Intracellular Mediators of the Growth Response to Pdgf

Stiles, Charles D.

Abstract

Platelet-derived growth factor (PDGF) is a connective tissue mitogen which may play a role in the physiology of wound healing in vivo. Addition of PDGF to growth arrested 3T3 cells results in a prompt (within 40 ft.) induction of one or several specific poly (A)+ mRNAs. Induction of these mRNAs is not a mere consequence of cell growth. Rather a variety of data suggest that these mRNAs (or their translation products) function as stable, intracellular mediators of the growth response to PDGF. My laboratory has cloned and isolated cDNA probes directed to PDGF-inducible mRNA sequences within Balb/c-3T3 cells. This application focuses on the regulation of these mRNAs and function of their translation products in the mitogenic response. Manual microinjection procedures and recombinant DNA technology will be applied towards 3 broad objectives. The first objective is molecular characterization of PDGF-inducible gene sequences. """"""""Hybridization selection"""""""" and """"""""translation arrest"""""""" techniques will identify the proteins which correspond to each PDGF-inducible cDNA probe. The structural genes corresponding to our cDNA probes will be isolated from a cloned genomic library of Balb/c-3T3 cell DNA. The second objective is to establish the function of PDGF-inducible gene products in the mitogenic response of 3T3 cells. This objective will be approached i. by transfection of the cloned PDGF-inducible structural genes into normal 3T3 cells and ii. by manual microinjection of hybridization-selected mRNA into quiescent 3T3 cells. The third objective is to determine whether these gene sequences function as a common intracellular mediator of the growth response in other proliferating systems. Molecular hybridization techniques will be used to screen for expression of PDGF-inducible gene sequences within epithelioid cells and within connective tissue tumors. Additionally, tumor promotors and other mitogenic agents will be screened for their ability to modulate expression of these gene sequences.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of General Medical Sciences (NIGMS)
• Type: Research Project (R01)
• Project #: 5R01GM031489-03
• Application #: 3279518
• Study Section: Molecular Biology Study Section (MBY)
• Project Start: 1983-02-01
• Project End: 1986-01-31
• Budget Start: 1985-02-01
• Budget End: 1986-01-31
• Support Year: 3
• Fiscal Year: 1985

Institution

• Name: Dana-Farber Cancer Institute
• DUNS #: 149617367
• City: Boston
• State: MA
• Country: United States

Publications

Guha, A; Dashner, K; Black, P M et al. (1995) Expression of PDGF and PDGF receptors in human astrocytoma operation specimens supports the existence of an autocrine loop. Int J Cancer 60:168-73

Segal, R A; Pomeroy, S L; Stiles, C D (1995) Axonal growth and fasciculation linked to differential expression of BDNF and NT3 receptors in developing cerebellar granule cells. J Neurosci 15:4970-81

Wang, C; Stiles, C D (1994) Platelet-derived growth factor alpha receptor gene expression: isolation and characterization of the promoter and upstream regulatory elements. Proc Natl Acad Sci U S A 91:7061-5

Segal, R A; Goumnerova, L C; Kwon, Y K et al. (1994) Expression of the neurotrophin receptor TrkC is linked to a favorable outcome in medulloblastoma. Proc Natl Acad Sci U S A 91:12867-71

Shamah, S M; Stiles, C D; Guha, A (1993) Dominant-negative mutants of platelet-derived growth factor revert the transformed phenotype of human astrocytoma cells. Mol Cell Biol 13:7203-12

Wang, C; Shamah, S M; Stiles, C D (1992) Recombinant PDGF from lower vertebrates: receptor binding and immunochemical analysis with metabolically labeled growth factor. Growth Factors 7:279-88

Oquendo, P; Alberta, J; Wen, D Z et al. (1989) The platelet-derived growth factor-inducible KC gene encodes a secretory protein related to platelet alpha-granule proteins. J Biol Chem 264:4133-7

Hall, D J; Stiles, C D (1987) Platelet-derived growth factor-inducible genes respond differentially to at least two distinct intracellular second messengers. J Biol Chem 262:15302-8

Rollins, B J; Morrison, E D; Stiles, C D (1987) A cell-cycle constraint on the regulation of gene expression by platelet-derived growth factor. Science 238:1269-71

Zumstein, P; Stiles, C D (1987) Molecular cloning of gene sequences that are regulated by insulin-like growth factor I. J Biol Chem 262:11252-60