+

WO1999014323A1 - Hereguline gamma - Google Patents

Hereguline gamma Download PDF

Info

Publication number
WO1999014323A1
WO1999014323A1 PCT/IB1997/001287 IB9701287W WO9914323A1 WO 1999014323 A1 WO1999014323 A1 WO 1999014323A1 IB 9701287 W IB9701287 W IB 9701287W WO 9914323 A1 WO9914323 A1 WO 9914323A1
Authority
WO
WIPO (PCT)
Prior art keywords
heregulin
sequence
hrg
nucleotide sequence
domain
Prior art date
Application number
PCT/IB1997/001287
Other languages
English (en)
Inventor
Urs Eppenberger
Fabrice Schoumacher
Heinz Müller
Original Assignee
Urs Eppenberger
Fabrice Schoumacher
Mueller Heinz
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Urs Eppenberger, Fabrice Schoumacher, Mueller Heinz filed Critical Urs Eppenberger
Priority to AU44699/97A priority Critical patent/AU4469997A/en
Publication of WO1999014323A1 publication Critical patent/WO1999014323A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/4756—Neuregulins, i.e. p185erbB2 ligands, glial growth factor, heregulin, ARIA, neu differentiation factor
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a novel protein of the heregulin family, particularly to a heregulin which is truncated in the epidermal growth factor (EGF)- like domain.
  • EGF epidermal growth factor
  • HRG Heregulins
  • Ig immuno- globulin
  • EGF epidermal growth factor
  • HRG glycosylated (30- 40 %) heparin-binding molecules (6,11,12) produced as precursors containing a transmembrane domain with a cyto- plasmatic tail of variable length. Their secretion occurs by insertion of the precursors into the plasma membrane and the release of the respective mature factor by spe ⁇ cific proteolysis. Lack of the transmembrane domain impairs secretion (9) .
  • ErbB-2 inactivation is a novel strategy to treat advanced breast cancer, since ErbB-2 overex- pression is correlated with poor prognosis (13) and loss of antiestrogen treatment efficacy in human breast cancer (HBC) (14) .
  • HRG have been implicated in the progression of the disease (15) .
  • One object of the present invention was to isolate novel heregulin variants as a basis for an improved treatment of e.g. human breast cancer.
  • a further object of the present invention was to determine the nucleic acid structure of such heregulin variants .
  • Another object of the present invention was to provide a diagnostic agent for cancer diagnosis.
  • Still another object of the present invention was to provide methods for the production of therapeutic agents and such therapeutic agents themselves, particularly for breast cancer therapy.
  • Yet another object of the present invention was to provide a method for the regulation of cell activity, of cell activation processes and/or the stimulation/inhibition of cell proliferation, particularly of cells lacking an ErbB receptor.
  • Said splicing variant is characterized by the coding sequence of amino acids 1 to 211 of heregulin terminated by a stop codon.
  • cod-ing sequence is flanked by a 568 bp 3' -untranslated region which has no correspondence in the cDNA sequences of heregulin- ⁇ as well as heregulin- ⁇ .
  • AAACTTTTCC CAAACCCGAT CCGAGCCCTT GGACCAAACT CGCCTGCGCC GAGAGCCGTC 420
  • AAACTAGTCC TTCGGTGTGA AACCAGTTCT GAATACTCCT CTCTCAGATT CAAGTGGTTC 660
  • the novel heregulin variant which is truncated in the EGF-like domain, provides a useful means to generate antibodies which do selectively recognize the specific end of the respective heregulin variant in the EGF-like domain.
  • the heregulin needed to generate such antibodies according to known methods can be produced by recombinant methods using either the cDNA or a respective part of the genomic heregulin DNA.
  • Antibodies can also be generated against synthetic peptides having the same sequence as HRG- ⁇ protein sequence.
  • the heregulin- ⁇ specific probes for e.g. cancer diagnostics and antisense therapy agents can be generated according to general methods known in the art such as synthetic methods.
