Congenital myeloid failure syndromes are a heterogeneous group of rare hereditary disorders associated with considerable morbidity. Despite recent insights into the genetic basis for a subset of these disorders, the molecular pathogenesis of others remains unknown. Our goal is to use a forward genetic approach in a model vertebrate, the zebrafish, to develop animal models of hereditary myeloid failure. Comprehensive characterization of these models, coupled with their genetic elucidation, will lead to new knowledge about the molecular pathogenesis of these disorders, aiding their diagnosis, treatment and possibly prevention. A group of zebrafish mutants with myeloid failure has been generated by ethylnitrosourea mutagenesis and genetic screening. Ten stable mutant pedigrees have been recovered to date (a recovery rate of 83%), and >15 more are at various stages of recovery. This project aims to (1) characterize the mutants descriptively using microscopy, histology, whole mount in situ hybridization gene expression analysis, and transplantation assays; (2) position all the mutants on the zebrafish genome map in their respective linkage groups; (3) positionally clone 6 of the mutants, prioritized for the particular biological interest of their phenotypes. Having identified the genetic basis of these particularly interesting myeloid-failure mutants, we will better understand the function of these genes in the molecular and cellular mechanisms of myeloid cell development. Because of the unbiased nature of generating the mutants, some of them are expected to reveal completely new insights into the causes of congenital myeloid failure. Other mutants, found to have novel mutations in genes already associated with myelopoiesis, will likely reveal new insights into the molecular pathogenesis of myeloid cell disease and normal myeloid cell proliferation, differentiation, survival and function. A better understanding of normal myeloid cell development will also help understand the basis for the dysregulation of these processes in myeloproliferative and leukemic diseases.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL079545-06
Application #
7473910
Study Section
Special Emphasis Panel (ZHL1-CSR-D (S1))
Program Officer
Qasba, Pankaj
Project Start
2004-09-30
Project End
2010-07-31
Budget Start
2008-08-01
Budget End
2010-07-31
Support Year
6
Fiscal Year
2008
Total Cost
$256,009
Indirect Cost
Name
Walter and Eliza Hall Institute Medical Research
Department
Type
DUNS #
753236256
City
Parkville
State
Country
Australia
Zip Code
VIC, -3052
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Keightley, Maria-Cristina; Crowhurst, Meredith O; Layton, Judith E et al. (2013) In vivo mutation of pre-mRNA processing factor 8 (Prpf8) affects transcript splicing, cell survival and myeloid differentiation. FEBS Lett 587:2150-7
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Pase, Luke; Layton, Judith E; Wittmann, Christine et al. (2012) Neutrophil-delivered myeloperoxidase dampens the hydrogen peroxide burst after tissue wounding in zebrafish. Curr Biol 22:1818-24
Pase, Luke; Lieschke, Graham J (2011) Discerning different in vivo roles of microRNAs by experimental approaches in zebrafish. Methods Cell Biol 104:353-78
Keightley, Maria-Cristina; Layton, Judith E; Hayman, John W et al. (2011) Mediator subunit 12 is required for neutrophil development in zebrafish. PLoS One 6:e23845
Ellett, Felix; Pase, Luke; Hayman, John W et al. (2011) mpeg1 promoter transgenes direct macrophage-lineage expression in zebrafish. Blood 117:e49-56
Ellett, Felix; Kile, Benjamin T; Lieschke, Graham J (2009) The role of the ETS factor erg in zebrafish vasculogenesis. Mech Dev 126:220-9
Pase, Luke; Lieschke, Graham J (2009) Validating microRNA Target Transcripts Using Zebrafish Assays. Methods Mol Biol 546:227-40

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