Generation and Characterization of a Zebrafish IL-2Rγc SCID Model

Sertori, Robert; Jones, Realla; Basheer, Faiza; Rivera, Leni R.; Dawson, Steve; Loke, Stella; Heidary, Somayyeh; Dhillon, Amardeep S.; Liongue, Clifford; Ward, Alister C.

doi:10.3390/ijms23042385

Cited by 15 publications

(26 citation statements)

References 47 publications

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“…JAK3 is exclusively associated with the common cytokine receptor chain IL-2Rγc [ 18 ]. Zebrafish possesses two paralogues for the human IL2RG gene, with il2rga shown to have a conserved role in lymphopoiesis acting upstream of jak3 [ 59 , 60 ]. Therefore, to elucidate the role of IL-2Rγc in mediating the effects of Jak3 A573V, fish carrying jak3 ca alleles were crossed with those carrying knockout alleles for il2rga [ 59 ] to produce il2rga +/− jak3 + /ca fish that were in-crossed to generate progeny that were assessed for rag1 expression and genotyped in parallel (Supp.…”

Section: Resultsmentioning

confidence: 99%

“…Carriers of the jak3 mutant allele were identified and, following an additional round of outcrossing, were in-crossed to generate wild-type, heterozygote and homozygote mutant progeny for analysis. The jak3 mutant was also crossed onto lines carrying hypomorphic mutations in il2rga [ 59 ], as well as jak1 , stat5.1 and stat5.2 produced in-house (unpublished data) to generate additional genotypes for analysis.…”

Section: Methodsmentioning

confidence: 99%

“…Zebrafish show strong conservation of cytokine receptor signaling through the JAK-STAT pathway [ 37 , 38 ], including a conserved role for the IL-2Rγc in immune cell development [ 59 , 60 ]. This study examined the A572V and A573V JAK3 PKD mutations, which were introduced into zebrafish Jak3 and the effects on STAT5 activation in vitro and T lymphocyte production in vivo analyzed.…”

Section: Introductionmentioning

confidence: 99%

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In vivo impact of JAK3 A573V mutation revealed using zebrafish

Basheer

Bulleeraz

Ngo

et al. 2022

Cell. Mol. Life Sci.

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Background Janus kinase 3 (JAK3) acts downstream of the interleukin-2 (IL-2) receptor family to play a pivotal role in the regulation of lymphoid cell development. Activating JAK3 mutations are associated with a number of lymphoid and other malignancies, with mutations within the regulatory pseudokinase domain common. Methods The pseudokinase domain mutations A572V and A573V were separately introduced into the highly conserved zebrafish Jak3 and transiently expressed in cell lines and zebrafish embryos to examine their activity and impact on early T cells. Genome editing was subsequently used to introduce the A573V mutation into the zebrafish genome to study the effects of JAK3 activation on lymphoid cells in a physiologically relevant context throughout the life-course. Results Zebrafish Jak3 A573V produced the strongest activation of downstream STAT5 in vitro and elicited a significant increase in T cells in zebrafish embryos. Zebrafish carrying just a single copy of the Jak3 A573V allele displayed elevated embryonic T cells, which continued into adulthood. Hematopoietic precursors and NK cells were also increased, but not B cells. The lymphoproliferative effects of Jak3 A573V in embryos was shown to be dependent on zebrafish IL-2Rγc, JAK1 and STAT5B equivalents, and could be suppressed with the JAK3 inhibitor Tofacitinib. Conclusions This study demonstrates that a single JAK3 A573V allele expressed from the endogenous locus was able to enhance lymphopoiesis throughout the life-course, which was mediated via an IL-2Rγc/JAK1/JAK3/STAT5 signaling pathway and was sensitive to Tofacitinib. This extends our understanding of oncogenic JAK3 mutations and creates a novel model to underpin further translational investigations.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In vivo impact of JAK3 A573V mutation revealed using zebrafish

Basheer

Bulleeraz

Ngo

et al. 2022

Cell. Mol. Life Sci.

