Effect of the modulation of the engrailed-1 in induced pluripotent stem cells on osteoblast differentiation and bone formation in defects created in mouse calvaria

• Grant number: 23/02072-0
• Support Opportunities: Regular Research Grants
• Start date: September 01, 2023
• End date: August 31, 2025
• Field of knowledge: Health Sciences - Dentistry - Oral and Maxillofacial Surgery
• Principal Investigator: Márcio Mateus Beloti
• Grantee: Márcio Mateus Beloti
• Host Institution: Faculdade de Odontologia de Ribeirão Preto (FORP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated researchers: Adalberto Luiz Rosa ; Emanuela Prado Ferraz ; Gary S Stein

Abstract

Cell therapy using stem cells derived from several tissues has been the focus of scientific investigations as a promising alternative for treating bone defects. Embryonic stem cells exhibit high plasticity and self-renewal potential, increasing their capacity of differentiating into cells of all tissues, excepting the embryonic ones. To minimize immunological aspects, and ethical and religious issues related to the use of embryonic cells, induced pluripotent stem cells (iPSs), generated by reprogramming somatic cells, have therapeutic potential to repair different tissues, including bone. 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 modulates the craniofacial development, which is underexplored in bone tissue, especially in bone repair. Thus, we established the following hypothesis: EN1 participates in the osteoblast differentiation of iPSs and in 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 iPSs, 2) evaluate the effect of En1 overexpression and silencing mediated by CRISPR/Cas9 on the osteoblast differentiation of iPSs 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 osteogenesis. (AU)