Maximizing Safety and Efficacy in Hematopoietic Stem Cell Gene Therapy

Hematopoietic stem cell (HSC) gene therapy has been recently successfully employed for certain rare blood disorders including hemoglobinopathies, which resulted even in marketing authorizations such as Lyfgenia™ and Casgevy™. Despite these advancements, the appearance of leukemic events in the case of Sickle Cell Disease combined with the therapy's high price render the wider access, and hence the overall applicability of this approach, doubtful. Therefore to make HSC gene therapy more widely applicable, an effort is required toward robust and less laborious manufacturing processes that ensure reproducibility and predictability to maximize the safety as well as providing the capacity to generate engineered HSCs at large scale. Automation is the key to achieve these goals and, to this end, the CliniMACS Prodigy® as an automated, closed, GMP-compliant platform for HSC transduction is an ideal candidate to streamline cell processing. To this end, the scope of this project is a) to provide alternative methodology for genetic engineering of CD34⁺ cells by testing several transduction enhancers to achieve a clinically translatable transduction efficiency both in the manual process (open steps) and in large scale on the CliniMACS Prodigy and b) to assess the safety of both approaches. In our first line of experiments, comparable cell recovery and viability between the small and large scale conditions were observed, however superior performance of the automated platform was repeatedly shown in terms of transduction efficiency compared to open manual steps. Additionally, significantly higher transduction efficiency was achieved in the presence of transduction enhancers, preserving cell recovery and viability, without altering the stem cell clonogenic capacity. Based on these results, we propose a combined strategy of transduction enhancers on the automated platform to achieve significantly higher transduction efficiency. Ongoing experiments investigate the mechanisms involved in better performance of lentiviral transduction on the automated platform with molecular analysis via Next Generation Sequencing (NGS) and respective bioinformatics analysis.

Giommetti:Miltenyi Biotec B.V. & Co. KG: Current Employment. Barra:Miltenyi Biotec B.V. & Co. KG: Current Employment. Bosbach:Miltenyi Biotec B.V. & Co. KG: Current Employment. Olevska:Miltenyi Biotec B.V. & Co. KG: Current Employment. Dzionek:Miltenyi Biotec B.V. & Co. KG: Current Employment. Soltenborn:Miltenyi Biotec B.V. & Co. KG: Current Employment. Hebbeker:Miltenyi Biotec B.V. & Co. KG: Current Employment. Oberbörsch:Miltenyi Biotec B.V. & Co. KG: Current Employment. Martinez Carrera:Miltenyi Biotec B.V. & Co. KG: Current Employment. Jelveh:Miltenyi Biotec B.V. & Co. KG: Current Employment. Bisdorf:Miltenyi Biotec B.V. & Co. KG: Current Employment. Matzke:Miltenyi Biotec B.V. & Co. KG: Current Employment. Khorkova:Miltenyi Biotec B.V. & Co. KG: Current Employment. Wöhle:Miltenyi Biotec B.V. & Co. KG: Current Employment. Tomiuk:Miltenyi Biotec B.V. & Co. KG: Current Employment. Bosio:Miltenyi Biotec B.V. & Co. KG: Current Employment. Papanikolaou:Miltenyi Biotec B.V. & Co. KG: Current Employment.