| RRC ID |
77293
|
| Author |
Pavani G, Laurent M, Fabiano A, Cantelli E, Sakkal A, Corre G, Lenting PJ, Concordet JP, Toueille M, Miccio A, Amendola M.
|
| Title |
Ex vivo editing of human hematopoietic stem cells for erythroid expression of therapeutic proteins.
|
| Journal |
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.
|
| Volume |
11(1)
|
| Pages |
3778
|
| Published |
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
|
| Resource |
| Human and Animal Cells |
HUDEP-2(RCB4557) |
