PMID- 36206730
OWN - NLM
STAT- MEDLINE
DCOM- 20221011
LR  - 20221118
IS  - 1875-9777 (Electronic)
IS  - 1934-5909 (Print)
IS  - 1875-9777 (Linking)
VI  - 29
IP  - 10
DP  - 2022 Oct 6
TI  - Choice of template delivery mitigates the genotoxic risk and adverse impact of 
      editing in human hematopoietic stem cells.
PG  - 1428-1444.e9
LID - S1934-5909(22)00378-2 [pii]
LID - 10.1016/j.stem.2022.09.001 [doi]
AB  - Long-range gene editing by homology-directed repair (HDR) in hematopoietic 
      stem/progenitor cells (HSPCs) often relies on viral transduction with recombinant 
      adeno-associated viral vector (AAV) for template delivery. Here, we uncover 
      unexpected load and prolonged persistence of AAV genomes and their fragments, 
      which trigger sustained p53-mediated DNA damage response (DDR) upon recruiting 
      the MRE11-RAD50-NBS1 (MRN) complex on the AAV inverted terminal repeats (ITRs). 
      Accrual of viral DNA in cell-cycle-arrested HSPCs led to its frequent 
      integration, predominantly in the form of transcriptionally competent ITRs, at 
      nuclease on- and off-target sites. Optimized delivery of integrase-defective 
      lentiviral vector (IDLV) induced lower DNA load and less persistent DDR, 
      improving clonogenic capacity and editing efficiency in long-term repopulating 
      HSPCs. Because insertions of viral DNA fragments are less frequent with IDLV, its 
      choice for template delivery mitigates the adverse impact and genotoxic burden of 
      HDR editing and should facilitate its clinical translation in HSPC gene therapy.
CI  - Copyright (c) 2022 The Author(s). Published by Elsevier Inc. All rights reserved.
FAU - Ferrari, Samuele
AU  - Ferrari S
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy; Vita-Salute San Raffaele University, Milan 20132, 
      Italy.
FAU - Jacob, Aurelien
AU  - Jacob A
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Cesana, Daniela
AU  - Cesana D
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Laugel, Marianne
AU  - Laugel M
AD  - INSERM UMR 1089, University of Nantes, CHU of Nantes, Nantes 44200, France.
FAU - Beretta, Stefano
AU  - Beretta S
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Varesi, Angelica
AU  - Varesi A
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Unali, Giulia
AU  - Unali G
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Conti, Anastasia
AU  - Conti A
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Canarutto, Daniele
AU  - Canarutto D
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy; Vita-Salute San Raffaele University, Milan 20132, 
      Italy; Pediatric Immunohematology Unit and BMT Program, IRCCS San Raffaele 
      Scientific Institute, Milan 20132, Italy.
FAU - Albano, Luisa
AU  - Albano L
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Calabria, Andrea
AU  - Calabria A
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Vavassori, Valentina
AU  - Vavassori V
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Cipriani, Carlo
AU  - Cipriani C
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Castiello, Maria Carmina
AU  - Castiello MC
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy; Institute for Genetic and Biomedical Research (UOS 
      Milan Unit), National Research Council, Milan 20132, Italy.
FAU - Esposito, Simona
AU  - Esposito S
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Brombin, Chiara
AU  - Brombin C
AD  - University Center for Statistics in the Biomedical Sciences, Vita-Salute San 
      Raffaele University, Milan 20132, Italy.
FAU - Cugnata, Federica
AU  - Cugnata F
AD  - University Center for Statistics in the Biomedical Sciences, Vita-Salute San 
      Raffaele University, Milan 20132, Italy.
FAU - Adjali, Oumeya
AU  - Adjali O
AD  - INSERM UMR 1089, University of Nantes, CHU of Nantes, Nantes 44200, France.
FAU - Ayuso, Eduard
AU  - Ayuso E
AD  - INSERM UMR 1089, University of Nantes, CHU of Nantes, Nantes 44200, France.
FAU - Merelli, Ivan
AU  - Merelli I
AD  - Institute for Biomedical Technologies, National Research Council, Segrate 20090, 
      Italy.
FAU - Villa, Anna
AU  - Villa A
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy; Institute for Genetic and Biomedical Research (UOS 
      Milan Unit), National Research Council, Milan 20132, Italy.
FAU - Di Micco, Raffaella
AU  - Di Micco R
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Kajaste-Rudnitski, Anna
AU  - Kajaste-Rudnitski A
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Montini, Eugenio
AU  - Montini E
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy.
FAU - Penaud-Budloo, Magalie
AU  - Penaud-Budloo M
AD  - INSERM UMR 1089, University of Nantes, CHU of Nantes, Nantes 44200, France.
FAU - Naldini, Luigi
AU  - Naldini L
AD  - San Rafaelle Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific 
      Institute, Milan 20132, Italy; Vita-Salute San Raffaele University, Milan 20132, 
      Italy. Electronic address: naldini.luigi@hsr.it.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Cell Stem Cell
JT  - Cell stem cell
JID - 101311472
RN  - 0 (DNA, Viral)
RN  - 0 (Tumor Suppressor Protein p53)
RN  - EC 2.7.7.- (Integrases)
SB  - IM
CIN - Cell Stem Cell. 2022 Oct 6;29(10):1426-1427. PMID: 36206729
MH  - CRISPR-Cas Systems
MH  - DNA Damage
MH  - *DNA, Viral
MH  - Gene Editing
MH  - Hematopoietic Stem Cells
MH  - Humans
MH  - Integrases
MH  - *Tumor Suppressor Protein p53/genetics
PMC - PMC9550218
OTO - NOTNLM
OT  - CRISPR-Cas9
OT  - DNA damage response
OT  - DNA double-strand breaks
OT  - adeno-associated vector
OT  - gene editing
OT  - genotoxicity
OT  - hematopoietic stem/progenitor cells
OT  - integrase-defective lentiviral vector
OT  - inverted terminal repeats
OT  - p53 pathway
COIS- Declaration of interests L.N., A.K.-R., A.V., M.C.C., S.F., and A.J. are 
      inventors of patents on gene editing owned and managed by the San Raffaele 
      Scientific Institute and Telethon Foundation. L.N. is founder, quota holder, and 
      consultant of GeneSpire, a startup company aiming to develop ex vivo gene editing 
      and exploiting IDLV.
EDAT- 2022/10/08 06:00
MHDA- 2022/10/12 06:00
PMCR- 2022/10/06
CRDT- 2022/10/07 18:27
PHST- 2022/03/04 00:00 [received]
PHST- 2022/07/18 00:00 [revised]
PHST- 2022/09/06 00:00 [accepted]
PHST- 2022/10/08 06:00 [pubmed]
PHST- 2022/10/12 06:00 [medline]
PHST- 2022/10/07 18:27 [entrez]
PHST- 2022/10/06 00:00 [pmc-release]
AID - S1934-5909(22)00378-2 [pii]
AID - 10.1016/j.stem.2022.09.001 [doi]
PST - ppublish
SO  - Cell Stem Cell. 2022 Oct 6;29(10):1428-1444.e9. doi: 10.1016/j.stem.2022.09.001.