PUBLICATION
Generation of a thrombopoietin-deficient thrombocytopenia model in zebrafish
- Authors
- Yang, L., Wu, L., Meng, P., Zhang, X., Zhao, D., Lin, Q., Zhang, Y.
- ID
- ZDB-PUB-220528-5
- Date
- 2022
- Source
- Journal of thrombosis and haemostasis : JTH 20(8): 1900-1909 (Journal)
- Registered Authors
- Lin, Qing, Zhang, Yiyue
- Keywords
- Disease model, Human clinical variants, Thpo, Thrombocytopenia, zebrafish
- MeSH Terms
-
- Animals
- Disease Models, Animal
- Humans
- Receptors, Thrombopoietin/genetics
- Receptors, Thrombopoietin/metabolism
- Thrombocytopenia*/genetics
- Thrombocytopenia*/metabolism
- Thrombopoiesis/genetics
- Thrombopoietin*/genetics
- Thrombopoietin*/metabolism
- Zebrafish/genetics
- Zebrafish/metabolism
- PubMed
- 35622056 Full text @ J. Thromb. Haemost.
Citation
Yang, L., Wu, L., Meng, P., Zhang, X., Zhao, D., Lin, Q., Zhang, Y. (2022) Generation of a thrombopoietin-deficient thrombocytopenia model in zebrafish. Journal of thrombosis and haemostasis : JTH. 20(8):1900-1909.
Abstract
Background The production of platelets is tightly regulated by thrombopoietin (THPO). Mutations in the THPO gene cause thrombocytopenia. Although mice lacking Thpo present with thrombocytopenia, predicting phenotypes and pathogenicity of novel THPO mutations in mice is limited. Zebrafish can be a powerful tool for fast validation and study of candidate genes of human hematological diseases and have already been used as a model of human thrombocytopenia.
Objectives We aim to investigate the role of Thpo in zebrafish thrombopoiesis and to establish a Thpo-deficient zebrafish model. The model could be applied for illustrating the clinically discovered human THPO variants of which the clinical significance is not known and to evaluate the effect of THPO-RAs, as well as a screening platform for new drugs.
Methods We generated a thpo loss-of-function zebrafish model using CRISPR/Cas9. After disruption of zebrafish thpo, thposzy6 zebrafish presented with a significant reduction of thpo expression and developed thrombocytopenia. Furthermore, we performed in vivo studies with zebrafish with the thposzy6 mutation and found two human clinical point-mutations (c.091C>T and c.112C>T) that were responsible for the thrombocytopenia phenotype. In addition, effects of THPO receptor agonists (THPO-RAs) used as therapeutics against thrombocytopenia were evaluated in the Tg(mpl:eGFP);thposzy6 line.
Results and conclusions Zebrafish with the mutation thposzy6 presented with a significant reduction of thpo expression and developed thrombocytopenia. thpo loss-of-function zebrafish model can serve as a valuable preclinical model for thrombocytopenia caused by thpo-deficiency, as well as a tool to study human clinical THPO variants and evaluate the effect of THPO-RAs.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping