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WO2001040300A2 - Suppresseur de tumeurs p14 (arf) resistant a la protease - Google Patents

Suppresseur de tumeurs p14 (arf) resistant a la protease Download PDF

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Publication number
WO2001040300A2
WO2001040300A2 PCT/DE2000/004257 DE0004257W WO0140300A2 WO 2001040300 A2 WO2001040300 A2 WO 2001040300A2 DE 0004257 W DE0004257 W DE 0004257W WO 0140300 A2 WO0140300 A2 WO 0140300A2
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Prior art keywords
arf
nucleic acid
protein
acid molecule
peptide
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German (de)
English (en)
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WO2001040300A8 (fr
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Pidder JANSEN-DÜRR
Dominique Duro
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Deutsches Krebsforschungszentrum DKFZ
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Deutsches Krebsforschungszentrum DKFZ
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    • 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
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4703—Inhibitors; Suppressors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide

Definitions

  • the present invention relates to novel nucleic acid molecules which code for a protein with the biological activity of the tumor suppressor pl4 (ARF) and are characterized in that the nucleic acid sequence of the nucleic acid molecule has at least one mutation relative to the sequence coding for the native pl4 (ARF), which leads to a Resistance of the pl4 (ARF) variant encoded thereby leads to proteolytic degradation.
  • the present invention also relates to nucleic acid molecules which encode peptides which prevent its proteolytic degradation by binding to pI4 (ARF).
  • the present invention further provides the above pI4 (ARF) variant and peptides as well as the vectors containing the inventive nucleic acid molecules, these being suitable, for example, for gene therapy.
  • the conversion from normal cells to cancer cells takes place in several steps, which involve the mutation of cellular genes (proto-oncogenes and tumor suppressor genes) or the acquisition of viral oncogenes.
  • Cancer-relevant changes include the activation of proto-oncogenes by mutations, gene amplification, overexpression or chromosomal arrays, as well as the activation of tumor suppressor genes by mutations, for example deletions.
  • the tumor suppressor genes proved to be of particular interest, since they obviously offer new opportunities in cancer therapy and, moreover, their study allows a better understanding of the molecular mechanisms of cancer development.
  • p53 which encodes a transcription factor p53, the inactivation of which leads to the development of Li-Fraumeni syndrome, and which is found in the analysis of human tumors as has highlighted one of the most commonly mutated tumor suppressor genes (see, for example, Hollstein et al., Science 253 (1991), 49-53).
  • pl6 Two other tumor suppressors are pl6 (INK4A) (Serrano et al., Nature 366 (1993), 704-707) and pl4 / pl9 (ARF) (Quelle et al., Cell 83 (1995), 993-1000; Duro et al ., Oncogene 1_1 (1995), 21-29), which are involved in the arresting of cell growth and in tumor suppression and are encoded by the tumor suppressor locus INK4.
  • pl4 (ARF) corresponds to the human form of pl9 (ARF).
  • cancer therapy has mainly been carried out by cytostatic treatments of cancer patients with often relatively unspecific cytostatics or by radiation treatment.
  • leukaemias are also treated by bone marrow transplants.
  • the treatment of tumors with the help of cytostatics or radiation treatment has considerable disadvantages based on massive side effects, such as nausea, hair loss, leukopenia, organ damage (especially the liver and kidneys), etc. Bone marrow transplantation is associated with a great deal of effort and is only possible in individual cases , especially since there is usually the problem of finding a suitable donor.
  • the invention is essentially based on the technical problem of providing agents for cancer therapy which do not have the disadvantages of the previous therapy methods mentioned above, i.e. Above all, allow targeted therapy with no side effects.
  • cellular tumor suppressor protein pl9 (ARF) or pl4 (ARF) is a substrate for the proteasome. Furthermore, it has been found that the concentration of this protein is extremely low in a number of different tumor cell lines, although the corresponding mRNA is strongly expressed.
