RRC ID 77293
著者 Pavani G, Laurent M, Fabiano A, Cantelli E, Sakkal A, Corre G, Lenting PJ, Concordet JP, Toueille M, Miccio A, Amendola M.
タイトル Ex vivo editing of human hematopoietic stem cells for erythroid expression of therapeutic proteins.
ジャーナル Nat Commun
Abstract Targeted genome editing has a great therapeutic potential to treat disorders that require protein replacement therapy. To develop a platform independent of specific patient mutations, therapeutic transgenes can be inserted in a safe and highly transcribed locus to maximize protein expression. Here, we describe an ex vivo editing approach to achieve efficient gene targeting in human hematopoietic stem/progenitor cells (HSPCs) and robust expression of clinically relevant proteins by the erythroid lineage. Using CRISPR-Cas9, we integrate different transgenes under the transcriptional control of the endogenous α-globin promoter, recapitulating its high and erythroid-specific expression. Erythroblasts derived from targeted HSPCs secrete different therapeutic proteins, which retain enzymatic activity and cross-correct patients' cells. Moreover, modified HSPCs maintain long-term repopulation and multilineage differentiation potential in transplanted mice. Overall, we establish a safe and versatile CRISPR-Cas9-based HSPC platform for different therapeutic applications, including hemophilia and inherited metabolic disorders.
巻・号 11(1)
ページ 3778
公開日 2020-7-29
DOI 10.1038/s41467-020-17552-3
PII 10.1038/s41467-020-17552-3
PMID 32728076
PMC PMC7391635
MeSH Animals CRISPR-Cas Systems / genetics Cell Engineering / methods* Cell Line Female Gene Editing* Gene Expression Regulation Hematopoietic Stem Cell Transplantation / methods* Hematopoietic Stem Cells / metabolism* Hemophilia A / therapy Humans Metabolic Diseases / therapy Mice Promoter Regions, Genetic / genetics Transplantation, Autologous / methods Transplantation, Heterologous alpha-Globins / genetics alpha-Globins / metabolism
IF 12.121
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リソース情報
ヒト・動物細胞 HUDEP-2(RCB4557)