Drug toxicity detection by fluorescent reporters in cardiohepatic co-culture system

• Grant number: 23/18178-2
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: May 01, 2024
• End date: February 29, 2028
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Mayana Zatz
• Grantee: Sabrina Kaori Kadowaki Komatsu
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Developing new drugs is slow and expensive, especially because the assessment of cardiac and hepatic toxicity in preclinical trials frequently leads to compound rejection. Toxicity assessment primarily requires animal models, which are physiologically distinct from humans. To enhance the accuracy of drug screening, cardiomyocytes and hepatocytes derived from human induced pluripotent stem cells (hiPSCs) represent an alternative to traditional animal models. However, the lack of crosstalk between cell types in isolated cell models is imprecise for toxicity assessment of secondary metabolites. Microfluidic platforms allow the co-culture of hepatic and cardiac cells and reproduce the vascular system through directional media flow in a high-throughput scale, compatible with the pharmaceutical industry standards. To detect toxicity, generating gene edited cells with conditional fluorescent markers provides a precise and automated alternative to traditional colorimetric and luminescent assays. Therefore, the aim of this project is to test a microfluidic device containing genetically edited hiPSC-derived hepatocytes and cardiomyocytes to report drug-induced apoptosis through fluorescence. We will generate transgenic hiPSCs through insertion of caspase 3 cleavage-dependent FRET protein reporters, induce primary and secondary toxicity using commercial drugs, and assess apoptosis through immunofluorescence and ELISA. (AU)