Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1996-03-13
1998-11-03
Jones, W. Gary
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
4353201, 536 245, C07H 2104, C12N 1563
Patent
active
058310487
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a new gene, designated Grb3-3, its variants, and their uses, especially in anti-cancer gene therapy.
Various genes, called oncogenes and suppressor genes, are involved in the control of cell division. Among them, the ras genes and their products generally designated p21 proteins, play a key role in the control of cell proliferation in all eukaryotic organisms where they have been searched out. In particular, it has been shown that certain specific modifications of these proteins cause them to lose their normal control and lead them to become oncogenic. Thus, a large number of human tumours have been associated with the presence of modified ras genes. Likewise, an overexpression of these p21 proteins can lead to a deregulation of cell proliferation. An understanding of the exact role of these p21 proteins in cells, of their mode of operation and their characteristics therefore constitutes a major stake for the understanding and the therapeutic approach to carcinogenesis.
Various factors involved in the ras-dependent signalling pathway have been identified. Among these are the Grb2 gene which encodes a protein of 23-25 kDa having a SH3-SH2-SH3 structure (Lowenstein et al., Cell 70 (1992) 431; Matuoka et al., PNAS 89 (1992) 9015). The product of the Grb2 gene appears to interact with the tyrosine phosphorylated proteins via its SH2 domain, and with a factor for exchange of GDP of the SOS class via its SH3 domain (Egan et al., Nature 363 (1993) 45). It would thus be one of the components of the transformant activity of the product of the ras gene. The present invention derives from the demonstration of the cloning and characterization of an isoform of the Grb2 gene, designated Grb3-3, possessing a deletion in the SH2 domain. This gene is expressed in adult tissues: the corresponding mRNA is present in the form of a single band of 1.5 kb, and is translated into a 19 kDa protein. Because of its deletion in the SH2 domain, the product of the Grb3-3 gene is no longer capable of interacting with the tyrosine phosphorylated proteins (phosphorylated EGF receptor), but it retains the capacity to interact with the proline-rich domains of the SOS proteins. Because of its deletion, the product of the Grb3-3 gene is thus capable of preventing the cellular effects of the product of the Grb2 gene. The transfer of this gene in vivo, or of variants thereof, including antisense sequences, therefore makes it possible to interfere with the processes of proliferation, differentiation and/or cell death.
A first subject of the invention therefore relates to a nucleotide sequence comprising all or part of the Grb3-3 gene (sequence SEQ ID No. 1).
Another subject of the invention relates to a nucleotide sequence derived from the sequence SEQ ID No. 1 and capable of inhibiting, at least partially, the expression of the Grb2 or Grb3-3 protein. In particular, the invention relates to the antisense sequences whose expression in a target cell makes it possible to control the transcription of cellular mRNAs. Such sequences can for example be transcribed, in the target cell, into RNAs complementary to the cellular mRNAs Grb2 or Grb3-3 and thus block their translation into protein, according to the technique described in patent EP 140 308. Such sequences may consist of all or part of the nucleic sequence SEQ ID No. 1, transcribed in the reverse orientation.
As indicated above, Grb2 is a protein which is at least bifunctional, and which is anchored via its SH2 domain to specific sequences phosphorylated at the tyrosine, and via its two SH3 domains, to the exchange factors of the SOS family. Grb3-3 having lost its capacity to associate with proteins phosphorylated at the tyrosine can therefore only form a complex with the SOS proteins. Grb3-3 can therefore prevent the recruitment of the Grb2-SOS complex by the receptors of the self-phosphorylated growth factors or by associated proteins which are also phosphorylated at the tyrosine such as HSC or IRS1. Grb3-3 being capable of blocking this recruitment, it is cap
REFERENCES:
patent: 5434064 (1995-07-01), Schlessinger et al.
Suen et al. Mol. Cell. Biol. (1993) 13:5500-5512.
Watanabe et al. (1995) J. Biol. Chem. 270:13733-39.
Matuoka et al. (1992) Proc. Natl. Acad. Sci. USA 89:9015-19.
Lowenstein et al. (1992) Cell 70:431-42.
Schweighoffer Fabien
Tocque Bruno
Atzel Amy
Jones W. Gary
Rhone-Poulenc Rorer S.A.
LandOfFree
Grb3-3 cDNA and polypeptides does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Grb3-3 cDNA and polypeptides, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Grb3-3 cDNA and polypeptides will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-690626