Unravelling the role of gene therapy on spinal disc mediated low back pain - PROJECT SUMMARY: Intervertebral disc degeneration (IVDD), the cause of low back pain (LBP), has been connected to a substantial socioeconomic burden, pain, and loss of function for most adults (84% globally). The significant prevalence of LBP in Nigeria and sub-Saharan Africa has compelled this study. The dysregulation of nucleus pulposus cells, the backbone's shock absorbers, and the extracellular matrix (ECM), its supporting net- work, dictates the disease progression. A type of planned cell death known as pyroptosis causes the release of specific proteins that harm the cell and its structures. In IVDD, pyroptosis accelerates cellular support system deterioration, which induces the illness. Pyroptosis may alter IVDD progression, but its exact mechanism is un- clear. MicroRNAs (miRNAs) molecules assist the cells in controlling the kinds and amounts of proteins produced. miRNAs are essential for cell formation, function, and pyroptosis regulation. The IVDD rat model is appropriate for studying the cause and treatment intervention in LBP since rats have a comparable structure and content to human discs. Fine needle piercing is preferred for developing IVDD models due to its simplicity, convenience, and high repeatability. Our team has examined miRNA’s safety and effectiveness in various aging and develop- mental conditions. However, in rat models, the impact of miR27a-3p on pyroptosis-mediated routes on spine disc cells and how it works are unknown. The rationale and central hypothesis for this project is that gene therapy (miR27a-3p) mitigates the impacts of pyroptosis proteins on ECM composition, disc cell viability, and functions in the annular puncture rat model of IVDD. In Aim 1, the research team will utilize the annular puncture rat model to investigate if IVDD activates pyroptosis proteins, influencing disc structure, function, and cellular integrity at 4, 8, and 12 weeks. Aim 2 will explore if inhibiting caspase-1 preserves ECM components, disc cell viability, and functions in the annular punctured rat model of IVDD at 4, 8, and 12 weeks. In Aim 3, the team will scrutinize if AAV6.miR27a-3p restores cell viability and functionality via CASP1/NLRP3 pathways in an annular puncture model of IVDD in rats at 4, 8, and 12 weeks. With the guidance of an exceptional mentorship team, the K43 candidate, Dr. Ogunlade, will acquire knowledge and training in 1) advanced biomedical courses, 2) research methodology and design, 3) data man- agement and analysis, and 4) manuscript writing and grantsmanship. These training goals will facilitate Dr. Ogunlade’s long-term goal to become an independent biomedical scientist focused on translational research using gene therapy intervention for LBP, build a collaborative network across different disciplines, and ultimately assemble his own research team/group. This K43 proposal will provide preliminary data and the expertise needed to design a more extensive interventional study relating to low back pain caused by IVDD rampaging among Nigerians and, by extension, to the African continent using gene therapy intervention. 1