  • the heregulins of the present invention provide a useful means to produce specific monoclonal antibodies. Such antibodies allow the detection of persons with an enhanced risk for breast cancer. Another method to determine such persons is the determination of the heregulin- ⁇ concentration by means of probes constructed on the basis of the 3' untranslated region adjacent to the exon 6 sequence, i.e. probes derived from the sequence information of nucleo- tides 637 to 1204 of Table I, particularly nucleotides 637 to 1186. Since nucleotides 635 and 636 are also different from the corresponding nucleotides in HRG alpha and beta, probes including these nucleotides could be used as well.
  • antisense therapy agents can be prepared for e.g. breast cancer therapy, said agents being RNA or DNA antisense oligonucleotides selected from the heregulin- ⁇ specific region, i.e. the 3' -untranslated region, particularly from the exon 6 adjacent part of said region.
  • Antisense molecules can also be obtained from expression vectors (plasmids) comprising a sequence of interest introduced into the target cells where they will allow the transcription of large (up to several kilobase) RNA molecules of antisense polarity.
  • the heregulin- ⁇ is an extracellular cell stimulator, it is a suitable tool for the detection of growth inhibitors or for the stimulation of the cell proliferation, particularly for cells lacking an ErbB receptor. Such stimulation can be performed either by recombinant techniques or by administration of the protein.
  • HRG- ⁇ thus is also suitable as therapeutic agents for diseases connected with proliferation prob- lems.
  • Cells with proliferation problems are e.g. involved in osteoporosis.
  • nucleotide sequence of the present invention still allowing the base pair formation under suitable, usually stringent, conditions are considered as falling within the scope of the present invention.
  • sequences generally are about 90 % homo- logue to the sequence outset in Table I.
  • Figures 1 to 3 show a partial sequence alignment of HRG- ⁇ with HRG- and HRG- ⁇ and northern blot analysis of HRG- ⁇ expression.
  • Figure 1 represents HRG- ⁇ cDNA sequence (nucleotides 613 to 687 relative to the initiation codon) together with the deduced amino acid sequence (single letter code) are aligned with HRG- ⁇ and HRG- ⁇ isoform nu- cleic acid and peptide sequences.
  • the stop codon in HRG- ⁇ is marked with *. Cysteines in the EGF-like motif are shown in boldface.
  • Figure 2 shows the northern blot analysis of HRG- ⁇ expression in various HBC cell lines.
  • the northern blotting was performed using a HRG- ⁇ specific radiola- beled probe. 20 ⁇ g total RNA were analysed.
  • 1-2 MDA-MB- 231 and MCF-7 cells (Mason Research Institute, Rockville, MD) .
  • 3-6 HS578-T, SKBRIII, ZR-75-1 and T47-D cells, re- spectively (American Type Culture Collection, Rockville, MD) .
  • Filled arrow HRG- ⁇ mRNA, open arrow : position of the 2.3kb rRNA.
  • Figure 3 shows detection of HRG- ⁇ in biopsies.
  • Total RNA was extracted from tumor tissues and subjected to RT-PCR.
  • Upper panel PCR amplification of GAPDH
  • FIG. 4 showing the ErbB-2 tyrosine phospho- rylation status.
  • SKBR-3 cells (5 x 10 7 cells) were either mock stimulated (control) or stimulated with HRG- ⁇ l (1 nM) or HRG- ⁇ (30 pM) for 8 minutes at 37°C.
  • Cells were lysed (41) and ErbB-2 was immunoprecipitated with 10 ⁇ g polyclonal ErbB-2 antibody (Santa Cruz Biotechnology, CA) .
  • Immunoprecipitates were analysed by western blotting. The ErbB-2 protein was stained by incubation of the blot with the antibody used for immunoprecipitation, whereas the ErbB-2 phosphotyrosine content was revealed with PY20 antibody (Santa Cruz Biotechnology, CA) .
  • Figure 5 represents growth promotion by recombinant HRG- ⁇ .
  • Figure 6 represents MAP kinase stimulation. MCF-7 cells (10' cells) were incubated in serum-free medium for 24 hours and stimulated for the time indicated either with EGF (lnM) or HRG- ⁇ (30pM) .
  • MAP kinase activ- ity was measured as described (43) (22) using a synthetic peptide (Amersham, UK) based on the EGF receptor Threonine669 phosphorylation site as substrate.
  • FIGS. 7 and 8 show subcellular localization of HRG- ⁇ . MDA-MB-231 cells were electroporated with pEGFP-Cl derivatives, seeded onto sterile glass slides and analysed after 24 hours incubation without fixation. Figure 7 depicts the HRG exons together with the restriction sites used for the construction of the deletion mutants. Hatched boxes: GFP, black box: SV40 nuclear localization sequence. Gray boxes: HRG sequences. Circles: cysteines.
