Micro Ribonucleic Acid−29a (miR−29a) Antagonist Normalizes Bone Metabolism in Osteogenesis Imperfecta (OI) Mice Model

Abstract

Osteogenesis imperfecta (OI) is not curative nowadays. This study tried to unriddle the therapeutic potential of micro ribonucleic acid-29a (miR-29a) antagonist in treating OI in a mouse animal model (B6C3Fe a/a-Col1a2oim/J). We showed that the expression levels of miR-29a were higher in bone tissues obtained from the OI mice than from wild-type mice demonstrated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and in situ hybridization assay. We established lentivirus-shuttled vector expressing miR-29a antisense oligonucleotide (miR-29a-AS) and miR-29a precursors (pre-miR-29a), showing that the inferior bony architecture in micro-computed tomography and pertinent morphometric parameters could be rescued by miR-29a-AS and deteriorated by pre-miR-29a. The decreased proliferating cell nuclear antigen (PCNA), increased Dickkopf-1 (DKK1), and decreased β-catenin expression in OI mice could be accentuated by pre-miR-29a and normalized by miR-29a-AS. The decreased osteogenesis and increased osteoclastogenesis in OI mice could also be accentuated by pre-miR-29a and normalized by miR-29a-AS. miR-29a-AS did not seem to possess severe hepatic or renal toxicities.

Dickkopf-1 (DKK1) blockade mitigates osteogenesis imperfecta (OI) related bone disease

“…Notably, serum samples from patients with elevated DKK1 levels hindered osteoblast differentiation and the expression of OPG in vitro (Brunetti et al 2016 ). In our prior investigation into the therapeutic potential of micro ribonucleic acid-29a (miR-29a) antagonists for treating OI in a murine animal model, we demonstrated the ability of pre-miR-29a to augment the local expression of DKK1 in OI-afflicted mice, which was subsequently restored to normal levels by anti-sense miR-29a (Ko et al 2023a ). These observations align intriguingly with the current study’s exploration into the therapeutic ramifications of DKK1-AS in OI mice.…”

“…The assessment of gene transcription levels was conducted employing the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay following our previous works (Ko et al 2022a , b , 2023a , b ). Tissues were pulverized under liquid nitrogen in the absence of RNase, and total RNA was extracted using the RiboPure RNA purification kit (Thermo Fisher Scientific, Waltham, MA, USA).…”

“…Calcium mass deposits were stained black by the kit, and the area of von Kossa-stained mineralized matrices (mm 2 /field) in each × 125 magnification field was measured using light microscopy and image analysis software (Carl Zeiss). Six fields were randomly selected from each well, with a total of six wells for each treatment, to quantify the von Kossa-stained area (Ko et al 2023a ).…”

“…Osteogenesis imperfecta (OI) is a rare hereditary bone disorder characterized by severe osteoporosis, deformity, fragility, and recurrent fractures (Ko et al 2023a ; Syu et al 2022 ; Faienza et al 2023 ; Tyurin et al 2022 ). Mutations in the genes encoding type I collagen, abnormalities in collagen folding, or post-transcriptional modifications that disrupt the ultrastructure of collagen in the skeletal microenvironment are implicated in the pathogenesis of OI (Barnes et al 2010 , 2006 ).…”

Changes in Serum Bone Metabolism Markers after Living Donor Liver Transplantation (LDLT) and Their Association with Fracture Occurrences

“…No apparent intra-group differences in RANKL (Figure 13) and OPG/RANKL ratio (Figure 14) were noted (p = 0.744 and p = 0.491, respectively). Dickkopf-related protein 1 (DKK1) is a strong skeletal-deleterious factor [17]. DKK1 is a secreted protein that functions as an inhibitor of the canonical Wnt signaling pathway.…”