  • p53 lowers the expression of the gene encoding pl4 (ARF) by destabilizing the pl4 (ARF) protein and presumably also by downregulating transcription.
  • a complex control system comprising a p53 / MDM2 network is needed to maintain constant expression of p53 at a low level.
  • This network also responds to signals from other proteins.
  • the balance between MDM2 and p53 can be deregulated by pl4 (ARF).
  • p53 has the potential to destabilize the pl4 (ARF) protein. Control of the stability and function of p4 (ARF) by p53 could thus restore the balance between all three partner proteins.
  • the tumor suppressor function of pI4 can be achieved by the inventive procedures, which is therefore a new one Represent form of therapy, be restored.
  • a pl4 (ARF) variant can be produced which is resistant to proteolytic degradation
  • low-molecular peptides can be produced which bind to pl4 (ARF) and thus prevent its degradation and thus contribute to its stabilization.
  • These compounds can, for example, be expressed in the tumor cells by means of suitable vectors, as a result of which the tumor growth is stopped and / or programmed cell death (apoptosis) is triggered, as a result of which complete elimination of the tumor could be achieved.
  • one embodiment of the present invention relates to a nucleic acid molecule which encodes a protein with the biological activity of the tumor suppressor pl4 (ARF) and is characterized in that the nucleic acid sequence of the nucleic acid molecule has at least one mutation compared to the nucleic acid sequence coding for the native pl4 (ARF) leads to a resistance of the coded pl4 (ARF) variant to proteolytic degradation.
  • ARF tumor suppressor pl4
  • protein with the biological activity of the tumor suppressor pl4 refers to any protein which has at least one of the biological properties of pl4 (ARF) or pl9 (ARF), i.e. Tumor suppressor function, for example by promoting the breakdown of MDM2 (Pomerantz et al., Cell 92 (1998), pp. 713-723).
  • pl4 (ARF) variant refers to any form of pl4 (ARF) which is modified in relation to the native form in such a way that it is resistant to proteolytic degradation. This is preferably achieved by changing amino acid motifs which are recognition sites for proteases in such a way that on the one hand they are no longer recognized by the proteases and on the other hand the biological activity of the protein is essentially retained. This can be done by the targeted insertion of point mutations in the pI4ARF cDNA (Duro et al., Oncogene 11 (1995), pp. 21-29; see Fig. 2) by means of the polymerase chain reaction.
  • any desired mutation of the amino acid sequence of the resulting pl4ARF protein can be generated in a targeted manner.
  • These mutations will preferentially affect amino acids in the N-terminus of pARF14, since N-terminal sequences are of crucial importance for protein stability.
  • other mutations of the protein are also conceivable, which lead to resistance to proteasomal degradation.
  • the pl4 (ARF) variant according to the invention can have, in addition to these changes, further changes compared to the native form, ie, deletions, additions or exchanges of one or more amino acids and / or (a) modified amino acid (s) or changed compared to the native form Oligosaccharide side chains, whereby their biological activity is essentially retained, ie it has, for example, the properties described in the examples below.
  • the exchanges preferably include "conservative" exchanges of amino acid residues, ie exchanges for biologically similar residues, for example the substitution of a hydrophobic residue (for example isoleucine, valine, leucine, methionine) for another hydrophobic residue, or the substitution of one polar residue for another polar residue (e.g. arginine against lysine, glutamic acid against aspartic acid etc.).
  • Deletions can lead to the generation of molecules which are significantly smaller in size (fragments), ie which, for example, lack amino acids at the N or C terminus.
  • the above variants also relate to pI4 (ARF) variants which have a similar or better biological activity compared to the original form.
  • the term "resistance to proteolytic degradation” as used herein refers to the resistance achieved by at least one amino acid change that relates to the most effective protease (s), preferably all proteases of the proteasome, e.g. Serine / threonine and cysteine proteases as well as metalloproteases and acidic proteases (Goldberg, Science 268 (1995), pp. 522-523), or leads to a pl4 (ARF) variant which allows their presence in the cell in a concentration, that she can still act as a tumor suppressor.