  • Figure 8 shows the fluorescence of MDA-MB-231 cells transfected with: a: pEGFP, b: pEGFP/NLS, c: pEGFP/HRG- ⁇ SaCII, pEGFP/HRG- ⁇ Spel or pEGFP/HRG- ⁇ BclI, d: pEGFP/HRG- ⁇ BbsI, pEGFP/HRG- ⁇ Xmnl, pEGFP/HRG- ⁇ XhoI, pEGFP/HRG- ⁇ or pEGFP/HRG- ⁇ .
  • Figure 9 shows HRG- ⁇ stimulation by estrogen and tamoxifen.
  • MCF-7 cells kept on serum free medium were stimulated with O.lnM estrogen (Fig. 9 A) or 0.1 ⁇ M Tamoxifen (Figure 9 B) for the time indicated.
  • Total RNA (20 ⁇ g) was analysed by northern blotting with a HRG- ⁇ specific probe. Expression was determined by densitometry and is expressed as arbitrary units.
  • a cDNA library obtained from the HBC cell line MDA-MB-231 was screened and a l. ⁇ kb clone of a novel heregulin splicing variant (see Table I) which encodes an open reading frame of 211 amino acids containing the Ig-like, glycosylation domain and part of the EGF-like domain identical with the corresponding HRG- ⁇ and ⁇ domains (6, 17) was isolated.
  • the library was constructed with 10 ⁇ g polyA RNA obtained from MDA-MB-231 cells.
  • the cDNA cloning into the ⁇ ZAPII vector (Stratagene, CA) was performed according to the manufacturer instructions.
  • a stop codon interrupts the EGF-like motif after the fourth cysteine (Fig. 1) .
  • the coding sequence is flanked by a 568bp 3' -untranslated region ending with a poly (A) tail.
  • Said heregulin variant is denominated herein furtheron HRG- ⁇ since it is the third known sequence variation for the EGF-like motif and it is the first HRG splicing variant containing a trun- cated EGF-like domain.
  • RNA samples Frozen biopsies, pulverized in liquid nitrogen or cell cultures were subjected to total RNA extraction using the RNeasy kit (Qiagen, FRG) .
  • cDNA was synthesized from 1 ⁇ g total RNA at 42 °C for 60 minutes with MuMLV reverse transcriptase (Promega, I) in a 20 ⁇ l volume.
  • the cDNA was diluted 5 fold and 1 ⁇ l was used as template for a 25 cycle control PCR reaction with primers pel (5' GGTGAAGGTCGGAGTCAACGG) and pc2
  • GAPDH glyceraldehyde 3 phosphate deshydrogenase
  • the amount of GAPDH product was calculated by subjecting 5 ⁇ l control PCR to gel electrophoresis . Digital imaging of the gel stained with a 1/10,000 dilution of SYBRTMGreen I DNA stain (Molecular Probes, OR) was performed with the GelDoc system (BioRad, CA) . The cDNA amount of each sample was calculated according to the relative amount of the GAPDH product. HRG- ⁇ cDNA amplifi- cation was performed with primers p3
  • PCR was carried out in buffer H (Invitrogen, CA) with 1 unit Taq DNA polymerase (Boehringer, FRG) in 50 ⁇ l reaction volume. PCR reactions were cycled 94 °C 2 min, 94°C 30 sec, 55°C 30 sec, 72°C 1 min for 25 cycles with a final extension step at 72°C for 5 min. These conditions did not allow any fluorescence-based detection of amplification products (data not shown) . Negative controls were made with RNA not subjected to reverse transcription and with no template.
  • PCR products were identified by southern blotting with probes obtained by PCR with the corresponding primer pair using plasmid pBluescript/HRG- ⁇ as template. Labeling of probes was performed by nick translation with [ ⁇ - 32 P]-dCTP (Amersham, UK) using the Ready-to-go labeling kit (Pharmacia, Sweden) . Unincorporated nucleotides were removed by column purification (Push column, Stratagene) . The PCR reactions were separated by electrophoresis on 2% agarose gels, at 5 V/cm and transferred overnight by alkaline blotting to HybondN+ membranes (Amersham, UK) which were hybridized at 68 °C overnight (44) .
  • the membranes were subsequently washed at 50°C for 15 min at low stringency and 42°C for 15 min at high stringency. Blots were exposed to phosphor storage screens for 1-2 days. Data were acquired with a Phosphorlmager device and quantified with the ImageQuant analysis program (Molecular Dynamics, CA) .
  • the DNA sequence linked to exon 6 (6) in HRG- ⁇ was characterized by PCR using a primer pair encompassing the stop codon to amplify genomic DNA from MDA-MB-231 and MCF-7 cells. The results revealed that the HRG- ⁇ specific sequence corresponds to the intron located 3' to exon 6 (herein furtheron referred to as intron 6) .
