Immunocompetent 3D human CNS Models: a novel approach for preclinical development of gene therapies

Abstract

"Recombinant adeno-associated virus (rAAV) gene therapies hold significant promise for treating central nervous system (CNS) disorders. However, their effectiveness in clinical settings is often limited by the immune responses mounted by the host. Astrocytes and microglia, the resident immune cells of the CNS, play a crucial role in initiating innate immune responses, which can lead to neuroinflammation. However, a comprehensive understanding of the molecular mechanisms driving and sustaining glial cell activation remains unclear due to the lack of human models illustrating the neuro-immune axis. To accelerate the clinical use of rAAVs, thorough preclinical evaluation of their efficacy and safety in human models is essential. The in vitro replication of CNS phenotypic and functional characteristics requires advanced culture techniques to closely emulate the structural and molecular complexity of in vivo systems. Models derived from the differentiation of human induced pluripotent stem cells (hiPSCs) in a three-dimensional (3D) environment hold potential as complementary tools for animal-based preclinical research. This thesis aims to create innovative human 3D CNS models that effectively replicate the key features of neuroinflammation by integrating hiPSC technology with stirred-tank bioreactor systems and utilizing a broad array of characterization methods. The models developed for advancing preclinical research of ATMPs, specifically viral-based gene therapies, demonstrated significant potential.(...)"