Speaker
Description
Human somatic neural stem cells (NSCs) are emerging as promising advanced therapy medicinal products for treating neurodegenerative and demyelinating disorders. However, challenges remain, including the large-scale production of donor cells under GMP conditions and the necessity of immunosuppressive regimens in allogeneic transplant settings. Human induced pluripotent stem cell-derived neural stem/progenitor cells (hiPSC-NSCs) offer a potential alternative but comprehensive studies investigating their identity and safety are still limited. Significantly, the hostile environment of demyelinating conditions, such as Multiple Sclerosis, could negatively impact transplanted NSC, highlighting the need for innovative approaches to enhance their therapeutic potential.
We performed genome-wide omics analyses, including RNA sequencing (RNA-seq), chromatin immunoprecipitation sequencing (ChIP-seq), and single-cell RNA sequencing (scRNA-seq), to compare hiPSC-NSCs with hiPSCs, human fetal NSCs (hfNSCs), and glioblastoma stem cells (GSCs) to examine their transcriptional and epigenetic signatures and assess their NSC identity and safety profile. Our analysis revealed that hiPSC-NSCs acquired a transcriptomic profile that represents an intermediate middle-radial glia (MRG) and late-radial glia (LRG) state, resembling primitive ventral RG (vRG) isolated from the human fetal brain. Notably, hiPSC-NSCs share key transcriptional and epigenetic characteristics with hfNSCs while significantly diverging from GSCs. Long-term transplantation in immunodeficient mice showed robust and stable engraftment of hiPSC-NSCs, with predominant differentiation into glial cells and no tumor formation. We are investigating whether hiPSC-NSCs engineered through CRISPR activation (CRISPRa) to overexpress therapeutic genes can promote oligodendrocyte maturation, immunomodulation, and neuroprotection. This approach aims to counteract the hostile environment of Multiple Sclerosis and address the remyelination failure.
These findings offer valuable transcriptional and epigenetic reference datasets that aid in defining the maturation stage of NSCs derived from different hiPSC sources. Additionally, the results demonstrate the long-term safety of hiPSC-NSCs. The potential to enhance their therapeutic effects through CRISPRa further underscores their viability as an alternative to hfNSCs for clinical applications.
| Author(s) | *Marco Luciani 1, Chiara Garsia1,2, Stefano Beretta1,2, Ingrid Cifola3, Clelia Peano4,5, Ivan Merelli1, Luca Petiti3, Annarita Miccio6, Vasco Meneghini1,2, Angela Gritti1,2 |
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| Affiliation(s) | "1. San Raffaele Telethon Institute for Gene Therapy, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) San Raffaele Scientific Institute, Milan, Italy 2. Vita-Salute San Raffaele University, Milan, Italy 3. Institute for Biomedical Technologies (ITB), National Research Council (CNR), via F.lli Cervi 93, 20054 Segrate, Milan, Italy 4. Institute of Genetics and Biomedical Research, UoS of Milan, National Research Council, Rozzano, Milan, Italy 5. Human Technopole, Via Rita Levi Montalcini 1, Milan, Italy 6. IMAGINE Institute, Université de Paris, Sorbonne Paris Cité, Paris, France" |
