Zebrafish Hematological Disease Models

Hematopoiesis is a complex process with various signaling pathways influencing every step of blood cell formation from the earliest precursors to final differentiated blood cell types. The formation of blood cells is vital for survival. Hematopoietic Stem and Progenitor Cells (HSPCs) are responsible for creating all adult differentiated blood cells. Genes that are mutated in hematopoietic malignancies and disease are often required for the normal maintenance and production of blood cells not only in adult tissues but also in the developing embryo. Defects in HSPCs or their downstream lineages can result in anemia and other hematological disorders including leukemia. Thus, understanding the molecular and cellular mechanisms that drive normal hematopoiesis and hematopoietic malignancies is critical for developing new treatments for blood diseases.

Zebrafish are being increasingly used as a model organism to investigate vertebrate hematopoiesis. This genetically tractable model has a number of appealing features, such as the ease of manipulation of transparent embryos and the capacity to conduct large-scale genetic and chemical screens. Zebrafish shares, with other vertebrates, all major blood cell types that are generated by similar developmental pathways. Besides, many of the genes known to regulate erythropoiesis in mammals have been identified in zebrafish. There is also a high conservation of transcription factors and major signaling pathways involved in hematopoietic stem cell (HSC) maturation and differentiation. As in mammals, zebrafish blood development contains three sequential waves: the primitive one, the erythromyeloid progenitor (EPM-derived) wave and the definitive wave that occurs in different embryonic locations. Thus, the parallels between the hematopoietic system in zebrafish and humans prompt the increasing use of zebrafish to model hematological disorders.

Figure 1. Zebrafish hematopoiesis and its key regulators. (Rasighaemi P, et al. 2015)

Zebrafish Models of Hematological Disease

Creative Biogene, a zebrafish preclinical contract research organization, has modelled a significant number of human hematopoietic malignancies in zebrafish by genome editing technologies such as TALENs and CRISPR/Cas9 systems. The models were developed by generating stable transgenic lines or through transient overexpression of human or zebrafish orthologs of oncogenes associated with hematopoietic malignancies. In addition, we can also use cell-type specific promoters to drive the expression of human oncogenes and fusion gene cDNAs in zebrafish.

Human Disease Condition	Zebrafish Transgenic Line	Affected Gene
Hereditary elliptocytosis	chablis and merlot	epb41b
Hereditary spherocytosis type 2	riesling	b-spectrin
Hereditary spherocytosis type 4	retsina	slc4a1
Erythropoietic protoporphyria and X-linked sideroblastic anemia	sauternes	alas2
Hypochromic microcytic anemia	zinfandel	hbbe1.1
Hypochromic microcytic anemia	chianti	tfr1a
Hemochromatosis type 4	weissherbst	slc40a1
Erythropoietic protoporphyria	dracula	fech
Diamond-Blackfan anemia	rpl11	rpl11
Acute myeloid leukemia	Tg(hsp:AML1-ETO)	runx1
Acute myeloid leukemia	Tg(spi1:lox-EGFP-loxNUP98-HOXA9)	hoxa9
Myelodysplastic syndrome	Tet2 mutants	Tet2
Myelodysplastic syndrome	crimsonless	hspa9b
T-cell acute lymphoblastic leukemia	Tg(rag2:EGFP-myc)	Myc-oncogene overexpression
T-cell acute lymphoblastic leukemia	Tg(rag2:NOTCH1ICD-EGFP)	Notch1-intracellular domain overexpression
Pre-B cell acute lymphoblastic leukemia	Tg(ef1alpha:EGFP-TELAML1)	runx1

Advantages

Small size, high fecundity, and fast embryonic development make zebrafish suitable for large-scale forward and modifier genetic screens.
Phenotype-based screens provide unbiased identification of candidate genetic pathways.
Zebrafish blood disease models have the potential to reveal new drugs for human disease treatment.
The transparency of zebrafish embryos allows live imaging of hematopoietic cells and of the pathologies of the diseases that affect them.

With extensive experience in zebrafish research, our scientists can help you choose the right model and experimental design to achieve your research and development goals.

References

Zizioli D, et al. Zebrafish disease models in hematology: highlights on biological and translational impact. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, 2019, 1865(3): 620-633.
Rissone A, Burgess S M. Rare genetic blood disease modeling in zebrafish. Frontiers in genetics, 2018, 9: 348.
Gore A V, et al. The zebrafish: A fintastic model for hematopoietic development and disease. Wiley Interdisciplinary Reviews: Developmental Biology, 2018, 7(3): e312.
Jing L, Zon L I. Zebrafish as a model for normal and malignant hematopoiesis. Disease models & mechanisms, 2011, 4(4): 433-438.
Rasighaemi P, et al. Zebrafish as a model for leukemia and other hematopoietic disorders. Journal of hematology & oncology, 2015, 8(1): 1-10.

For research use only. Not intended for any clinical use.