Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy
In this work we provide preclinical data to support initiation of a first-in-human trial for sickle cell disease (SCD) using an approach that relies on reversal of the developmental fetal-to-adult hemoglobin switch. Erythroid-specific knockdown of BCL11A via a lentiviral-encoded microRNA-adapted sho...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050120300450 |
| _version_ | 1819076811632410624 |
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| author | Christian Brendel Olivier Negre Michael Rothe Swaroopa Guda Geoff Parsons Chad Harris Meaghan McGuinness Daniela Abriss Alla Tsytsykova Denise Klatt Martin Bentler Danilo Pellin Lauryn Christiansen Axel Schambach John Manis Helene Trebeden-Negre Melissa Bonner Erica Esrick Gabor Veres Myriam Armant David A. Williams |
| author_facet | Christian Brendel Olivier Negre Michael Rothe Swaroopa Guda Geoff Parsons Chad Harris Meaghan McGuinness Daniela Abriss Alla Tsytsykova Denise Klatt Martin Bentler Danilo Pellin Lauryn Christiansen Axel Schambach John Manis Helene Trebeden-Negre Melissa Bonner Erica Esrick Gabor Veres Myriam Armant David A. Williams |
| author_sort | Christian Brendel |
| collection | DOAJ |
| description | In this work we provide preclinical data to support initiation of a first-in-human trial for sickle cell disease (SCD) using an approach that relies on reversal of the developmental fetal-to-adult hemoglobin switch. Erythroid-specific knockdown of BCL11A via a lentiviral-encoded microRNA-adapted short hairpin RNA (shRNAmiR) leads to reactivation of the gamma-globin gene while simultaneously reducing expression of the pathogenic adult sickle β-globin. We generated a refined lentiviral vector (LVV) BCH-BB694 that was developed to overcome poor vector titers observed in the manufacturing scale-up of the original research-grade LVV. Healthy or sickle cell donor CD34+ cells transduced with Good Manufacturing Practices (GMP)-grade BCH-BB694 LVV achieved high vector copy numbers (VCNs) >5 and gene marking of >80%, resulting in a 3- to 5-fold induction of fetal hemoglobin (HbF) compared with mock-transduced cells without affecting growth, differentiation, and engraftment of gene-modified cells in vitro or in vivo. In vitro immortalization assays, which are designed to measure vector-mediated genotoxicity, showed no increased immortalization compared with mock-transduced cells. Together these data demonstrate that BCH-BB694 LVV is non-toxic and efficacious in preclinical studies, and can be generated at a clinically relevant scale in a GMP setting at high titer to support clinical testing for the treatment of SCD. |
| first_indexed | 2024-12-21T18:47:14Z |
| format | Article |
| id | doaj.art-7ebf28bc3d664f328d3cea76508fe211 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-21T18:47:14Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-7ebf28bc3d664f328d3cea76508fe2112022-12-21T18:53:51ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012020-06-0117589600Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene TherapyChristian Brendel0Olivier Negre1Michael Rothe2Swaroopa Guda3Geoff Parsons4Chad Harris5Meaghan McGuinness6Daniela Abriss7Alla Tsytsykova8Denise Klatt9Martin Bentler10Danilo Pellin11Lauryn Christiansen12Axel Schambach13John Manis14Helene Trebeden-Negre15Melissa Bonner16Erica Esrick17Gabor Veres18Myriam Armant19David A. Williams20Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA; Harvard Medical School, Boston, MA, USA; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USAbluebird bio, Inc., Cambridge, MA, USAInstitute of Experimental Hematology, Hannover Medical School, Hannover, GermanyDivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USAbluebird bio, Inc., Cambridge, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA; Institute of Experimental Hematology, Hannover Medical School, Hannover, GermanyDivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA; Institute of Experimental Hematology, Hannover Medical School, Hannover, GermanyDivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA; Harvard Medical School, Boston, MA, USA; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USAbluebird bio, Inc., Cambridge, MA, USAInstitute of Experimental Hematology, Hannover Medical School, Hannover, GermanyDepartment of Laboratory Medicine, Boston Children’s Hospital, Boston, MA, USAConnell & O’Reilly Families Cell Manipulation Core Facility, Dana-Farber Cancer Institute, Boston, MA, USAbluebird bio, Inc., Cambridge, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USAbluebird bio, Inc., Cambridge, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USADivision of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA; Harvard Medical School, Boston, MA, USA; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA; Corresponding author: David A. Williams, Boston Children’s Hospital, 300 Longwood Avenue, Karp 08125.3, Boston, MA 02115, USA.In this work we provide preclinical data to support initiation of a first-in-human trial for sickle cell disease (SCD) using an approach that relies on reversal of the developmental fetal-to-adult hemoglobin switch. Erythroid-specific knockdown of BCL11A via a lentiviral-encoded microRNA-adapted short hairpin RNA (shRNAmiR) leads to reactivation of the gamma-globin gene while simultaneously reducing expression of the pathogenic adult sickle β-globin. We generated a refined lentiviral vector (LVV) BCH-BB694 that was developed to overcome poor vector titers observed in the manufacturing scale-up of the original research-grade LVV. Healthy or sickle cell donor CD34+ cells transduced with Good Manufacturing Practices (GMP)-grade BCH-BB694 LVV achieved high vector copy numbers (VCNs) >5 and gene marking of >80%, resulting in a 3- to 5-fold induction of fetal hemoglobin (HbF) compared with mock-transduced cells without affecting growth, differentiation, and engraftment of gene-modified cells in vitro or in vivo. In vitro immortalization assays, which are designed to measure vector-mediated genotoxicity, showed no increased immortalization compared with mock-transduced cells. Together these data demonstrate that BCH-BB694 LVV is non-toxic and efficacious in preclinical studies, and can be generated at a clinically relevant scale in a GMP setting at high titer to support clinical testing for the treatment of SCD.http://www.sciencedirect.com/science/article/pii/S2329050120300450Sickle cell diseaseHbShemoglobinopathiesgene therapylentiviral vectorBCL11A |
| spellingShingle | Christian Brendel Olivier Negre Michael Rothe Swaroopa Guda Geoff Parsons Chad Harris Meaghan McGuinness Daniela Abriss Alla Tsytsykova Denise Klatt Martin Bentler Danilo Pellin Lauryn Christiansen Axel Schambach John Manis Helene Trebeden-Negre Melissa Bonner Erica Esrick Gabor Veres Myriam Armant David A. Williams Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy Molecular Therapy: Methods & Clinical Development Sickle cell disease HbS hemoglobinopathies gene therapy lentiviral vector BCL11A |
| title | Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy |
| title_full | Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy |
| title_fullStr | Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy |
| title_full_unstemmed | Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy |
| title_short | Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy |
| title_sort | preclinical evaluation of a novel lentiviral vector driving lineage specific bcl11a knockdown for sickle cell gene therapy |
| topic | Sickle cell disease HbS hemoglobinopathies gene therapy lentiviral vector BCL11A |
| url | http://www.sciencedirect.com/science/article/pii/S2329050120300450 |
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