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“…Based on this method, a knockout of gas7 promoting neuroblastoma metastasis in transgenic fish overexpressing MYCN was obtained [ 69 ]. Recently, TALEN has been used for the generation of immune-deficient models by targeting interleukin-2 receptor gamma common (IL-2Rγc) [ 70 ] or DNA-dependent protein kinase (Prkdc) [ 39 ].…”

Section: Disease Induction: How To Create a Fish Modelmentioning

confidence: 99%

One Host-Multiple Applications: Zebrafish (Danio rerio) as Promising Model for Studying Human Cancers and Pathogenic Diseases

Dudziak

Nowak

Sozoniuk

2022

IJMS

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In recent years, zebrafish (ZF) has been increasingly applied as a model in human disease studies, with a particular focus on cancer. A number of advantages make it an attractive alternative for mice widely used so far. Due to the many advantages of zebrafish, modifications can be based on different mechanisms and the induction of human disease can take different forms depending on the research goal. Genetic manipulation, tumor transplantation, or injection of the pathogen are only a few examples of using ZF as a model. Most of the studies are conducted in order to understand the disease mechanism, monitor disease progression, test new or alternative therapies, and select the best treatment. The transplantation of cancer cells derived from patients enables the development of personalized medicine. To better mimic a patient’s body environment, immune-deficient models (SCID) have been developed. A lower immune response is mostly generated by genetic manipulation but also by irradiation or dexamethasone treatment. For many studies, using SCID provides a better chance to avoid cancer cell rejection. In this review, we describe the main directions of using ZF in research, explain why and how zebrafish can be used as a model, what kind of limitations will be met and how to overcome them. We collected recent achievements in this field, indicating promising perspectives for the future.

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“…Zebrafish B lymphocytes develop around 20 dpf ( 18 ), with NK-related cells evident at this time ( 20 ). Conserved cytokine receptor homologues have been shown to be expressed in and contribute to the development of blood and immune cells, including erythropoietin receptor (EPO-R) for erythrocytes ( 21 ), granulocyte colony-stimulating factor receptor (GCSF-R) for granulocytes ( 22 ) and interleukin 2 receptor (IL-2R) family members for T lymphocytes ( 23 ).…”

Section: Introductionmentioning

confidence: 99%

Cytokine Receptor-Like Factor 3 (CRLF3) Contributes to Early Zebrafish Hematopoiesis

et al. 2022

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Cytokine receptor-like factor 3 (CRLF3) is an ancient protein conserved across metazoans that contains an archetypal cytokine receptor homology domain (CHD). This domain is found in cytokine receptors present in bilateria, including higher vertebrates, that play key roles in a variety of developmental and homeostatic processes, particularly relating to blood and immune cells. However, understanding of CRLF3 itself remains very limited. This study aimed to investigate this evolutionarily significant protein by studying its embryonic expression and function in early development, particularly of blood and immune cells, using zebrafish as a model. Expression of crlf3 was identified in mesoderm-derived tissues in early zebrafish embryos, including the somitic mesoderm and both anterior and posterior lateral plate mesoderm. Later expression was observed in the thymus, brain, retina and exocrine pancreas. Zebrafish crlf3 mutants generated by genome editing technology exhibited a significant reduction in primitive hematopoiesis and early definitive hematopoiesis, with decreased early progenitors impacting on multiple lineages. No other obvious phenotypes were observed in the crlf3 mutants.

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Generation and Characterization of a Zebrafish IL-2Rγc SCID Model

Cited by 15 publications

References 47 publications

In vivo impact of JAK3 A573V mutation revealed using zebrafish

In vivo impact of JAK3 A573V mutation revealed using zebrafish

One Host-Multiple Applications: Zebrafish (Danio rerio) as Promising Model for Studying Human Cancers and Pathogenic Diseases

Cytokine Receptor-Like Factor 3 (CRLF3) Contributes to Early Zebrafish Hematopoiesis

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