  • s most effective protease
  • s preferably all proteases of the proteasome, e.g. Serine / threonine and cysteine proteases as well as metalloproteases and acidic proteases (Goldberg, Science 268 (1995), pp. 522-523), or leads to a pl4 (ARF) variant which allows their presence in the cell in a concentration, that she can still act as
  • the nucleic acid molecule according to the invention is furthermore characterized in that the pl4 (ARF) variant encoded thereby is not significantly impaired with regard to its interaction with p53
  • the present invention further relates to a nucleic acid molecule which encodes a peptide (aptamer) which binds to pl4 (ARF) in such a way that its proteolytic degradation is prevented but the interaction with p53 is not significantly impaired.
  • a peptide aptamer
  • Such peptides can also be used to prevent the proteolytic degradation of pI4 (ARF).
  • the person skilled in the art is able to design such peptide aptamers or peptides according to customary methods, for example the method described in Example 5 below, and to check their function.
  • Such peptides or peptide aptamers usually have a length of about 20 amino acids. However, shorter or longer fragments can also be active in the respective system.
  • peptide aptamers which have the desired properties can then be used as "lead compounds" for the derivation of suitable active substances, ie synthetic peptides which, for example, comprise essentially the same amino acid sequence as the peptide aptamer, but are protected against proteolytic degradation by modifications.
  • suitable active substances ie synthetic peptides which, for example, comprise essentially the same amino acid sequence as the peptide aptamer, but are protected against proteolytic degradation by modifications.
  • suitable active substances ie synthetic peptides which, for example, comprise essentially the same amino acid sequence as the peptide aptamer, but are protected against proteolytic degradation by modifications.
  • suitable active substances ie synthetic peptides which, for example, comprise essentially the same amino acid sequence as the peptide aptamer, but are protected against proteolytic degradation by modifications.
  • the processes for the preparation of peptides are known to the person skilled in the art (for example by means of Merryfield synthesis).
  • the nucleic acid molecules according to the invention can also be inserted into a vector.
  • the present invention also encompasses vectors containing these nucleic acid molecules.
  • the term "vector” refers to a plasmid (eg pUC18, pBR322, pBlueScript), a virus or another suitable vehicle.
  • the nucleic acid molecule according to the invention is functionally linked in the vector to regulatory elements which allow its expression in prokaryotic or eukaryotic host cells.
  • regulatory elements for example a promoter
  • such vectors typically contain an origin of replication and specific genes which allow the phenotypic selection of a transformed host cell.
  • the regulatory elements for expression in prokaryotes for example E.
  • coli include the lac, trp promoter or T7 promoter, and for expression in eukaryotes the AOX1 or GAL1 promoter in yeast, and the CMV, SV40 , RVS-40 promoter, CMV or SV40 enhancer for expression in animal cells.
  • suitable promoters are the metallothionein I and the polyhedrin promoter.
  • Suitable vectors include, for example, T7-based expression vectors for expression in bacteria (Rosenberg et al., Gene 56 (1987), 125, pMSXND for expression in mammalian cells (Lee and Nathans, J.Biol. Chem. 263 (1988), 3521, and baculovirus-derived vectors for expression in insect cells.
  • the nucleic acid sequence according to the invention is present in the vector in such a way that a fusion protein is encoded, which is a pl4 (ARF) variant or the peptide (aptamer) described above and a penetration sequence which is the inclusion in the desired target cell relieved.
  • a fusion protein which is a pl4 (ARF) variant or the peptide (aptamer) described above and a penetration sequence which is the inclusion in the desired target cell relieved.
  • penetration sequences can be obtained, for example, from the Drosophila Antennapedia protein (which contains the so-called "penetratin” motif) (Fahraeus et al., Current Biology 6 (1996), pp. 84-91) or the TAT protein from HIV- 1 (Vives et al., J.Biol.Chem., 222, (1997), 16010-16017) can be derived.
