@misc{10481/106504,
year = {2025},
month = {4},
url = {https://hdl.handle.net/10481/106504},
abstract = {To improve ex vivo gene therapy strategies involving hematopoietic stem and progenitor cells (HSPCs), we propose a novel knock-in strategy
(named KI-Ep) aiming to achieve transgene regulation of the inserted cassette through the acquisition of naturally occurring epigenetic marks.
Based on this hypothesis, we selected CX3CR1 (a myeloid-specific gene presenting a poised histone signature on primitive HSPCs) as safe
harbor to generate KI-Ep HSPCs. We demonstrated that, unlike the expression pattern achieved with lentiviral vectors (LVs), the insertion of
a constitutive expression cassette into the intron 1 of the CX3CR1 locus (CX3CR1-I) in HSPCs resulted in very low expression levels in the
more primitive HSPCs but, crucially, strong expression in HSPC-differentiated populations (especially myeloid cells), both in vitro and in vivo.
Furthermore, we showed that the promoter of the expression cassette inserted into CX3CR1-I acquired epigenetic marks associated with
poised genes during the HSPC stage. These marks transitioned to activated histone states upon KI-Ep HSPCs differentiation. In summary, here,
we introduce the KI-Ep concept which enables the epigenetic modulation of the inserted transgene during the HSPCs stem cell stages and its
subsequent activation upon differentiation.},
organization = {Fondo Europeo de Desarrollo Regional (FEDER), NextGenerationEU, Instituto de Salud Carlos III (ISCIII): [PI18/00337, PI21/00298,  PI21/00875]},
organization = {RICORS-TerAv [RD21/ 0017/0004; RD21/0017/0027]},
organization = {FI19/00163},
organization = {FEDER/European Cohesion Fund (FSE) Andalucía [PECART-0031-2020]},
organization = {Junta de Andalucía PEER-0286-2019, EMERGIA20_00049},
organization = {Pompe Competitive Research SPARK-2022-002},
organization = {Ministerio de Ciencia e innovación (MICIN) PID2021-125077OB-C22, PLEC2021-008094},
organization = {Centro para el Desarrollo Tecnológico Industrial (CDTI), European Union-Next Generation EU [00123009/SNEO-20191072,  PMPTA22/00060]},
organization = {Spanish National Research Agency [AEI; RYC2021-031920-I, CNS2022-136033]},
organization = {Horizon Europe Framework Program X-PAND 101070950},
organization = {European Cooperation in Science and Technology (COST) [GeneHumdi-CA21113]},
organization = {Instituto de Salud Carlos III (ISCIII) [RD21/ 0017/0004]},
organization = {Universidad de Granada/CBUA},
publisher = {Oxford University Press},
title = {Donor insertion into CX3CR1 allows epigenetic modulation of a constitutive promoter on hematopoietic stem cells and its activation upon myeloid differentiation},
doi = {10.1093/nar/gkaf344},
author = {Ramos Hernández, Iris and Fuster-García, Carla and Aguilar González, Araceli and Lozano-Vinagre, María L. and Guenechea-Amurrio, Guillermo and Sánchez-Luque, Franscisco J. and Gonçalves, Manuel A.F.V. and Cathomen, Toni and Muñoz, Pilar and Molina-Estévez, Francisco Javier and Martín Molina, Francisco},
}