Effect of Engrailed-1 protein modulation in mesenchymal stem cells on osteoblastic differentiation and bone formation in defects created in mouse calvaria

• Grant number: 23/00538-2
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: September 01, 2023
• End date: August 31, 2027
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Márcio Mateus Beloti
• Grantee: Robson Diego Calixto
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 17/12622-7 - Cell therapy: potential of mesenchymal stem cells, VEGF-A and BMP-9 to regenerate bone tissue, AP.TEM
• Associated scholarship(s): 25/01045-5 - Effect of the interaction between Engrailed-1 and the TGFB/SMAD3 circuit in osteoblastic differentiation of mesenchymal stem cells, BE.EP.DR

Abstract

Cell therapy using mesenchymal stem cells derived from several tissues has been the focus of scientific investigations as a promising alternative for treating bone defects. MSCs derived from adipose tissue (MSCs-AT), when aspirated and isolated, present themselves as a safe source of cells for regenerative medicine and cell therapy, since through their plasticity and angiogenic role, they can modulate the immune and anti-inflammatory response and contribute to the regeneration of bone tissue. The process of bone formation is regulated by a plethora of molecules and signalling pathways, including the protein called engrailed-1 (EN1), a transcription factor that modulate the craniofacial development, which is underexplored in bone tissue, especially in bone repair. Thus, we stablished the following hypothesis: EN1 participates of the osteoblast differentiation of iMSCs-AT and of the bone repair induced by these cells. To test this hypothesis, the aims of this study are to: 1) characterize the En1 during the osteoblast differentiation of iMSCs-AT, 2) evaluate the effect of En1 overexpression and silencing mediated by CRISPR/Cas9 on the osteoblast differentiation of iMSCs-AT and 3) evaluate the effect of of En1 overexpression and silencing mediated by CRISPR/Cas9 on the bone repair of mice calvarial defects. The results of this project will be scientifically and therapeutically relevant as EN1 can be a target protein for the development of new approaches to favour bone regeneration, by regulating events related to the osteogenesis. (AU)