  • the vector containing the nucleic acid molecules according to the invention comes from a virus, for example from an adeno-associated virus
  • nucleic acid molecules according to the invention can also be transported to the target cells in the form of colloidal dispersions. These include, for example, liposomes or lipoplexes
  • the above vectors can also be used, for example, for ex vivo gene therapy, for example in the blood cells of patients with leukemia.
  • the present invention also relates to host cells containing the vectors described above.
  • host cells include bacteria, yeast, insect and animal cells, preferably mammalian cells. Methods for transforming these host cells, for phenotypically selecting transformants and for expressing the nucleic acid molecules according to the invention using the vectors described above are known in the art.
  • the present invention also relates to the pl4 (ARF) variant encoded by the above nucleic acid molecules or vectors containing them, or the peptide (aptamer) useful for stabilizing them before proteolytic degradation, these compounds preferably carrying at least one (further) modification, which protects against proteolytic degradation in the cell.
  • modifications are known to the person skilled in the art and include, for example, the modifications described above.
  • the most important modifications to protect peptides from degradation by cellular proteases are the attachment of sugar side chains to individual amino acids (N- or 0-glycosylations). Protection against the proteasomal degradation of pl4ARF is preferably provided by a targeted mutation of the N-terminus, since this is of crucial importance for protein stability.
  • the present invention also relates to a process for the production of the pl4 (ARF) variant or the peptide, comprising culturing the host cells described above under conditions which allow expression of the protein or peptide (preferably stable expression), and obtaining the protein or Culture peptides.
  • Suitable methods for the recombinant production of the protein or peptide are generally known (see, for example, Holmgren, Annu.Rev.Biochem. 54 (1985), 237; LaVallie et al., Bio / Technology 11 (1993), 187; Wong, Curr. Opin. Biotech. 6 (1995), 517; Romanos, Curr. Opin. Biotech .6 . (1995), 527; Williams et al. , Curr. Opin.
  • the present invention further relates to medicaments which contain the nucleic acids, vectors, proteins or peptides described above.
  • These drugs may also contain a pharmaceutically acceptable carrier.
  • Suitable carriers and the formulation of such medicaments are known to the person skilled in the art.
  • Suitable carriers include, for example, phosphate-buffered saline solutions, water, emulsions, for example oil / water emulsions, wetting agents, sterile solutions, etc.
  • the medicaments can be administered orally or parenterally. Methods for parenteral administration include topical, intra-arterial (e.g. direct to the tumor), intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, or intranasal.
  • the appropriate dosage is determined by the attending physician and depends on various factors, for example the age, gender, weight of the patient, the stage of a tumor, the type of administration, etc.
  • the medicament according to the invention is preferably used for the prevention or treatment of cancer. These include, for example, leukemia, melanomas, gliomas and tumors of the bladder.
  • the medicament according to the invention is particularly preferably used for the treatment of leukemia.
  • the medicament can be used in gene therapy, and the methods or vectors described above can be used to determine the nucleic acids according to the invention.
  • Nucleic acid molecules encoded protein or peptide are administered directly, so as to restore tumor suppressor activity in cells that do not have a sufficient concentration of pl4 (ARF).
  • the medicament according to the invention is preferably used when suitable diagnostic methods, for example methods based on the analyzes described in the examples below, provide evidence that biologically active pI4 (ARF) is not present in the patient concerned or is present in insufficient quantities is available.
  • the present invention relates to the use of a nucleic acid molecule, vector, protein or peptide according to the invention for the manufacture of a medicament for the treatment or prevention of cancer, which is preferably leukemia.
  • Hpl9 ARF used in FIG. 1 is synonymous with pl4 (ARF), i.e. represents the human homologue of the mouse protein commonly designated pl9 (ARF).
  • SAOS cells and the subclone X4-4 expressing p53 were grown as indicated at 32 ° C and 37 ° C, respectively.
  • Metabolic labeling and immunoprecipitation were performed as in DellaValle et al. , Oncogene 15 (1996), 2949-2959, using rabbit anti-pl4 (ARF) antiserum or the pre-Iramun serum as indicated.
