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WO2008015383A2 - Activité anticancéreuse des protéines bmp-9 et bmp-10 et leur utilisation dans les traitements du cancer - Google Patents

Activité anticancéreuse des protéines bmp-9 et bmp-10 et leur utilisation dans les traitements du cancer Download PDF

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WO2008015383A2
WO2008015383A2 PCT/GB2007/002755 GB2007002755W WO2008015383A2 WO 2008015383 A2 WO2008015383 A2 WO 2008015383A2 GB 2007002755 W GB2007002755 W GB 2007002755W WO 2008015383 A2 WO2008015383 A2 WO 2008015383A2
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bmp
cells
prostate cancer
polypeptide
breast
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WO2008015383A8 (fr
WO2008015383A3 (fr
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Wen Guo Jiang
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University College Cardiff Consultants Ltd
Cardiff University
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University College Cardiff Consultants Ltd
Cardiff University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1875—Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to two novel agents, BMP-9/BMP-10, for treating breast or prostate cancer; vectors adapted to transform or transfect breast or prostate tissue which vectors are designed to express either, or both, BMP-9 and BMP-10; cell lines which have been transformed or transfected so as to express native or recombinant BMP-9 and/or BMP-10; a method for the recombinant production of BMP-9 and/or BMP-10; and methods for treating breast or prostate cancer comprising exposing breast or prostate cancer tissue to BMP-9 and/or BMP-10.
  • Bone morphgenesis proteins are a protein family that belongs to the TGF ⁇ (transforming growth factor beta) superfamily. BMPs are widely involved in the regulation of cell functions including cell growth and apopotosis. More than 20 BMPs have been reported so far. The proteins are also important in embryo development and tissue regeneration. Some of the BMPs are also known to be important players in cancer. For example, BMP-2, BMP-6 and BMP-7 have been shown to be aberrantly expressed in human cancers and that raised levels of the proteins either in the circulation or in tumours are associated with progression and spread of cancer.
  • TGF ⁇ transforming growth factor beta
  • BMPs exert their effects through a heteromeric receptor complex, which consists of two types of serine-threonine kinase transmembrane receptors.
  • the Type-I receptors include BMP receptor type IA (BMPR-IA) 1 type IB (BMPR-IB), Activin receptor-like kinase-1 (ALK-1), ALK-4, ALK-5 and activin A receptor type I (ActRI).
  • the Type-ll receptors include BMP receptor type Il (BMPR-II), activin A receptor type HA (ActRII) and activin A receptor type MB (ActRIIB). Upon binding to BMP ligands, the Type-ll receptors then phosphorylates the Type-I receptors.
  • R-Smads pathway-restricted Smads
  • Smadsi pathway-restricted Smads
  • BMP-9 and BMP-10 have a very strong anti-cancer functions. This is in clear contrast to other BMPs of which high levels are most associated with pro-cancer actions.
  • BMP-9 otherwise known as GDF2 (Growth and Differentiation Factor-2) was first found in liver cells (Celeste et al 1994) and the major source is Kupffer, Stellate and endothelial cells (Miller et al 2000). In embryos, BMP-9 was highly expressed in neural tissues and therefore has been indicated in the development of nervous system. In bones, BMP-9 was able to induce differentiation of osteoclast. BMP-10, discovered in 1999 (Neuhaus et al 1999), is known to be expressed at good levels in heart and lung and has also been indicated in embryo development such as heart. Statements of Invention
  • Our invention concerns the role of BMP-9 and/or BMP-10 in breast or prostate cancer and more particularly the inhibitory role of BMP-9 and/or BMP- 10 in breast or prostate cancer.
  • an anti- cancer agent for treating breast or prostate cancer comprising a nucleic acid molecule encoding BMP-9, as shown in Figure 9, or a homologue thereof, or a nucleic acid molecule that hybridises to the sequence shown in Figure 9 under stringent conditions.
  • an anticancer agent for treating breast or prostate cancer comprising a nucleic acid molecule encoding BMP-10, as shown in Figure 10, or a homologue thereof, or a nucleic acid molecule that hybridises to the sequence shown in Figure 10 under stringent conditions.
  • Reference herein to stringent conditions includes reference to either increasing the temperature of incubation to above 25 0 C and more preferably above 5O 0 C and more preferably still up to 65 0 C and/or washing the annealed molecules using a salt solution having an ionic strength of 1.0N sodium chloride - 0.02N sodium chloride, and most preferably 0.5N sodium chloride - 0.02N sodium chloride and more preferably still 0.1 N sodium chloride to 0.02N sodium chloride.
