ALSTaR Cells - Novel Stem-cell-based Therapy for Amyotrophic Lateral Sclerosis: A Research Protocol

Abstract

Introduction: Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disease that results in the loss of motor neurons leading to limb paralysis and eventual death. Stem cell transplantation can be used to replenish the atrophied motor neurons and slow the progression of the disease, while the use of biomaterials, genetic engineering, and nanoparticles can reduce the hostility of the microenvironment. Thus, we propose a novel combinatorial therapeutic approach termed Amyotrophic Lateral Sclerosis Therapeutic and Regenerative (ALSTaR) cell treatment. Methods: This unified strategy will employ miRNA-124 and chitosan polyplex biomaterial to subsequently enhance engraftment of neural progenitor cells (NPCs), which will be engineered to overexpress an autophagy-regulatory gene, TFEB, and secrete an autophagy-inducing drug, trehalose. We will bioengineer an organoid model of the ventral column of the spinal cord which will be used for extensive in vitro characterization of ALSTaR cells through single-slice electrophysiology and immunocytochemistry, followed by in vivo work. Using the SOD1G93A mutant mouse model, weekly behavioural assessments of motor recovery will be conducted until the humane endpoint; thereafter extensive immunohistochemical and protein quantification analyses will be performed to determine treatment efficacy. Results: ALSTaR cell treatment should result in motor functional recovery and lifespan elongation through several cellular and molecular mechanisms, including decreased neuroinflammation, upregulation of autophagy, degradation of protein aggregates, enhancement of transplanted cell integration, and regeneration of axons. Discussion: In vitro characterization of ALSTaR cells in bioengineered spinal cord organoids will reveal stable electrophysiological recordings of motor neurons, higher levels of neuronal differentiation markers, and lower levels of inflammation markers in the ALSTaR group compared to the control group or the groups treated with just the biomaterial or just the NPCs, suggesting neuronal recovery. Behavioural assessments in the ALS mouse model will reveal increased motor coordination, neuromuscular strength, and motor activity in the ALSTaR group compared to other groups, suggesting motor functional recovery. In vivo characteriz ation of ALSTaR cells in the ALS mouse model will suggest enhanced stem cell integration and recovery of cellular and molecular processes. Conclusion: With currently no effective treatment for ALS, this novel combinatorial treatment strategy could improve the health-related quality of life of patients suffering from this debilitating disease.