First primary adult sheep neural screen identifies therapeutic candidates for spinal cord injury

ABSTRACT: High content screening efforts for age-associated neurological disorders previously relied on embryonic, early neonatal, or induced pluripotent stem cell-derived neural cells. This dichotomy in age between cells in drug screens and the target adult population in the clinic likely impedes translational successes. Aging reduces regenerative potential, impeding glial process extension and neuronal neurite regeneration. Pharmacologically enhancing the extension of processes in adult glia and neurite outgrowth in adult neurons holds therapeutic value. However, no previous assay allowed for high content screening in primary adult neural cells. We previously developed a screening platform that uses primary adult neurons from rodents. Though, the limited yield in neural cells prevented true high content screening. We recently adapted this brain tissue processing method to larger mammals. It led to the development of a high content screen using primary adult sheep neural cells and the first recorded case of primary adult sheep neurons being cultured, as demonstrated by immunocytochemistry, western blotting, and single-nucleus RNA sequencing. We conducted a pilot screen of 2281 unique compounds from preselected libraries, identifying numerous compounds that have shown efficacy in the clinic for various neurological indications, validating our protocol. We identified for the first time several new molecules promoting processes/neurite growth in adult cells, including GSK180736A, PHA665752, and StemRegenin1. All three compounds were subsequently tested in a preclinical spinal cord injury model to identify neurologically beneficial hits. Altogether, we provide here a novel drug screening tool that may help identify new leads for age-associated neurological disorders.

ORGANISM(S): Ovis aries

PROVIDER: GSE333863 | GEO | 2026/06/15

REPOSITORIES: GEO