Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Engineered human Diamond-Blackfan anemia disease model confirms therapeutic effects of clinically applicable lentiviral vector at single-cell resolution

Liu, Yang LU ; Schmiderer, Ludwig LU ; Hjort, Martin LU orcid ; Lang, Stefan LU orcid ; Bremborg, Tyra LU orcid ; Rydström, Anna LU ; Schambach, Axel ; Larsson, Jonas LU and Karlsson, Stefan LU (2023) In Haematologica 108(11). p.3095-3109
Abstract

Diamond-Blackfan anemia is a rare genetic bone marrow failure disorder which is usually caused by mutations in ribosomal protein genes. In the present study, we generated a traceable RPS19-deficient cell model using CRISPR-Cas9 and homology-directed repair to investigate the therapeutic effects of a clinically applicable lentiviral vector at single-cell resolution. We developed a gentle nanostraw delivery platform to edit RPS19 gene in primary human cord blood-derived CD34+ hematopoietic stem and progenitor cells. The edited cells showed expected impaired erythroid differentiation phenotype and a specific erythroid progenitor with abnormal cell cycle status accompanied by enrichment of TNFα/NF-κB and p53 signaling pathways was... (More)

Diamond-Blackfan anemia is a rare genetic bone marrow failure disorder which is usually caused by mutations in ribosomal protein genes. In the present study, we generated a traceable RPS19-deficient cell model using CRISPR-Cas9 and homology-directed repair to investigate the therapeutic effects of a clinically applicable lentiviral vector at single-cell resolution. We developed a gentle nanostraw delivery platform to edit RPS19 gene in primary human cord blood-derived CD34+ hematopoietic stem and progenitor cells. The edited cells showed expected impaired erythroid differentiation phenotype and a specific erythroid progenitor with abnormal cell cycle status accompanied by enrichment of TNFα/NF-κB and p53 signaling pathways was identified by single-cell RNA sequencing analysis. The therapeutic vector could rescue the abnormal erythropoiesis by activating cell cycle-related signaling pathways and promoted red blood cell production. Overall, these results establish nanostraws as a gentle option for CRISPR-Cas9-based gene editing in sensitive primary hematopoietic stem and progenitor cells, and provide support for future clinical investigations of the lentiviral gene therapy strategy.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Haematologica
volume
108
issue
11
pages
47 pages
publisher
Ferrata Storti Foundation
external identifiers
  • scopus:85176262088
  • pmid:37199130
ISSN
1592-8721
DOI
10.3324/haematol.2022.282068
language
English
LU publication?
yes
id
e4c7100d-7fdf-4704-9fa9-e0233a002494
date added to LUP
2023-10-24 15:24:59
date last changed
2024-04-18 13:04:30
@article{e4c7100d-7fdf-4704-9fa9-e0233a002494,
  abstract     = {{<p>Diamond-Blackfan anemia is a rare genetic bone marrow failure disorder which is usually caused by mutations in ribosomal protein genes. In the present study, we generated a traceable RPS19-deficient cell model using CRISPR-Cas9 and homology-directed repair to investigate the therapeutic effects of a clinically applicable lentiviral vector at single-cell resolution. We developed a gentle nanostraw delivery platform to edit RPS19 gene in primary human cord blood-derived CD34+ hematopoietic stem and progenitor cells. The edited cells showed expected impaired erythroid differentiation phenotype and a specific erythroid progenitor with abnormal cell cycle status accompanied by enrichment of TNFα/NF-κB and p53 signaling pathways was identified by single-cell RNA sequencing analysis. The therapeutic vector could rescue the abnormal erythropoiesis by activating cell cycle-related signaling pathways and promoted red blood cell production. Overall, these results establish nanostraws as a gentle option for CRISPR-Cas9-based gene editing in sensitive primary hematopoietic stem and progenitor cells, and provide support for future clinical investigations of the lentiviral gene therapy strategy.</p>}},
  author       = {{Liu, Yang and Schmiderer, Ludwig and Hjort, Martin and Lang, Stefan and Bremborg, Tyra and Rydström, Anna and Schambach, Axel and Larsson, Jonas and Karlsson, Stefan}},
  issn         = {{1592-8721}},
  language     = {{eng}},
  number       = {{11}},
  pages        = {{3095--3109}},
  publisher    = {{Ferrata Storti Foundation}},
  series       = {{Haematologica}},
  title        = {{Engineered human Diamond-Blackfan anemia disease model confirms therapeutic effects of clinically applicable lentiviral vector at single-cell resolution}},
  url          = {{http://dx.doi.org/10.3324/haematol.2022.282068}},
  doi          = {{10.3324/haematol.2022.282068}},
  volume       = {{108}},
  year         = {{2023}},
}