  • B, C stabilization of pl4 (ARF) by the proteasome inhibitor N-acetyl-eu-Leu-norleucinal (LLnL)
  • B C33A cells were transfected with an expression vector for pl4 (ARF) which was under the control of the CMV promoter and incubated for 12 hours with the protease inhibitor (50 ⁇ , M) indicated in each case.
  • Oncogene 15 (1996), 2949-2959, described using rabbit anti-pl4 (ARF) antiserum and the monoclonal anti-p53 antibody DO-1 (Santa Cruz, Heidelberg).
  • the lower section of the figure shows a longer exposure of the pl4 (ARF) immunoblot.
  • C33A cells transfected with pl4 (ARF) were treated for 12 hours with different concentrations of N-acetyl-Leu-Leu-Norleucinal (LLnL; Sigma, Deisenhofen) as indicated and analyzed as in (B).
  • LnL N-acetyl-Leu-Leu-Norleucinal
  • C33A cells were transfected together with a pl4 (ARF) expression vector and different amounts of a p53 expression vector.
  • the cell extracts were analyzed by means of immunoblots for pI4 (ARF) and p53 as in (B). The ratio between the two expression vectors varied from 1: 1 to 1:60.
  • C33A cells were transfected with the expression vectors for pl4 (ARF) and p53 in the ratio 1: 1, treated with DMSO or 50 ⁇ M of the protease inhibitors indicated and analyzed by Western blot as in (B).
  • p53 induces the expression of the MDM2 gene and MDM2 triggers the degradation of p53 by the proteasome, while pl4 (ARF) Destabilization of p53 by MDM2 inhibits (black lines) (Pomerantz et al., Cell 92 (1998), 3926-3931). In the present invention it is shown that p53 also down-regulates pl4 (ARF) (dashed line)
  • the cell lines U-20S (ATCC, Rockville, Md, number: HTB 96), Saos-2 (ATCC, Rockville, Md., Number: HTB 85) and those by transfection of the Xenopus laevis p53 cDNA (Bessard et al., Oncogene 16, (1998), pp. 883-890) cell line X4-4 produced in Saos-2 were in Dubecco's Modified Eagle Medium, supplemented with 100 U / ml penicillin, 0.1 rag / ml streptomycin, 2 mM glutamine and 10% fetal calf serum (FCS). The cultivation took place in coated cell culture dishes or bottles in an incubator at 37 ° C. in a water-saturated atmosphere and 5% CO 2 gassing. The cells were passaged every two days.
  • the expression plasmids for pl4ARF were produced by inserting the cDNA of pl4ARF into the CMV promoter-driven vector pX (Pagano et al., EMBO J. 11, pp. 961-971 (1992)).
  • the yeast vectors for the expression of the peptide aptamer bank were produced by inserting this bank into the plasmid pJM-1 (Colas et al., Nature 380 (1996), pp. 548-550).
  • the cells were transfected according to the protocol of
  • IP buffer 50 mM HEPES-NaOH, pH 7.0; 150 mM NaCl, 0.2 mM PMSF, 0.1 mM Na 3 V0 4 , 10 mM ⁇ -glycerophosphate, 10 ⁇ g / ml Aprotinin, 1 mM NaF, 0.1% NP40
  • the lysates were first pre-cleaned with 50 ⁇ l protein A or protein G agarose equilibrated in IP buffer for 1 hour at 4 ° C.
  • Example 2 p53 regulates the pI4 (ARF) level
  • p53-deficient SAOS-2 cells were labeled with an expression vector for Xl-p53 (Bessard et al., Oncogene 16 (1998), pp. 883-890) stably transfected, whereby the cell line X4-4 was obtained.
  • SAOS-2 and X4-4 cells were grown at 37 ° C. At this temperature, Xl-p53 is inactive.
