Long-term drug administration in the adult zebrafish using oral gavage for cancer preclinical studies.
Disease Models & Mechanisms 9(7):811 (2016)
Zebrafish are a major model for chemical genetics, and most studies use embryos when investigating small molecules that cause interesting phenotypes or that can rescue disease models. Limited studies have dosed adults with small molecules by means of water-borne exposure or injection techniques....
Engineering Hematopoietic Stem Cells: Lessons from Development.
Cell Stem Cell 18(6):707 (2016)
Cell engineering has brought us tantalizingly close to the goal of deriving patient-specific hematopoietic stem cells (HSCs). While directed differentiation and transcription factor-mediated conversion strategies have generated progenitor cells with multilineage potential, to date, therapy-grade...
Ex vivo tools for the clonal analysis of zebrafish hematopoiesis.
Nature Protocols 11(5):1007 (2016)
This protocol describes the ex vivo characterization of zebrafish hematopoietic progenitors. We show how to isolate zebrafish hematopoietic cells for cultivation and differentiation in colony assays in semi-solid media. We also describe procedures for the generation of recombinant zebrafish cyto...
Insight into GATA1 transcriptional activity through interrogation of cis elements disrupted in human erythroid disorders.
PNAS 113(16):4434 (2016)
Whole-exome sequencing has been incredibly successful in identifying causal genetic variants and has revealed a number of novel genes associated with blood and other diseases. One limitation of this approach is that it overlooks mutations in noncoding regulatory elements. Furthermore, the mechan...
Stress from Nucleotide Depletion Activates the Transcriptional Regulator HEXIM1 to Suppress Melanoma.
Molecular Cell 62(1):34 (2016)
Studying cancer metabolism gives insight into tumorigenic survival mechanisms and susceptibilities. In melanoma, we identify HEXIM1, a transcription elongation regulator, as a melanoma tumor suppressor that responds to nucleotide stress. HEXIM1 expression is low in melanoma. Its overexpression i...
Fish to Learn: Insights into Blood Development and Blood Disorders from Zebrafish Hematopoiesis.
Human Gene Therapy 27(4):287 (2016)
Since its introduction in early 1980s, the zebrafish (Danio rerio) has become an invaluable vertebrate animal model system to study many human disorders in almost all systems, from hepatic and brain pathology, to autoimmune and psychiatric disorders. Hematopoiesis between zebrafish and mammals i...
A Tribute to George Stamatoyannopoulos.
Human Gene Therapy 27(4):280 (2016)
GATA Factor-G-Protein-Coupled Receptor Circuit Suppresses Hematopoiesis.
Stem cell reports 6(3):368 (2016)
Hematopoietic stem cells (HSCs) originate from hemogenic endothelium within the aorta-gonad-mesonephros (AGM) region of the mammalian embryo. The relationship between genetic circuits controlling stem cell genesis and multi-potency is not understood. A Gata2 cis element (+9.5) enhances Gata2 exp...
A zebrafish melanoma model reveals emergence of neural crest identity during melanoma initiation.
Science 351(6272):aad2197 (2016)
The "cancerized field" concept posits that cancer-prone cells in a given tissue share an oncogenic mutation, but only discreet clones within the field initiate tumors. Most benign nevi carry oncogenic BRAF(V600E) mutations but rarely become melanoma. The zebrafish crestin gene is expressed embry...
Dynamic Control of Enhancer Repertoires Drives Lineage and Stage-Specific Transcription during Hematopoiesis.
Developmental Cell 36(1):9 (2016)
Enhancers are the primary determinants of cell identity, but the regulatory components controlling enhancer turnover during lineage commitment remain largely unknown. Here we compare the enhancer landscape, transcriptional factor occupancy, and transcriptomic changes in human fetal and adult hem...
Targeted Application of Human Genetic Variation Can Improve Red Blood Cell Production from Stem Cells.
Cell Stem Cell 18(1):73 (2016)
Multipotent and pluripotent stem cells are potential sources for cell and tissue replacement therapies. For example, stem cell-derived red blood cells (RBCs) are a potential alternative to donated blood, but yield and quality remain a challenge. Here, we show that application of insight from hum...
Generation of Parabiotic Zebrafish Embryos by Surgical Fusion of Developing Blastulae.
Journal of Visualized Experiments (112) (2016)
Surgical parabiosis of two animals of different genetic backgrounds creates a unique scenario to study cell-intrinsic versus cell-extrinsic roles for candidate genes of interest, migratory behaviors of cells, and secreted signals in distinct genetic settings. Because parabiotic animals share a c...
Identifying Novel Cancer Therapies Using Chemical Genetics and Zebrafish.
Advances in Experimental Medicine and Biology 916:103 (2016)
Chemical genetics is the use of small molecules to perturb biological pathways. This technique is a powerful tool for implicating genes and pathways in developmental programs and disease, and simultaneously provides a platform for the discovery of novel therapeutics. The zebrafish is an advantag...
Hematopoietic stem cells develop in the absence of endothelial cadherin 5 expression.
Blood 126(26):2811 (2015)
Rare endothelial cells in the aorta-gonad-mesonephros (AGM) transition into hematopoietic stem cells (HSCs) during embryonic development. Lineage tracing experiments indicate that HSCs emerge from cadherin 5 (Cdh5; vascular endothelial-cadherin)(+) endothelial precursors, and isolated population...
A Quantitative System for Studying Metastasis Using Transparent Zebrafish.
Cancer Research 75(20):4272 (2015)
Metastasis is the defining feature of advanced malignancy, yet remains challenging to study in laboratory environments. Here, we describe a high-throughput zebrafish system for comprehensive, in vivo assessment of metastatic biology. First, we generated several stable cell lines from melanomas o...
Generation of vascular endothelial and smooth muscle cells from human pluripotent stem cells.
Nature Cell Biology 17(8):994 (2015)
The use of human pluripotent stem cells for in vitro disease modelling and clinical applications requires protocols that convert these cells into relevant adult cell types. Here, we report the rapid and efficient differentiation of human pluripotent stem cells into vascular endothelial and smoot...
Mutation of kri1l causes definitive hematopoiesis failure via PERK-dependent excessive autophagy induction.
Cell Research 25(8):946 (2015)
Dysregulation of ribosome biogenesis causes human diseases, such as Diamond-Blackfan anemia, del (5q-) syndrome and bone marrow failure. However, the mechanisms of blood disorders in these diseases remain elusive. Through genetic mapping, molecular cloning and mechanism characterization of the z...
Epoxyeicosatrienoic acids enhance embryonic haematopoiesis and adult marrow engraftment.
Nature 523(7561):468 (2015)
Haematopoietic stem and progenitor cell (HSPC) transplant is a widely used treatment for life-threatening conditions such as leukaemia; however, the molecular mechanisms regulating HSPC engraftment of the recipient niche remain incompletely understood. Here we develop a competitive HSPC transpla...
TopBP1 Governs Hematopoietic Stem/Progenitor Cells Survival in Zebrafish Definitive Hematopoiesis.
PLoS Genetics 11(7):e1005346 (2015)
In vertebrate definitive hematopoiesis, nascent hematopoietic stem/progenitor cells (HSPCs) migrate to and reside in proliferative hematopoietic microenvironment for transitory expansion. In this process, well-established DNA damage response pathways are vital to resolve the replication stress, ...
Screening for melanoma resistance genes in vivo.
Pigment Cell & Melanoma Research 28(4):375 (2015)