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Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency
Pyruvate kinase deficiency (PKD), an autosomal-recessive disorder, is the main cause of chronic non-spherocytic hemolytic anemia. PKD is caused by mutations in the pyruvate kinase, liver and red blood cell (PKLR) gene, which encodes for the erythroid pyruvate kinase protein (RPK). RPK is implicated...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399088/ https://www.ncbi.nlm.nih.gov/pubmed/34485608 http://dx.doi.org/10.1016/j.omtm.2021.05.001 |
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author | Fañanas-Baquero, Sara Quintana-Bustamante, Oscar Dever, Daniel P. Alberquilla, Omaira Sanchez-Dominguez, Rebeca Camarena, Joab Ojeda-Perez, Isabel Dessy-Rodriguez, Mercedes Turk, Rolf Schubert, Mollie S. Lattanzi, Annalisa Xu, Liwen Lopez-Lorenzo, Jose L. Bianchi, Paola Bueren, Juan A. Behlke, Mark A. Porteus, Matthew Segovia, Jose-Carlos |
author_facet | Fañanas-Baquero, Sara Quintana-Bustamante, Oscar Dever, Daniel P. Alberquilla, Omaira Sanchez-Dominguez, Rebeca Camarena, Joab Ojeda-Perez, Isabel Dessy-Rodriguez, Mercedes Turk, Rolf Schubert, Mollie S. Lattanzi, Annalisa Xu, Liwen Lopez-Lorenzo, Jose L. Bianchi, Paola Bueren, Juan A. Behlke, Mark A. Porteus, Matthew Segovia, Jose-Carlos |
author_sort | Fañanas-Baquero, Sara |
collection | PubMed |
description | Pyruvate kinase deficiency (PKD), an autosomal-recessive disorder, is the main cause of chronic non-spherocytic hemolytic anemia. PKD is caused by mutations in the pyruvate kinase, liver and red blood cell (PKLR) gene, which encodes for the erythroid pyruvate kinase protein (RPK). RPK is implicated in the last step of anaerobic glycolysis in red blood cells (RBCs), responsible for the maintenance of normal erythrocyte ATP levels. The only curative treatment for PKD is allogeneic hematopoietic stem and progenitor cell (HSPC) transplant, associated with a significant morbidity and mortality, especially relevant in PKD patients. Here, we address the correction of PKD through precise gene editing at the PKLR endogenous locus to keep the tight regulation of RPK enzyme during erythropoiesis. We combined CRISPR-Cas9 system and donor recombinant adeno-associated vector (rAAV) delivery to build an efficient, safe, and clinically applicable system to knock in therapeutic sequences at the translation start site of the RPK isoform in human hematopoietic progenitors. Edited human hematopoietic progenitors efficiently reconstituted human hematopoiesis in primary and secondary immunodeficient mice. Erythroid cells derived from edited PKD-HSPCs recovered normal ATP levels, demonstrating the restoration of RPK function in PKD erythropoiesis after gene editing. Our gene-editing strategy may represent a lifelong therapy to correct RPK functionality in RBCs for PKD patients. |
format | Online Article Text |
id | pubmed-8399088 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-83990882021-09-03 Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency Fañanas-Baquero, Sara Quintana-Bustamante, Oscar Dever, Daniel P. Alberquilla, Omaira Sanchez-Dominguez, Rebeca Camarena, Joab Ojeda-Perez, Isabel Dessy-Rodriguez, Mercedes Turk, Rolf Schubert, Mollie S. Lattanzi, Annalisa Xu, Liwen Lopez-Lorenzo, Jose L. Bianchi, Paola Bueren, Juan A. Behlke, Mark A. Porteus, Matthew Segovia, Jose-Carlos Mol Ther Methods Clin Dev Original Article Pyruvate kinase deficiency (PKD), an autosomal-recessive disorder, is the main cause of chronic non-spherocytic hemolytic anemia. PKD is caused by mutations in the pyruvate kinase, liver and red blood cell (PKLR) gene, which encodes for the erythroid pyruvate kinase protein (RPK). RPK is implicated in the last step of anaerobic glycolysis in red blood cells (RBCs), responsible for the maintenance of normal erythrocyte ATP levels. The only curative treatment for PKD is allogeneic hematopoietic stem and progenitor cell (HSPC) transplant, associated with a significant morbidity and mortality, especially relevant in PKD patients. Here, we address the correction of PKD through precise gene editing at the PKLR endogenous locus to keep the tight regulation of RPK enzyme during erythropoiesis. We combined CRISPR-Cas9 system and donor recombinant adeno-associated vector (rAAV) delivery to build an efficient, safe, and clinically applicable system to knock in therapeutic sequences at the translation start site of the RPK isoform in human hematopoietic progenitors. Edited human hematopoietic progenitors efficiently reconstituted human hematopoiesis in primary and secondary immunodeficient mice. Erythroid cells derived from edited PKD-HSPCs recovered normal ATP levels, demonstrating the restoration of RPK function in PKD erythropoiesis after gene editing. Our gene-editing strategy may represent a lifelong therapy to correct RPK functionality in RBCs for PKD patients. American Society of Gene & Cell Therapy 2021-05-14 /pmc/articles/PMC8399088/ /pubmed/34485608 http://dx.doi.org/10.1016/j.omtm.2021.05.001 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Fañanas-Baquero, Sara Quintana-Bustamante, Oscar Dever, Daniel P. Alberquilla, Omaira Sanchez-Dominguez, Rebeca Camarena, Joab Ojeda-Perez, Isabel Dessy-Rodriguez, Mercedes Turk, Rolf Schubert, Mollie S. Lattanzi, Annalisa Xu, Liwen Lopez-Lorenzo, Jose L. Bianchi, Paola Bueren, Juan A. Behlke, Mark A. Porteus, Matthew Segovia, Jose-Carlos Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency |
title | Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency |
title_full | Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency |
title_fullStr | Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency |
title_full_unstemmed | Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency |
title_short | Clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency |
title_sort | clinically relevant gene editing in hematopoietic stem cells for the treatment of pyruvate kinase deficiency |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399088/ https://www.ncbi.nlm.nih.gov/pubmed/34485608 http://dx.doi.org/10.1016/j.omtm.2021.05.001 |
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