La-İlişkili Protein 7'nin Kondrogenez, Osteogenez ve Nörogenez Süreçlerindeki Fonksiyonunun Belirlenmesi

Abstract

The aim of this thesis is to elucidate the role of the LARP7 gene, in the processes of osteogenesis, chondrogenesis, and neurogenesis. Time-dependent expression of LARP7 was examined in three different differentiation models. The LARP7 expression was suppressed using RNA interference, and global gene expression profiles and alternative splicing events were investigated in this model through RNA sequencing. In the initial phase of the study, the expression levels of LARP7, 7SK RNA, and other La-related proteins were analysed. LARP7 expression was found to increase during the mid-phase of osteogenesis, suggesting a role in matrix maturation. siRNA-mediated suppression of LARP7 resulted in decreased calcium deposition and delayed osteogenesis, emphasizing its importance in bone formation. During chondrogenesis, LARP7 expression remained constant, indicating no significant role in this process. No delays or regressions were observed in chondrogenesis following LARP7 suppression. These findings suggest that skeletal anomalies observed in Alazami syndrome are primarily due to LARP7 deficiency in osteogenesis. Focused experiments on the mid-phase of osteogenesis revealed reductions in transcripts coding for extracellular matrix proteins upon LARP7 suppression. This highlights the role of LARP7 in maintaining tissue architecture and osteogenesis. The study also investigated LARP7's role in neurogenesis. Transcriptome analysis revealed changes in genes related to ribosomes and mitochondria, suggesting LARP7's role in neuronal cell metabolism and protein synthesis. Alternative splicing events were affected by LARP7 suppression, especially exon skipping, which is linked to neurodevelopmental disorders. The findings open new avenues to an understanding of congenital abnormalities in Alazami syndrome and to exploring therapeutic strategies targeting LARP7.