  • polypeptide that has anti-cancer activity in treating breast or prostate cancer comprising the polypeptide BMP-9 as shown in Figure 9, or a homologue thereof, or a fragment thereof which has BMP-9 anti-cancer activity.
  • a polypeptide that has anti-cancer activity in treating breast or prostate cancer comprising the polypeptide BMP-10 as shown in Figure 10, or a homologue thereof, or a fragment thereof which has BMP-10 anti-cancer activity.
  • a nucleic acid molecule encoding BMP-9 and/or BMP-10 for treating breast or prostate cancer.
  • BMP-9 or BMP-10 polypeptide for treating breast or prostate cancer.
  • a medicament comprising either the nucleic acid molecule encoding BMP-9 and/or BMP-10, and/or the corresponding BMP-9 and/or BMP-10 polypeptide.
  • the medicament of the invention is most suitable for treating breast or prostate cancer.
  • the medicament is formulated with a suitable excipient, carrier or emollient.
  • a method for treating breast or prostate cancer comprising administering to an individual to be treated a medically effective amount of BMP-9 and/or BMP-10.
  • a method for treating breast or prostate cancer comprising increasing cellular levels of either exogenous or endogenous BMP-9 and/or BMP-10 in breast or prostate cells.
  • Reference herein to the exogenous increase in cellular levels of BMP-9 and/or BMP-10 includes reference to: introducing into breast and/or prostate cells either externally manufactured BMP-9 and/or BMP-10 polypeptide; or introducing means for increasing the said cells production of BMP-9 and/or BMP-10 by, for example, introducing a copy of, or increasing the number of copies of, the BMP-9 and/or BMP-10 gene.
  • Reference herein to the endogenous increase in cellular levels of BMP-9 and/or BMP-10 polypeptide includes introducing into breast and/or prostate cells means that enables the breast or prostate cells to increase native production of BMP-9 and/or BMP-10 polypeptide, for example, suitable means include supplementing the cells' manufacturing machinery to provide for the increased production of BMP-9 and/or BMP-10 such as by increasing the effectiveness or activity of the promoter(s) controlling the expression of BMP-9 and/or BMP-10 gene whereby the overall production of BMP-9 and/or BMP-10 is increased, or, additionally or alternatively, supplementing the cells' other cellular protein production pathways whereby the amount of BMP-9 and/or BMP-10 produced is increased.
  • suitable means include supplementing the cells' manufacturing machinery to provide for the increased production of BMP-9 and/or BMP-10 such as by increasing the effectiveness or activity of the promoter(s) controlling the expression of BMP-9 and/or BMP-10 gene whereby the overall production of BMP-9 and/or BMP-10 is increased, or, additionally or
  • a vector adapted to transfect or transform breast or prostate cells wherein said vector includes:
  • At least one over-expressing or constitutively active promoter which is either coupled to the BMP-9 and/or BMP-10 gene of part 1. and/or which is designed for insertion into a genome upstream of the native BMP-9 and/or BMP- 10 gene of said breast or prostate cells; whereby transfection or transformation of said cells with said vector results in the enhanced expression of BMP-9 and/or BMP-10.
  • a cell line which has been transfected or transformed with a vector encoding at least one copy of the gene for BMP-9 and/or BMP-10 or as described herein.
  • a 3T3 cell line or a PC-3 cell line, or a DU145 cell line, or a MDA231 cell line or a T24 cell line which has been transfected or transformed so as to recombinantly express BMP-9 and/or BMP-10.
  • BMP-9 and/or BMP-10 which has been manufactured by a host cell that has been transfected or transformed with a vector described herein.
  • a method for manufacturing recombinant BMP-9 and/or BMP-10 which method comprises:
  • said vector further includes a suitable secretion signal whereby once BMP-9 and/or BMP-10 polypeptide has been produced it is processed through the cell's machinery for secretion and therefore the secreted polypeptide can be harvested from the extra cellular medium.
  • BMP-9 and/or BMP-10 to treat breast or prostate cancer.
  • BMP-9 and/or BMP-10 in the manufacture of a medicament to treat breast or prostate cancer.
  • a BMP-9 or BMP-10 receptor or downstream signalling molecule such as Par-4 or Smad- 1 , or Smad-5 or Smad-8, or an agonist or enhancer of any of the afore molecules for use in treating breast or prostate cancer.