  • HRG- ⁇ levels were measured for HRG- ⁇ expression levels by semi-quantitative RT-PCR (see above) and compared to those of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as internal control (Fig. 3) .
  • GPDH glyceraldehyde-3-phosphate dehydrogenase
  • Fig. 3 HRG- ⁇ levels of HBC biopsies ranged from very high to low and did not correlate with the expression of HRG ⁇ / ⁇ (data not shown) , suggesting an independent transcriptional regulation of this splicing variant.
  • MCF-7 cells were transiently transfected with an expression vector carrying the HRG- ⁇ sequence. Re- striction sites were introduced in the HRG- ⁇ sequence by PCR amplification using primers p3 (5' TCCGTCTCCGCGAGACGGGA TCCGAGCGCAAAGAAGGC) and p4 (5' CATAAGCGACACACAGGAT CCGTCGACGAATTCTTACAAGCACAAGTATCTC) and plasmid pBluescript/HRG- ⁇ as template.
  • the primer p3 introduces a BamHl site (boldface) at the site of the ATG codon whereas primer p4 introduces EcoRI, Sail and BamHl sites (boldface) 3' to the stop codon of HRG- ⁇ (underlined) .
  • the PCR product was cloned into plasmid pCRII (Invitrogen, CA) , producing pCRII/HRG- ⁇ .
  • a BamHI-Sall restriction fragment was ligated into the bacterial expression vector pQE30 (Qiagen, FRG) resulting in pQE30/HRG- ⁇ .
  • E.coli XL1 Blue cells transformed with plasmid pQE30/HRG- ⁇ , were grown overnight with shaking at 37 °C in LB medium supplemented with lOO ⁇ g/ml ampicillin.
  • the culture was diluted 100 fold with fresh medium and grown to an ODgoo °f !• Recombinant protein expression was induced by addition of IPTG (0.4mM final concentration).
  • the cells were grown for an additional 5 hours, centrifuged and the cell pellets were stored frozen at -70°C.
  • Inclusion bodies were solubilized in 6M urea and purified over a cation exchange column (SP Sepharose Fast Flow, Pharma- cia, Sweden) .
  • An enhanced Green Fluorescent Protein (EGFP) fusion expression vector was obtained by inserting the BamHI-Sall restriction fragment from pQE30/HRG- ⁇ into the Bglll and Sail sites of plasmid pEGFP-Cl (Clontech, CA) , in frame with the EGFP coding sequence, resulting in pEGFP/HRG- ⁇ .
  • HBC cell lines were transfected by electro- poration using the Gene Pulser II (BioRad) .
  • 10 7 cells were trypsinyzed, resuspended in 500 ⁇ l complete growth medium with 20-40 ⁇ g plasmid DNA and subjected to a sin- gle pulse (1350) V/cm, 960 ⁇ F, no parallel resistance) .
  • the cells were kept on ice for 10 min, resuspended in complete growth medium and seeded on sterile glass slides. After 24 hour, unfixed cells were observed with a Zeiss Axioskop microscope. GFP fluorescence was observed with FITC filters. Image acquisition was performed with a CCD camera using the MacProbe program (Perceptive Scientific Instruments Inc., UK).
  • GFP-fusion deletion mutants were obtained as follows: pEGFP/HRG- ⁇ BbsI and pEGFP/HRG- ⁇ Spel were constructed from plasmid pEGFP/HRG- ⁇ by line ⁇ arisation at the appropriate restriction sites, fill-in with T4 DNA polymerase and subsequent digestion with Smal and religation. Deletion mutant pEGFP/HRG- ⁇ Xmnl was obtained by double digestion with XmnI and Smal and religation.
  • Deletion mutants pEGFP/HRG- ⁇ XhoI, pEGFP/HRG- ⁇ BclI were obtained by double digestion with Xhol/Sall and BamHI/BclI respectively.
  • the pEGFP/HRG ⁇ SacII construct was obtained by SacII digestion and subsequent religa- tion.
  • pEGFP/HRG- ⁇ was obtained by prior isolation of a HRG- ⁇ PCR product obtained from MDA-MB-231 cell cDNA : With the primer pair p7 (3 'AAACTAGTCCTTCGGTG) and p8 ( 3 ' GGAATTCACATGA TGCCGACCACAAGGA) a 632 bp fragment was amplified, cloned into pCRII.
  • Plasmid pEGFP/NLS was obtained by ligating a phosphorylated synthetic oligonucleotide linker (pNl 3 ' TCGATATCCAAAGAAGA AGCGCAAGGTGCA and pN2 3' CCTTGCGCTTCTTCTTTGGATA) into plasmid pEGFP-Cl digested with Xhol and Pstl.
  • the Bcll-Bbsl domain may exert a specific biological function within the nucleus (Fig. 8, d) .
