Human cellular model systems of β-thalassemia enable in-depth analysis of disease phenotype

Debbie Daniels, Ivan Ferrer Vicens, Joseph S Hawksworth, Tatyana N Andrienko, Elizabeth F Finnie, Natalie S Bretherton, Daniel C J Ferguson, A Oliveira, Jenn-Yeu Szeto, Marieangela C Wilson, John N. Brewin, Jan Frayne*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

1 Citation (Scopus)

Abstract

β-thalassemia is a prevalent genetic disorder causing severe anemia due to defective erythropoiesis, with few treatment options. Studying the underlying molecular defects is impeded by paucity of suitable patient material. In this study we create human disease cellular model systems for β-thalassemia by gene editing the erythroid line BEL-A, which accurately recapitulate the phenotype of patient erythroid cells. We also develop a high throughput compatible fluorometric-based assay for evaluating severity of disease phenotype and utilize the assay to demonstrate that the lines respond appropriately to verified reagents. We next use the lines to perform extensive analysis of the altered molecular mechanisms in β-thalassemia erythroid cells, revealing upregulation of a wide range of biological pathways and processes along with potential novel targets for therapeutic investigation. Overall, the lines provide a sustainable supply of disease cells as research tools for identifying therapeutic targets and as screening platforms for new drugs and reagents.
Original languageEnglish
Article number6260
JournalNature Communications
Volume14
DOIs
Publication statusPublished - 6 Oct 2023

Bibliographical note

Funding Information:
The authors would like to thank Dr. Kate Heesom, Director of the University of Bristol Proteomics Facility, for performing mass spectrometry and analysis of raw data, Dr. Phil Lewis, School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK for mass spectrometry data analysis, the University of Bristol Flow Cytometry Facility for use of equipment, and Dr. Andrew Herman, director of, and Helen Rice of the Flow Cytometry Facility for performing cell sorting. The gRNAs used to generate the HBB−/−HBD−/−cell line were a gift from Professor Jacob Corn, ETH Zurich, Switzerland. We also thank the patients for their valuable participation in this study. The study was supported by MRC grant MR/S021140/1, Wellcome Trust grant 209739/Z/17/Z, the Global Challenges Research Fund via the University of Bristol’s GCRF and Newton Fund Consolidation Account (GNCA) and BBSRC Pathfinder award BB/X511195/1 via the University of Bristol. The work of J.A.S. was supported by BBSRC-funded SWBio DTP. A.S.F.O. acknowledges the BBSRC grant BB/X009831/1 and ERC (Advanced Grant PREDACTED https://cordis.europa.eu/project/id/101021207) for support. ASFO also thanks Oracle for Research for the Research Fellowship.

Funding Information:
The authors would like to thank Dr. Kate Heesom, Director of the University of Bristol Proteomics Facility, for performing mass spectrometry and analysis of raw data, Dr. Phil Lewis, School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK for mass spectrometry data analysis, the University of Bristol Flow Cytometry Facility for use of equipment, and Dr. Andrew Herman, director of, and Helen Rice of the Flow Cytometry Facility for performing cell sorting. The gRNAs used to generate the HBBHBD cell line were a gift from Professor Jacob Corn, ETH Zurich, Switzerland. We also thank the patients for their valuable participation in this study. The study was supported by MRC grant MR/S021140/1, Wellcome Trust grant 209739/Z/17/Z, the Global Challenges Research Fund via the University of Bristol’s GCRF and Newton Fund Consolidation Account (GNCA) and BBSRC Pathfinder award BB/X511195/1 via the University of Bristol. The work of J.A.S. was supported by BBSRC-funded SWBio DTP. A.S.F.O. acknowledges the BBSRC grant BB/X009831/1 and ERC (Advanced Grant PREDACTED https://cordis.europa.eu/project/id/101021207 ) for support. ASFO also thanks Oracle for Research for the Research Fellowship. −/− −/−

Publisher Copyright:
© 2023, Springer Nature Limited.

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