  • Example 3 p53 triggers the proteolytic degradation of pl4 (ARF) by the
  • pl4 ARF
  • ARF pI4
  • U-20S cells ATCC, Rockville, Md.
  • CMV promoter which is the direct detection of the transfected protein allowed by Western blot; see FIG. 1B, lane 2
  • transiently infected and incubated with the protease inhibitor LLnL during the transfection experiment There was a significant increase in transfected pl4 (ARF) protein, while the unrelated protease inhibitors LLM and E46 had no effect.
  • the stabilization of pI4 (ARF) depends on the LLnL dose used (FIG. IC).
  • pl4 (ARF) is subject to degradation by the proteasome.
  • Treatment with LLnL leads to a significant loss of pl4 (ARF) protein in both pl4 (ARF) -expressing and non-transfected U-20S cells (FIG. 1B).
  • the common expression of p53 with pl4 (ARF) in U-20S cells leads to a dose-dependent elimination of pl4 (ARF) (FIG. 1D), while the p4 (ARF) mRNA level is not influenced by p53. Since the elimination of the pl4 (ARF) protein in the presence of p53 can be inhibited by the proteasome inhibitor LLnL (FIG. 1E), it can be assumed that p53 triggers the proteolytic degradation of pl4 (ARF) by the proteasome.
  • a nucleic acid sequence was isolated which encodes a pl4 (ARF) variant which is resistant to proteolytic degradation is the proteasome, but can still stabilize p53.
  • point mutations were inserted in the section of the p4ARF cDNA coding for the N-terminus (Duro et al., Oncogene 11 (1995), pp. 21-29) by means of the polymerase chain reaction (PCR). This PCR was carried out with primer oligonucleotides, each in a nucleotide of the wild-type sequence differed.
  • Peptide aptamers are being developed which can bind to the pl4 (ARF) protein so specifically that proteolytic degradation is prevented, but the interaction with the p53 protein is not disturbed.
  • the pl4 (ARF) cDNA (Duro et al., Oncogene 11 (1995), pp. 21-29) is inserted into the yeast expression vector pEG202 (Gyuris et al., Cell 75 (1993), pp. 791-803 ) which allows the peptides from a peptide bank with randomly generated sequences to be used in yeast cells using the so-called "two hybrid" system (Gyuris et al., Cell 75 (1993), pp.
  • peptide aptamers usually consist of about 20 amino acids, which are expressed in the context of the bacterial thioredoxin A protein (TrxA). However, shorter or longer peptides are also possible, and expression can also be in the context of another "Framework protein" as TrxA.
  • TrxA bacterial thioredoxin A protein
  • the interaction of such a peptide with the pl4ARF protein is identified in the "two-hybrid system" by the fact that only in the yeast cells in which such an interaction takes place is a reporter gene expressed which allows the cells to reach nutrient media grow that lack an essential amino acid.
  • DNA sequences encoding peptide aptamers are then cloned into a mammalian expression vector under the control of a CMV promoter, eg pCDNA3, pCI or the like, and expressed in mammalian cells, eg U-20S, together with the pl4 (ARF) protein.
  • a CMV promoter eg pCDNA3, pCI or the like
  • the influence of certain peptide aptamers on the metabolic stability of the pl4 (ARF) protein can be investigated by a Western blot analysis with extracts from transfected cells.
  • Peptide aptamers are sought that inhibit the proteolytic degradation of pI4 (ARF) but do not interfere with its interaction with p53.
  • the p53 binding is examined by the joint expression of a p53 expression vector and subsequent immunoprecipitation by means of commercially available anti-p53 antibodies. If pl4ARF is in a complex with p53 in these cells, it is coprecipitated by precipitation with anti-p53 antibodies and can be detected by Western blot analysis. Resistance to proteases is tested by transfecting the pl4ARF gene into cells either alone or together with the sequence for a specific binding peptide aptamer. These are then metabolically labeled by adding radiolabelled methionine and cysteine. The proteins synthesized in a defined period of time (approx. 30 minutes) can be visualized by autoradiography after immunoprecipitation and electrophoretic separation. A larger amount of remaining pl4ARF protein in the presence of a peptide aamer compared to that expressed in control cells indicates an increased resistance to proteolytic degradation.

