Author(s): Brooklyn Hopkins
Mentor(s): Christina Jacobsen
Institution BYU
Osteogenesis Imperfecta (OI) is a genetically heterogeneous disorder primarily caused by mutations in the COL1A1 or COL1A2 genes. This leads to an impaired production of osteoblasts, resulting in fragile, brittle bones or improper bone formation in children. The purpose of this study is to more fully understand the mechanisms behind the impairment of growth during transformation of chondrocytes to osteoblasts in patients with OI. This study investigates the mechanisms underlying growth impairment during the transformation of chondrocytes into osteoblasts. Growth plate-like chondrocytes were first isolated, and the COL1A1 gene was subsequently knocked out to create the knockout (KO) cell line. Both the KO and a non-targeting (NT) control cell line were plated in micromass at a density of 20 million cells per mL, 20 uL per micromass, and cultured in chondrocyte media for 14 days. Three micromasses were harvested per time point. KO vs Control micromasses were then profiled by qPCR for expression of genes typically associated with different states of chondrocyte maturation as well as osteoblast markers. The results demonstrate that cells lacking Col1A1 expression exhibited increased gene expression for both chondrocyte (Col10) and osteoblast (BGLAP) markers, suggesting enhanced maturation of osteoblasts in the absence of type 1 collagen. These results suggest that COL1A1 is important in keeping a balance between bone and cartilage development. However, as this experiment is currently being replicated to verify the consistency and reproducibility of these outcomes, further studies will be necessary to better understand the mechanisms underlying this relationship.