Growth factors induce a variety of cellular responses including proliferation, survival and migration but their final target genes remain for the most part unknown. In this proposal, we will focus on identifying targets for platelet derived growth factors (PDGFs) which are involved in various aspects of muscle and cardiovascular development, as well as upstream factors that control their expression. We will use a gene trap approach in ES cells to identify and clone genes whose expression is induced or repressed by PDGF, to study their expression pattern, and to assess the consequences of loss of function in mutant mice. We will concentrate our efforts on genes that are expressed or result in mutant phenotypes in cell types which are deficient in PDGF or PDGF receptor mutant mice. We will study PDGF-dependent gene transcription in cells isolated from somites and presomitic mesoderm, in which PDGF has been implicated for proper patterning, using subtraction-suppression PCR and hybridization to """"""""DNA chip"""""""" microarrays. To identify factors that are genetically upstream of PDGF, we will use new gene trap vectors that activate a gene constitutively. The PDGF and PDGF receptor genes will be targeted with a promoterless reporter gene, allowing identification of PDGF upstream factors by increased expression of the reporter. Expression of the trapped genes will be disrupted following Cre recombination, resulting in loss of function mutations. This methodology will allow us to identify PDGF upstream factors as well as their normal physiological role. These studies should help us understand growth factor regulatory mechanisms, and provide information on the specificity and interplay of growth factor signaling pathways in physiological processes.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD024875-11
Application #
2761350
Study Section
Human Embryology and Development Subcommittee 1 (HED)
Program Officer
Moody, Sally Ann
Project Start
1989-01-01
Project End
2003-12-31
Budget Start
1999-01-01
Budget End
1999-12-31
Support Year
11
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109
Zou, Hongyan; Feng, Rui; Huang, Yong et al. (2015) Double minute amplification of mutant PDGF receptor ? in a mouse glioma model. Sci Rep 5:8468
Friedel, Roland H; Friedel, Caroline C; Bonfert, Thomas et al. (2013) Clonal expansion analysis of transposon insertions by high-throughput sequencing identifies candidate cancer genes in a PiggyBac mutagenesis screen. PLoS One 8:e72338
Olson, Lorin E; Soriano, Philippe (2011) PDGFR? signaling regulates mural cell plasticity and inhibits fat development. Dev Cell 20:815-26
Friedel, Roland H; Soriano, Philippe (2010) Gene trap mutagenesis in the mouse. Methods Enzymol 477:243-69
Raymond, Christopher S; Soriano, Philippe (2010) ROSA26Flpo deleter mice promote efficient inversion of conditional gene traps in vivo. Genesis 48:603-6
Wassarman, Paul M; Soriano, Philippe M (2010) Guide to techniques in mouse development. Preface. Methods Enzymol 476:xix
Olson, Lorin E; Soriano, Philippe (2009) Increased PDGFRalpha activation disrupts connective tissue development and drives systemic fibrosis. Dev Cell 16:303-13
Bush, Jeffrey O; Soriano, Philippe (2009) Ephrin-B1 regulates axon guidance by reverse signaling through a PDZ-dependent mechanism. Genes Dev 23:1586-99
Schmahl, Jennifer; Rizzolo, Kamran; Soriano, Philippe (2008) The PDGF signaling pathway controls multiple steroid-producing lineages. Genes Dev 22:3255-67
Davy, Alice; Soriano, Philippe (2007) Ephrin-B2 forward signaling regulates somite patterning and neural crest cell development. Dev Biol 304:182-93

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