  • the subnuclear localization obtained with HRG sequences fused to GFP (Fig. 8, d) is reminiscent of the cell cycle dependent, nuclear dot pattern observed with human Rad51 (32) and BRCA1 distribution (33) and are consistent with the findings that HRG alter the cell cycle (15, 34, 35) .
  • heregulin- ⁇ is a suitable agent for the treatment of diseases connected with reduced cell proliferation such as e.g. osteoporosis. Such treatment can be performed by direct admini- stration of the heregulin or by transfection of heregulin deficient cells.
  • Heregulin- ⁇ is also a very useful means to produce selective antibodies due to the truncated EGF-do- main resulting in a heregulin- ⁇ -specific recognition site.
  • Such antibodies themselves are very useful diagnostic agents, allowing the determination of e.g. persons with an enhanced risk for breast cancer.
  • Another agent for the determination of e.g. persons with an enhanced risk for breast cancer is an oligonucleotide comprising a sequence, which is at least partially derived from the 3' -untranslated region adjacent to exon 6 of the heregulin ⁇ -sequence of Table I.
  • Said sequence of the oligonucleotide, which is derived from the 3' ntranslated region addressed above hybridizes under stringent conditions with the heregulin- ⁇ RNA.
  • Such sequences generally are about at least 90 % homologue to the respective sequence of Table I.
  • Heregulin- ⁇ can also be used to evaluate and search for growth inhibitors suitable as effective sub- stances in cancer therapy, e.g. alone or in combination with other effective substances.
  • Site directed mutagenesis of the coding sequence allows to produce recombinant proteins that use the same pathway (bind to the same receptor (s) ) but do not trigger downstream signalling, thereby competing with HRG- ⁇ .
  • the cDNA sequence of Table I provides an important means for the production of selective antisense therapy agents which - for good selectivity - are made on the basis of the 3' -untranslated region, preferably of the intron 6 like sequence adjacent to exon 6.
  • Heregulin- ⁇ can also be used to generally stimulate cells lacking an ErbB receptor either as therapeutical agent or in cell cultures. Besides of the administration of the protein itself, cells can be transfected with the heregulin- ⁇ DNA.
  • the Ig-like domain is a very important sequence for the nuclear or subnuclear localization.
  • This sequence thus can be used as such or modified to regulate cell active processes.
  • Such processes can e.g. be the heregulin- ⁇ production, which can be regulated by modifications performed in the Ig-like domain such as deletions, substitutions etc., or the Ig-like domain or its nucleotide sequence, respectively, can as such or in modified form be connected with suitable cell active proteins or nucleotide sequences thereof, respectively.
  • the oligonucleotides of the present invention usually have an interesting part of a length of 10 or more bases. In this range a homology about 90 % usually is sufficient to get hybridization under suitably selective conditions.
  • Interesting part means a contiguous part of the oligonucleotide that is intended to hybridize with the sequence of interest. It is general knowledge that hybridization is obtained if at least one contiguous part(s) of the oligonucleotide has at least about 90 % homology with a sequence of interest and that the selectivity is enhanced with longer "interesting parts”.
  • Oligonucleotides can be made by chemical or enzymatic synthesis. Large nucleotide sequences advanta ⁇ geously can be made by current Polymerase Chain Reaction techniques. An Ig-like domain or modified Ig-like domain comprising protein can also be used for the nuclear import of cell active molecules.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne des variantes de l'héréguline (HRG-gamma) dont le domaine du type EGF est tronqué, et des séquences d'ADN codant lesdites variantes. L'invention porte également sur l'application pharmaceutique desdites hérégulines et sur des agents thérapeutiques et diagnostiques dérivés desdites hérégulines ou de leurs séquences nucléotidiques.
PCT/IB1997/001287 1997-09-17 1997-10-17 Hereguline gamma WO1999014323A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU44699/97A AU4469997A (en) 1997-09-17 1997-10-17 Heregulin-gamma