  • FIG 1 shows Figure-1. Staining of BMP-9 in mammary (left two panels) and prostate (right two panels) tissues (normal (left) and tumour (right) tissues are shown in the respective tissue type). Normal epithelial cells stained positively for BMP-9. The protein staining, however, was lost in tumour cells (right).
  • FIG 2 shows Staining of BMP-10 in mammary (left two panels) and prostate (right two panels) tissues (normal (left) and tumour (right) tissues are shown in the respective tissue type). Normal epithelial cells stained positively for BMP-10. The protein staining, however, was lost in tumour cells (right).
  • the top panel shows correlation between BMP transcripts and prognosis, in which NPI-1 indicating patients with good prognosis, NPI-2 with moderate and NPI-3 with a poor prognosis. Patients with a poor prognosis had significantly lower levels of BMP-10;
  • Figure 4 shows expression of BMP-9 and BMP-10 in cancer cells, as revealed by RT-PCR.
  • FIG. 5 shows the construction and generation of BMP-9 (A and B) and BMP-10 (C and D) expression vector.
  • a and C are selected from transformed E. CoIi.
  • C and D are purified, respective, plasmids;
  • Figure 6 shows the establishment of BMP-9 in PC-3 prostate cancer cells (top) and BMP-9 and BMP-10 (bottom) in MDA MB 231 breast cancer cells. Shown are mRNA from PC-3 cells (top), and mRNA from MDA MB 231 cells (bottom);
  • Figure 6A shows the amount of BMP-9 protein produced in PC-3 cells
  • Figure 7 shows protein expression in transfected cancer and other mammalian lines.
  • Successful cell lines were established in 3T3 (fibroblast) for BMP-9; 3T3 (fibroblast), PC-3 (prostate) and T24 (bladder) cells for BMP-10, (HECV is an endothelial cell line and DU14 is a prostate cell line);
  • Figure 8 shows over-expression of BMP-9 and BMP-10 in cancer cells (MDA MB-231 breast cancer, PC-3 prostate cancer and DU145 prostate cancer) resulted in a slower growth and reduced invasion;
  • Figure 9 shows the impact of BMP-9 on the ability of prostate cancer cells to adhere to the Extracellular matrix
  • Figure 10 shows the effect of BMP-9 over-expression on cellular motility using an in vitro migration assay
  • FIG 11 shows BMP-9 induces apoptosis in PC-3 cells via Par-4.
  • A overexpression of BMP-9 induces apoptosis in PC-3 cells using a flow cytometry.
  • the percentage of the apoptotic cells in pc-3 BMP"9exp cells is 29.4% (top), which is remarkably higher than the 0.45% of pc-3 pEF/His cells (middle) and 5.00% of PC-S ⁇ (bottom).
  • B top panel shows an increase of Par-4 transcripts in PC-3BMP-9exp cells using RT-PCR 1 while the lower panel of B shows the elevated level of Par-4 protein in PC-3 by over-expression of BMP-9 using Western blot analysis.
  • Figure 12 shows Both BMPR-1 B and BMPR-2 mediate inhibitory effects on cell growth of prostate cancer cells.
  • A knockdown of BMPR-IB and BMPR-II mRNA in PC-3 cells by the ribozyme transgenes was verified using RT-PCR.
  • the gene transcripts of BMPR-IB were diminished in the pc-3 ⁇ BMPR IB cells by the ribozyme transgenes (top).
  • the PCR reactions were performed 30 cycles.
  • the mRNA level of BMPR-II was also markedly reduced in the pc-3 ⁇ BMPR" ", compared with the PC-S ⁇ and pc-3 pEF/His (bottom).
  • the ribozyme transgenes reduced the protein production of BMPR-IB and BMPR-II in PC-3 cells using western blot analysis.
  • the protein production of BMPR-IB was eliminated in the pC 3 ⁇ BMPR-IB cel
  • the cell growth rate at day 4 is 239.3% ⁇ 30.7% of PC-3 m cells and 209.9% ⁇ 26.9% of pc _ 3 pEF/His ce
  • The' growth rate was calculated as a percentage using the absorbance of day 1 as a baseline.
  • Figure 13 shows BMP-9 signals through a Smad-dependent pathway in PC-3 cells.
  • A effects of rh-BMP-9 on in vitro cell growth of PC-3 cells was examined using the in vitro cell growth assay. The cell growth of PC-3 cells were reduced by the exposure of rh-BMP-9 at 50ng/ml. Error bars are standard deviations. * p ⁇ 0.05 vs control.
