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Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin
β-hemoglobinopathies are caused by abnormal or absent production of hemoglobin in the blood due to mutations in the β-globin gene (HBB). Imbalanced expression of adult hemoglobin (HbA) induces strong anemia in patients suffering from the disease. However, individuals with natural-occurring mutations...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7311455/ https://www.ncbi.nlm.nih.gov/pubmed/32576837 http://dx.doi.org/10.1038/s41598-020-66309-x |
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author | Lamsfus-Calle, Andrés Daniel-Moreno, Alberto Antony, Justin S. Epting, Thomas Heumos, Lukas Baskaran, Praveen Admard, Jakob Casadei, Nicolas Latifi, Ngadhnjim Siegmund, Darina M. Kormann, Michael S. D. Handgretinger, Rupert Mezger, Markus |
author_facet | Lamsfus-Calle, Andrés Daniel-Moreno, Alberto Antony, Justin S. Epting, Thomas Heumos, Lukas Baskaran, Praveen Admard, Jakob Casadei, Nicolas Latifi, Ngadhnjim Siegmund, Darina M. Kormann, Michael S. D. Handgretinger, Rupert Mezger, Markus |
author_sort | Lamsfus-Calle, Andrés |
collection | PubMed |
description | β-hemoglobinopathies are caused by abnormal or absent production of hemoglobin in the blood due to mutations in the β-globin gene (HBB). Imbalanced expression of adult hemoglobin (HbA) induces strong anemia in patients suffering from the disease. However, individuals with natural-occurring mutations in the HBB cluster or related genes, compensate this disparity through γ-globin expression and subsequent fetal hemoglobin (HbF) production. Several preclinical and clinical studies have been performed in order to induce HbF by knocking-down genes involved in HbF repression (KLF1 and BCL11A) or disrupting the binding sites of several transcription factors in the γ-globin gene (HBG1/2). In this study, we thoroughly compared the different CRISPR/Cas9 gene-disruption strategies by gene editing analysis and assessed their safety profile by RNA-seq and GUIDE-seq. All approaches reached therapeutic levels of HbF after gene editing and showed similar gene expression to the control sample, while no significant off-targets were detected by GUIDE-seq. Likewise, all three gene editing platforms were established in the GMP-grade CliniMACS Prodigy, achieving similar outcome to preclinical devices. Based on this gene editing comparative analysis, we concluded that BCL11A is the most clinically relevant approach while HBG1/2 could represent a promising alternative for the treatment of β-hemoglobinopathies. |
format | Online Article Text |
id | pubmed-7311455 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-73114552020-06-25 Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin Lamsfus-Calle, Andrés Daniel-Moreno, Alberto Antony, Justin S. Epting, Thomas Heumos, Lukas Baskaran, Praveen Admard, Jakob Casadei, Nicolas Latifi, Ngadhnjim Siegmund, Darina M. Kormann, Michael S. D. Handgretinger, Rupert Mezger, Markus Sci Rep Article β-hemoglobinopathies are caused by abnormal or absent production of hemoglobin in the blood due to mutations in the β-globin gene (HBB). Imbalanced expression of adult hemoglobin (HbA) induces strong anemia in patients suffering from the disease. However, individuals with natural-occurring mutations in the HBB cluster or related genes, compensate this disparity through γ-globin expression and subsequent fetal hemoglobin (HbF) production. Several preclinical and clinical studies have been performed in order to induce HbF by knocking-down genes involved in HbF repression (KLF1 and BCL11A) or disrupting the binding sites of several transcription factors in the γ-globin gene (HBG1/2). In this study, we thoroughly compared the different CRISPR/Cas9 gene-disruption strategies by gene editing analysis and assessed their safety profile by RNA-seq and GUIDE-seq. All approaches reached therapeutic levels of HbF after gene editing and showed similar gene expression to the control sample, while no significant off-targets were detected by GUIDE-seq. Likewise, all three gene editing platforms were established in the GMP-grade CliniMACS Prodigy, achieving similar outcome to preclinical devices. Based on this gene editing comparative analysis, we concluded that BCL11A is the most clinically relevant approach while HBG1/2 could represent a promising alternative for the treatment of β-hemoglobinopathies. Nature Publishing Group UK 2020-06-23 /pmc/articles/PMC7311455/ /pubmed/32576837 http://dx.doi.org/10.1038/s41598-020-66309-x Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Lamsfus-Calle, Andrés Daniel-Moreno, Alberto Antony, Justin S. Epting, Thomas Heumos, Lukas Baskaran, Praveen Admard, Jakob Casadei, Nicolas Latifi, Ngadhnjim Siegmund, Darina M. Kormann, Michael S. D. Handgretinger, Rupert Mezger, Markus Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin |
title | Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin |
title_full | Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin |
title_fullStr | Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin |
title_full_unstemmed | Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin |
title_short | Comparative targeting analysis of KLF1, BCL11A, and HBG1/2 in CD34(+) HSPCs by CRISPR/Cas9 for the induction of fetal hemoglobin |
title_sort | comparative targeting analysis of klf1, bcl11a, and hbg1/2 in cd34(+) hspcs by crispr/cas9 for the induction of fetal hemoglobin |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7311455/ https://www.ncbi.nlm.nih.gov/pubmed/32576837 http://dx.doi.org/10.1038/s41598-020-66309-x |
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