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GHPCT/IB97/01115 1997-09-17
IB9701115 1997-09-17

Publications (1)

Publication Number Publication Date
WO1999014323A1 true WO1999014323A1 (fr) 1999-03-25

Family

ID=11004605

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB1997/001287 WO1999014323A1 (fr) 1997-09-17 1997-10-17 Hereguline gamma

Country Status (2)

Country Link
AU (1) AU4469997A (fr)
WO (1) WO1999014323A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001001748A3 (fr) * 1999-07-02 2001-07-26 Genentech Inc Composés se liant à her2
EP1456239A4 (fr) * 2001-07-31 2005-04-27 Univ Wayne State Proteines hybrides a domaine se liant a l'heparine de neureguline pour cibler des proteoglycanes de sulfate d'heparane
US9828635B2 (en) 2011-10-06 2017-11-28 Aveo Pharmaceuticals, Inc. Predicting tumor response to anti-ERBB3 antibodies

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994028133A1 (fr) * 1993-05-21 1994-12-08 Amgen Inc. FACTEURS DE DIFFERENCIATION RECOMBINES DE $i(NEU)
WO1995002052A1 (fr) * 1993-07-06 1995-01-19 F. Hoffmann-La Roche Ag Mammamoduline proteine specifique des cellules tumorales mammaires hormono-independantes

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994028133A1 (fr) * 1993-05-21 1994-12-08 Amgen Inc. FACTEURS DE DIFFERENCIATION RECOMBINES DE $i(NEU)
WO1995002052A1 (fr) * 1993-07-06 1995-01-19 F. Hoffmann-La Roche Ag Mammamoduline proteine specifique des cellules tumorales mammaires hormono-independantes