  • B rh-BMP-9 (20ng/ml) induced apoptosis in PC- 3 cells using flow cytometry. The percentage of cells in exposure to rh-BMP-9 undergoing apoptosis is 16.88%, which is higher than the control (7.73%).
  • C BMP-9 signal through Smad dependent pathway in PC-3 cells.
  • BMPR-IB top
  • BMPR-II middle
  • D lmmunofluorescent staining of phosphorylated Smad-1 by BMP-9 in PC-3 cells.
  • Figure 14 shows the nucleic acid sequence and corresponding amino acid sequence of BMP-9.
  • Figure 15 shows the nucleic acid sequence and the corresponding amino acid sequence of BMP-10. Materials and Methods Human tumour tissues
  • PC-3 ECACC, European Collection of Animal Cell Culture, Salisbury, UK
  • DU-145, LNCapFGC, CA-HPV10 and PZ-HPV-7 ATCC, American Type Culture Collection, Mannasas, Virginia
  • Polyclonal goat anti-BMP-9, monoclonal mouse anti-actin, polyclonal goat anti-phospho Smad 1 and monoclonal mouse anti-Par-4 were obtained from Santa Cruz Biotechnology (Santa Cruz, California, USA).
  • Other reagents or kits were obtained from Sigma- Aldrich, Poole, United Kingdom. Immuno-histochemical Staining of Breast and prostate tissue sections
  • Frozen sections of normal and tumours were cut at a thickness of 6 ⁇ m using a cryostat.
  • the sections were mounted on super frost plus microscope slides, air-dried and then fixed in a mixture of 50% Acetone and 50% methanol.
  • the sections were then placed in "Optimax" wash buffer for 5 -10 minutes to rehydrate.
  • Sections were incubated for 20 minutes in a 0.6% BSA blocking solution and probed with the primary antibodies. Following extensive washings, sections were incubated for 30 minutes in the secondary biotinylated antibody (Multilink Swine anti-goat/mouse/rabbit immunoglobulin, Dako Inc.). Following washings, the Avidin Biotin Complex (Vector Laboratories) was then applied to the sections, followed by extensive washing steps.
  • Diamino benzidine chromogen (Vector Labs) was then added to the sections, and incubated in the dark for 5 minutes. Sections were then counter stained in Gill's Haematoxylin and dehydrated in ascending grades of methanol before clearing in xylene and mounting under a cover slip.
  • Full length cDNA for the respective BMP were cleaned by gel separated and extraction, T-A cloned into an mammalian expression vector, pEF6/TOPO, which were subsequently used to transform the TOPO10 E. CoI.
  • Discrete bacterial colonies were screened for the presence of the coding sequence and direction of the inserts. Positive colonies were isolated, amplified, and plasmids subsequently extracted using a plasmid extraction kit. Following verification of the plasmids, purified plasmids were used to transfect breast and prostate cancer cell lines, followed by selection with blastidicin containing medium.
  • the first strand cDNA was synthesized from RNA isolated from normal human tissues using a DuraScriptTM RT-PCR kit s. PCR was then used to amplify the coding sequence of full-length human BMP-9 using the Extensor Hi-Fidelity PCR master mix (ABgene Ltd., Epsom, UK). The sequences of primers are shown in Table 3.
  • the verified BMP-9 insert was cloned into a mammalian expression plasmid vector (pEF/His TOPO TA plasmid vector, Invitrogen, Inc., Paisley, UK). The recombinant plasmid vectors were transformed into chemically competent TOP10 E. CoIi (Invitrogen, Inc., Paisley, UK), and the colonies were then analyzed. Colonies carrying correct recombinant plasmids were amplified, and the BMP-9 recombinant plasmids were then extracted
  • Anti-human BMPR-IB and anti-human BMPR-II hammerhead ribozymes targeting were designed based on the secondary structure of the respective gene generated using Zuker's RNA mFold program (Zuker et al 2003).
  • the ribozymes were individually cloned into a mammalian expression pEF6/V5-His- TOPO plasmid vector (Invitrogen Ltd., Paisley, UK). Ribozyme transgenes and control plasmid vectors were then transfected into PC-3 cells respectively. After up to 3 weeks selection using blasticidin, the transfectants were used in the current study.
  • the protein concentration in cell lysates were determined using the DC Protein Assay kit (BIO-RAD) and an ELx ⁇ OO spectrophotometer (BIO-TEKTM). Equal amount of proteins were separated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE), and blotted onto nitrocellulose sheets. Proteins were then probed with the anti-BMP-9/10 antibody and peroxidase-conjugated secondary antibodies. Protein bands were visualized using the SupersignalTM West Dura system (Pierce Biotechnology, Inc., Rockford, IL 1 USA), and photographed using a UVITech imager (UVITech, Inc., Cambridge, UK).