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Abstract

L'invention concerne un variant du suppresseur de tumeurs p14 (ARF) qui présente une résistance vis-à-vis de la décomposition protéolytique, ainsi qu'une molécule d'acide nucléique codant pour ce variant. Sont également décrits des peptides qui empêchent la décomposition protéolytique du suppresseur p14 (ARF) en se liant à celui-ci, ainsi que des molécules d'acide nucléique codant pour ces peptides. Sont également décrits les vecteurs contenant lesdites molécules d'acide nucléique, lesquels peuvent être utilisés, par exemple, pour la thérapie génique de tumeurs. L'invention concerne enfin des médicaments qui contiennent les molécules d'acide nucléique ou les vecteurs susmentionnés, ou bien les protéines ou peptides codés par ces molécules ou vecteurs, de préférence pour le traitement de la leucémie.
PCT/DE2000/004257 1999-11-30 2000-11-29 Suppresseur de tumeurs p14 (arf) resistant a la protease Ceased WO2001040300A2 (fr)

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DE1999157689 DE19957689A1 (de) 1999-11-30 1999-11-30 Proteaseresistenter Tumorsuppressor p14(ARF)
DE19957689.0 1999-11-30

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Cited By (6)

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EP1387173A1 (fr) * 2002-07-29 2004-02-04 MTM Laboratories AG Procédé servant à améliorer le diagnostic de lésions cervicales fondé sur la détection de produits géniques INK4a
EP1386929A1 (fr) * 2002-07-29 2004-02-04 MTM Laboratories AG Peptides associés à des tumeurs et leurs utilisations
WO2004013631A3 (fr) * 2002-07-29 2004-04-01 Mtm Lab Ag Compositions et methodes de diagnostic et de traitement du cancer
US8043819B2 (en) 2002-04-09 2011-10-25 Mtm Laboratories, Ag Method for discrimination of metaplasias from neoplastic or preneoplastic lesions
US8367353B2 (en) 2002-10-28 2013-02-05 Roche Mtm Laboratories Ag Method for improved diagnosis of dysplasias
EP3420348A4 (fr) * 2016-02-22 2019-10-02 Healthspandx Procédés de prévention ou de réduction d'une insuffisance rénale aiguë

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Publication number Priority date Publication date Assignee Title
US8043819B2 (en) 2002-04-09 2011-10-25 Mtm Laboratories, Ag Method for discrimination of metaplasias from neoplastic or preneoplastic lesions
US10246751B2 (en) 2002-04-09 2019-04-02 Ventana Medical Systems, Inc. Method for discrimination of metaplasias from neoplastic or preneoplastic lesions
EP1387173A1 (fr) * 2002-07-29 2004-02-04 MTM Laboratories AG Procédé servant à améliorer le diagnostic de lésions cervicales fondé sur la détection de produits géniques INK4a
EP1386929A1 (fr) * 2002-07-29 2004-02-04 MTM Laboratories AG Peptides associés à des tumeurs et leurs utilisations
WO2004013631A3 (fr) * 2002-07-29 2004-04-01 Mtm Lab Ag Compositions et methodes de diagnostic et de traitement du cancer
US8367353B2 (en) 2002-10-28 2013-02-05 Roche Mtm Laboratories Ag Method for improved diagnosis of dysplasias
US8975036B2 (en) 2002-10-28 2015-03-10 Ventana Medical Systems, Inc. Methods for improved diagnosis of dysplasias
US9528159B2 (en) 2002-10-28 2016-12-27 Ventana Medical Systems, Inc. Method for improved diagnosis of dysplasias
EP3420348A4 (fr) * 2016-02-22 2019-10-02 Healthspandx Procédés de prévention ou de réduction d'une insuffisance rénale aiguë

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