Establishment of a congenital amegakaryocytic thrombocytopenia model and a thrombocyte–specific reporter line in zebrafish

• Published in: Leukemia 2017-05, Vol.31 (5), p.1206-1216
• Main Authors: Lin, Q, Zhang, Y, Zhou, R, Zheng, Y, Zhao, L, Huang, M, Zhang, X, Leung, A Y H, Zhang, W
• Format: Article
• Language: English
• Subjects: 14/19; 38/23; 38/32; 45/77; 45/90; 631/80/86; 64/116; 96/21; 96/31; Analysis; Animals; Blood Platelets - pathology; Bone marrow; Bone marrow transplantation; Cancer Research; Congenital diseases; Critical Care Medicine; Danio rerio; Developmental biology; Disease Models, Animal; Drug screening; Fluorescence; Gene mutation; Genes, Reporter; Genetic aspects; Genetic engineering; Green fluorescent protein; Green Fluorescent Proteins - genetics; Hematology; Hematopoietic stem cells; Higher education; Humans; Intensive; Internal Medicine; Janus kinase 2; Janus Kinase 2 - metabolism; Leukemia; Medical prognosis; Medicine; Medicine & Public Health; Mimicry; Mutants; Mutation; Nuclease; Oncology; original-article; Pathogenesis; Physiological aspects; Platelets; Progenitor cells; Prognosis; Proteins; Receptors, Thrombopoietin - genetics; Receptors, Thrombopoietin - metabolism; Risk factors; Signal Transduction; Stem cell transplantation; Thrombocytes; Thrombocytopenia; Thrombocytopenia - pathology; Thrombopoietin - metabolism; Transcription; Transcription activator-like effector nucleases; Transplantation; Zebrafish
• ISSN: 0887-6924; 1476-5551
• DOI: 10.1038/leu.2016.320

Mutations in the human myeloproliferative leukemia ( MPL ) protein gene are known to cause congenital amegakaryocytic thrombocytopenia (CAMT). The prognosis of this heritable disorder is poor and bone marrow transplantation is the only effective treatment. Here, by using the TALEN (transcription activator-like effector nuclease) technology, we created a zebrafish mpl mutant to model human CAMT. Disruption of zebrafish mpl lead to a severe reduction in thrombocytes and a high bleeding tendency, as well as deficiencies in adult hematopoietic stem/progenitor cells. We further demonstrated that thrombocytopenia in mpl mutant zebrafish was caused by impaired Tpo/Mpl/Jak2 signaling, resulting in reduced proliferation of thrombocyte precursors. These results indicate that mpl mutant zebrafish develop thrombocytopenia resembling the human CAMT. To utilize fully zebrafish to study thrombocyte biology and thrombocytopenia disorders, we generated a transgenic reporter line Tg(mpl:eGFP)smu4 , in which green fluorescent protein (GFP) expression was driven by the mpl promoter. Detailed characterization of Tg(mpl:eGFP)smu4 fish confirmed that the thrombocyte lineage was specifically marked by GFP expression. In conclusion, we generated the first transmissible congenital thrombocytopenia zebrafish model mimicking human CAMT and a thrombocyte-specific transgenic line. Together with Tg(mpl:eGFP)smu4 , mpl mutant zebrafish provide a useful tool for drug screening and study of thrombocytopoiesis.