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DUANZHI WEN ET AL.: "Neu differentition factor: A transmembrane glycoprotein containing an EGF domain and an Immunoglobulin homology unit", CELL, vol. 69, 1 May 1992 (1992-05-01), NA US, pages 559 - 572, XP002002185 *
DUANZHI WEN ET AL.: "Structural and functional aspects of the multiplicity of Neu differentiation factors", MOLECULAR AND CELLULAR BIOLOGY, vol. 14, no. 3, March 1994 (1994-03-01), WASHINGTON US, pages 1909 - 1919, XP002064746 *
SCHAEFER, GABRIELE ET AL: ". gamma.- Heregulin: a novel heregulin isoform that is an autocrine growth factor for the human breast cancer cell line, MDA-MB-175", ONCOGENE (1997), 15(12), 1385-1394 CODEN: ONCNES;ISSN: 0950-9232, 1997, XP002064745 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001001748A3 (fr) * 1999-07-02 2001-07-26 Genentech Inc Composés se liant à her2
US6987088B2 (en) 1999-07-02 2006-01-17 Genentech, Inc. Compounds that bind HER2
EP1456239A4 (fr) * 2001-07-31 2005-04-27 Univ Wayne State Proteines hybrides a domaine se liant a l'heparine de neureguline pour cibler des proteoglycanes de sulfate d'heparane
AU2002322762B2 (en) * 2001-07-31 2008-10-16 Wayne State University Hybrid proteins with neuregulin heparin-binding domain for targeting to heparan sulfate proteoglycans
US7527794B2 (en) 2001-07-31 2009-05-05 Wayne State University Hybrid proteins with neuregulin heparin-binding domain for targeting to heparan sulfate proteoglycans
US9828635B2 (en) 2011-10-06 2017-11-28 Aveo Pharmaceuticals, Inc. Predicting tumor response to anti-ERBB3 antibodies

Also Published As

Publication number Publication date
AU4469997A (en) 1999-04-05

Similar Documents

Publication Publication Date Title
Higashiyama et al. A novel brain-derived member of the epidermal growth factor family that interacts with ErbB3 and ErbB4
Chang et al. Ligands for ErbB-family receptors encoded by a neuregulin-like gene
Alves et al. Identification of two novel, kinase‐deficient variants of discoidin domain receptor 1: differential expression in human colon cancer cell lines
Schaefer et al. γ-Heregulin: a novel heregulin isoform that is an autocrine growth factor for the human breast cancer cell line, MDA-MB-175
Noma et al. Molecular cloning and structure of the human transforming growth factor-β2 gene promoter
US20030232755A1 (en) Cripto mutant and uses thereof
HUT77578A (hu) A 10-es számú fibroblaszt növekedési faktor (FGF-10)
US5538861A (en) DNA encoding a stimulating factor for the axl receptor
DE69530944T2 (de) Keratinozyten-wachstumsfaktor 2
JPH10502534A (ja) 結合組織増殖因子−２
US5981201A (en) Methods of detection and treatment of breast cancer
US20020160451A1 (en) Novel orphan receptors
US7705117B2 (en) EGFH2 genes and gene products
Shinozaki et al. Changes in pre-mRNA splicing factors during neural differentiation in P19 embryonal carcinoma cells
WO1999014323A1 (fr) Hereguline gamma
Shu et al. Dissecting the activating mutations in v-erbB of avian erythroblastosis virus strain R
Hu et al. Human fibroblast growth factor-18 stimulates fibroblast cell proliferation and is mapped to chromosome 14p11
US6075008A (en) Method for promoting cell proliferation and growth
KR101037384B1 (ko) 골다공증의 예방 또는 치료 활성을 갖는 물질의 스크리닝 방법
Delhanty et al. Cloning and characterization of the rat gene for the acid-labile subunit of the insulin-like growth factor binding protein complex
Jones et al. Identification and characterisation of chicken cDNAs encoding the endothelial cell-specific receptor tyrosine kinase Tie2 and its ligands, the angiopoietins
JPH11506919A (ja) 繊維芽細胞増殖因子１５
EP0504914A2 (fr) Procédé pour la détection des substances neuroactives
WO2001014415A2 (fr) Gènes egfh2 et produits géniques
Hoff III et al. DBI-1, a novel gene related to the notch family, modulates mitogenic response to insulin-like growth factor 1

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BR CA CN IL MX US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载