Isolating Malignant and Non-Malignant B Cells from &lt;em&gt;lck:eGFP&lt;/em&gt; Zebrafish

Burroughs‐Garcia, Jessica; Hasan, Ameera; Park, Gilseung; Borga, Chiara; Frazer, J. Kimble

doi:10.3791/59191

Cited by 9 publications

(12 citation statements)

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“…We discovered many D. rerio B lymphocytes and B-ALL express low levels of lck [58], which fostered new studies not previously done in zebrafish, such as FACS isolation of B-lineage cells (i. e., GFP lo cells from lck:GFP fish) and profiling their gene expression [58,63]. These findings translate to humans because immature human B cells and many patients' B-ALL also express low levels of LCK [58,75,76].…”

Section: Defining B-all and B Cell Gene Expression Patternsmentioning

confidence: 92%

“…Animals that developed fluorescent cancers were euthanized and single cell suspensions were prepared as described previously [58]. Cells in the lymphoid and precursor gates were analyzed for GFP intensity using a Beckman-Coulter CytoFLEX™ to determine whether each ALL derived from T, B, or both lymphocyte lineages [58,62,63].…”

Section: B-and T-all Incidence Studiesmentioning

confidence: 99%

“…ALL cells from double-transgenic hMYC fish were isolated from peritoneal washings as described previously [63]. Individual B-and T-ALL cells were FAC-sorted into 96 well plates, and cells were lysed, RNA extracted, and cDNA synthesized according to the Fluidigm™ single-cell protocol.…”

Section: Single-cell Qrt-pcr Analyses Of Hmyc B-and T-allmentioning

confidence: 99%

“…WT and hMYC GFP hi thymocytes express T cells genes, as do WT GFP hi marrow cells. WT GFP lo thymocyte fractions contain a mixture of B and T cells (as do hMYC GFP lo thymocytes and WT GFP lo marrow cells), due to the difficulty of obtaining pure GFP lo populations from these tissues [58,63]. Samples are clustered hierarchically.…”

Section: Distinct Types Of B-all Driven By Hmyc and Mmycmentioning

confidence: 99%

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Untitled

2020

Oncotarget

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Acute lymphoblastic leukemia (ALL) is the most common pediatric, and ninth most common adult, cancer. ALL can develop in either B or T lymphocytes, but B-lineage ALL (B-ALL) exceeds T-ALL clinically. As for other cancers, animal models allow study of the molecular mechanisms driving ALL. Several zebrafish (Danio rerio)T-ALL models have been reported, but until recently, robust D. rerio B-ALL models were not described. Then, D. rerio B-ALL was discovered in two related zebrafish transgenic lines; both were already known to develop T-ALL. Here, we report new B-ALL findings in one of these models, fish expressing transgenic human MYC (hMYC). We describe B-ALL incidence in a large cohort of hMYC fish, and show B-ALL in two new lines where T-ALL does not interfere with B-ALL detection. We also demonstrate B-ALL responses to steroid and radiation treatments, which effect ALL remissions, but are usually followed by prompt relapses. Finally, we report gene expression in zebrafish B lymphocytes and B-ALL, in both bulk samples and single B-and T-ALL cells. Using these gene expression profiles, we compare differences between the two new D. rerio B-ALL models, which are both driven by transgenic mammalian MYC oncoproteins. Collectively, these new data expand the utility of this new vertebrate B-ALL model.

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Section: Defining B-all and B Cell Gene Expression Patternsmentioning

confidence: 92%

Section: B-and T-all Incidence Studiesmentioning

confidence: 99%

Section: Single-cell Qrt-pcr Analyses Of Hmyc B-and T-allmentioning

confidence: 99%

Section: Distinct Types Of B-all Driven By Hmyc and Mmycmentioning

confidence: 99%

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2020

Oncotarget

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“…They built a double-transgenic with hMYC in addition to an lck : EGFP marker transgene. Fortuitously, this marker is highly expressed by T cells, but lowly expressed by some—but not all—B cells [90]. Consequently, in rag2:MYC-ER ; lck:EGFP fish , T-ALL are GFP hi , while B-ALL are GFP lo .…”

Section: Zebrafish Acute Lymphoblastic Leukemia Modelsmentioning

confidence: 99%

Tackling Acute Lymphoblastic Leukemia—One Fish at a Time

Sinha

Park

Frazer

2019

IJMS

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Despite advancements in the diagnosis and treatment of acute lymphoblastic leukemia (ALL), a need for improved strategies to decrease morbidity and improve cure rates in relapsed/refractory ALL still exists. Such approaches include the identification and implementation of novel targeted combination regimens, and more precise upfront patient risk stratification to guide therapy. New curative strategies rely on an understanding of the pathobiology that derives from systematically dissecting each cancer’s genetic and molecular landscape. Zebrafish models provide a powerful system to simulate human diseases, including leukemias and ALL specifically. They are also an invaluable tool for genetic manipulation, in vivo studies, and drug discovery. Here, we highlight and summarize contributions made by several zebrafish T-ALL models and newer zebrafish B-ALL models in translating the underlying genetic and molecular mechanisms operative in ALL, and also highlight their potential utility for drug discovery. These models have laid the groundwork for increasing our understanding of the molecular basis of ALL to further translational and clinical research endeavors that seek to improve outcomes in this important cancer.

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Dynamic Changes in Lymphocyte Populations Establish Zebrafish as a Thymic Involution Model

Hasan¹,

Macı́as²,

Wood³

et al. 2023

Preprint

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The thymus is the site of T lymphocyte development and T cell education to recognize foreign, but not self antigens. B cells also reside and develop in the thymus, although their functions are less clear. During thymic involution, a process of lymphoid atrophy and adipose replacement linked to sexual maturation, thymocytes decline. However, thymic B cells decrease far less than T cells, such that B cells comprise ~1% of human neonatal thymocytes, but up to ~10% in adults. All jawed vertebrates possess a thymus, and we and others have shown zebrafish (Danio rerio) also have thymic B cells. Here, we investigated the precise identities of zebrafish thymic T and B cells and how they change with involution. We assessed the timing and specific details of zebrafish thymic involution using multiple lymphocyte-specific, fluorophore-labeled transgenic lines, quantifying the changes in thymic T- and B-lymphocytes pre- vs. post-involution. Our results prove that, as in humans, zebrafish thymic B cells increase relative to T cells post-involution. We also performed RNA sequencing (RNA-seq) on D. rerio thymic and marrow lymphocytes of four novel double-transgenic lines, identifying distinct populations of immature T and B cells. Collectively, this is the first comprehensive analysis of zebrafish thymic involution, demonstrating its similarity to human involution, and establishing the highly genetically-manipulatable zebrafish model as a template for involution studies.

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Isolating Malignant and Non-Malignant B Cells from <em>lck:eGFP</em> Zebrafish

Cited by 9 publications

References 0 publications

Untitled

Untitled

Tackling Acute Lymphoblastic Leukemia—One Fish at a Time

Dynamic Changes in Lymphocyte Populations Establish Zebrafish as a Thymic Involution Model

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