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Nanoengineered 3D culture substrate enables superior persistence and polyclonal engraftment of genetically engineered hematopoietic stem cells

Title: Nanoengineered 3D culture substrate enables superior persistence and polyclonal engraftment of genetically engineered hematopoietic stem cells
Authors: Midena, Federico; Alessandrini, Laura; Conci, Claudio; Barcella, Matteo; Gazzo, Francesco; Jacchetti, Emanuela; Carsana, Edoardo; Benedicenti, Fabrizio; Vacca, Roberta; della Volpe, Lucrezia; Arévalo, Sergio; Giannetti, Kety; Barozzi, Dafne; Franchino, Martina; Zonari, Erika; Ferrua, Francesca; Farina, Giacomo; Brombin, Chiara; Cugnata, Federica; Fiumara, Martina; Tavella, Teresa; Cherubin, Leonardo; Fraschetta, Federico; Cerullo, Giulio; Osellame, Roberto; Radrizzani, Marina; Ferrari, Samuele; Merelli, Ivan; Gentner, Bernhard; Scielzo, Cristina; Brendolan, Andrea; Naldini, Luigi; Aiuti, Alessandro; Montini, Eugenio; Raimondi, Manuela T.; Di Micco, Raffaella
Contributors: Midena, Federico; Alessandrini, Laura; Conci, Claudio; Barcella, Matteo; Gazzo, Francesco; Jacchetti, Emanuela; Carsana, Edoardo; Benedicenti, Fabrizio; Vacca, Roberta; Della Volpe, Lucrezia; Arévalo, Sergio; Giannetti, Kety; Barozzi, Dafne; Franchino, Martina; Zonari, Erika; Ferrua, Francesca; Farina, Giacomo; Brombin, Chiara; Cugnata, Federica; Fiumara, Martina; Tavella, Teresa; Cherubin, Leonardo; Fraschetta, Federico; Cerullo, Giulio; Osellame, Roberto; Radrizzani, Marina; Ferrari, Samuele; Merelli, Ivan; Gentner, Bernhard; Scielzo, Cristina; Brendolan, Andrea; Naldini, Luigi; Aiuti, Alessandro; Montini, Eugenio; Raimondi, Manuela T.; Di Micco, Raffaella
Publication Year: 2026
Collection: RE.PUBLIC@POLIMI - Research Publications at Politecnico di Milano
Subject Terms: 3D culture; Wiskott-Aldrich syndrome; clonal tracking; genome editing; hematopoietic stem cells; lentiviral gene transfer; mechanobiology
Description: Ex vivo culture of hematopoietic stem and progenitor cells (HSPCs) is required for gene therapy applications but inadvertently triggers detrimental cellular responses, potentially threatening clinical success. In this study, we employ nichoids, biocompatible 3D culture substrates with cell-scale resolution, to provide HSPCs with mechanical support during ex vivo manipulation. This innovative 3D system improves HSPC multi-lineage differentiation and engraftment capacity by leveraging mechanobiological control over nuclear morphology, cytoskeleton organization, metabolism, and DNA integrity. Notably, 3D culture enables efficient genetic engineering across multiple platforms, including long-range gene editing, base- and prime-editing, and lentiviral-mediated gene addition. Moreover, this scaffold increases the clonal output and persistence of genetically engineered cells in xenotransplantation experiments, including a clinical protocol for lentiviral gene addition in Wiskott-Aldrich syndrome. Overall, we propose a transformative approach to enhance the efficacy and safety of emerging and established hematopoietic stem cell-based gene therapy applications.
Document Type: article in journal/newspaper
Language: English
Relation: info:eu-repo/semantics/altIdentifier/pmid/41512872; info:eu-repo/semantics/altIdentifier/wos/WOS:001685656000001; volume:33; issue:2; firstpage:217; lastpage:232; numberofpages:16; journal:CELL STEM CELL; https://hdl.handle.net/11311/1308797
DOI: 10.1016/j.stem.2025.12.016
Availability: https://hdl.handle.net/11311/1308797; https://doi.org/10.1016/j.stem.2025.12.016
Rights: info:eu-repo/semantics/openAccess
Accession Number: edsbas.94032C3F
Database: BASE