituto do Coração Professor Euryclides de Jesus Zerbini (INCOR). Hospital das Clínicas da Faculdade de Medicina da USP (HCFMUSP). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

The are great prospects for cell therapy in regenerative medicine with the advancement of pluripotent stem cell studies. But, for future applications, there are important questions that need to be addressed. Observed post-transplant arrhythmia in the heart is one of the main concerns related to transplantation of cardiomyocytes derived from induced pluripotent stem cell (iPSC-CM). This outcome has been associated with the electric heterogeneity of the cell populations obtained. In this regard, many studies seek the mechanisms that different types of adult cardiac cells are formed, that include working and conduction cardiomyocytes. Recently, our group showed that inhibiting Notch1 signaling prevents the specialization of fast-conducting fibers in response to fibroblast-secreted stimuli. Hyperactivation of this pathway has also been linked to increased expression of conduction markers and a change in the electrical profile of cardiac cells. In this context, the aim of this project is to investigate whether inhibition of Notch1 activation during the hiPSCs-derived cardiomyocytes differentiation induces the specialization of working cells and whether this effect is abolished after the inhibition is removed. For this purpose, hiPSCs will be submitted to the cardiomyocyte differentiation protocol and will be treated for 3 or 10 days with DAPT (a Notch1 signaling inhibitor). After the protocol, electrical and contractile function, gene expression and phenotypic characterization of all cells will be evaluated. It is worth mentioning that the opportunity to work with hiPSC differentiation in working cells is unprecedented and will be crucial for the validation of the phenotype found by our laboratory, cooperating with the development of cell therapies. In addition, it will help us to better understand the main mechanisms of the involvement of the conduction system in some heart diseases, such as arrhythmias. It should also be noted that hiPSC colonies and the differentiation protocols are already established in our laboratory, and the hiPSC-CMs have already been characterized. (AU)