  • the plasmids thus generated were referred to as pEF6/BMP9Exp and pEF6/BMP10Exp.
  • the cells generated were referred to as
  • the BMP modified cancer cells were seeded in a 96 well plate at a density of 7,000 cells/well, and incubated at 37°C for up to 5 days. Cells were fixed and stained with Crystal violet. Absorbance was measured on a multiplate reader.
  • Transwell chambers equipped with a 6.5mm diameter polycarbonate filter insert (pore size 8 ⁇ m)(Becton Dickinson, Labware, Oxford, UK) 1 were pre- coated with 50 ⁇ g/insert of solubilised tissue basement membrane, Matrigel (Collaborative Research Products, Bedford, Massachusetts, USA). 40,000 cancer cells were seeded into each insert and allowed to invade for 3 days. Following incubation, cells that had invaded through the basement membrane were fixed (4% formaldehyde), and then stained with crystal violet. For analysis, the cells were counted in 10 fields/insert (x40 magnification); to determine the mean number of invaded cancer cells.
  • Cells were plated into a 96-well plate (2,500cells/well). Cell growth was assessed after a period of incubation (upto 5 days). Crystal violet was used to stain cells, and absorbance was determined at a wavelength of 540nm using a spectrophotometer (BIO-TEK, Elx800, UK).
  • Transwell inserts with 8 ⁇ m pore size were coated with 50 ⁇ g Matrigel (BD MatrigelTM Basement Membrane Matrix) and air dried. The Matrigel was rehydrated before use. 20,000 cells were added to each well. After 96 hours cells that had migrated through the matrix to the other side of the insert were fixed in 4% formalin, stained with 0.5% (weight/volume) crystal violet and counted under a microscope.
  • Matrigel BD MatrigelTM Basement Membrane Matrix
  • 50,000 Cells were seeded into each well of a chamber slide and allowed to reach near confluence. The layer of cells was then scraped with a fine gauge needle. The movement of cells to close the wound was recorded using a time lapse video recorder and analyzed using the Optimas 6.0 motion analysis.
  • PC-3 cells were seeded on glass chamber slide in a duplicated manner. Following 2 hours of incubation in serum free DMEM the cells were exposed to rh-BMP-9 (20ng), or serum free medium alone for 1 hour.
  • the immunofluorescent staining of phosphorylated Smad-1 was performed using anti-Phosph-Smad1 antibody is 1 :50 from original, and the TRITC labelled anti-goat IgG was used at 1 :200, and then photographed.
  • cancer cell lines had different pattern of expression of both BMP-9 and BMP-10.
  • BMP-9 and BMP1- 10 BMP-9 and BMP-10 were amplified from normal human cDNA bank and cloned into a mammalian expression vector. Following screening ( Figure 5 A and C), plasmids were extracted and purified ( Figure-5 B and D) and used for electroporation.
  • BMP-9 and BMP-10 over-expression resulted in death/senescence of cancer cells as well as CHO cells It was during the selection stage that we realised that over-expressing BMP-9 in CHO and PC-3 (prostate) and DU-145 (breast) results in cells rapidly developing cell senescence, and so we were unable to establish stably transfected cells. This suggests that BMP-9 triggers the cancer cells to terminal development, which eventually lead to death.
  • Figure-8 shows the growth of cancer cells that have stable integration for BMP-9 and BMP-10. These cells were selected under extreme conditions, as most cells were dead in most experiments. ⁇
  • pc3 BMP9exp cells showed a significantly reduced cellular migration, compared with the controls. A remarkable reduction of distance migrated was seen in these cells, 60 minutes after wounding. The average distance migrated over 90 minutes for pc-3 BMP'9exp cells was 17.22 ⁇ 3.61 ⁇ m, p ⁇ 0.05 vs both PC- ⁇ (32.47 ⁇ 2.98 ⁇ m) and pc-3 pEF/His (30.26 ⁇ 2.43 ⁇ m), as shown in Fig. 10.
  • Prostate apoptosis response-4 (Par-4) has been demonstrated as being up-regulated in androgen-independent prostate cancer cells which were induced to undergo apoptosis (28-30).
  • the PC-3 cells used in this study is androgen insensitive, and derived from bone metastasis of prostate cancer. Therefore, we hypothesised that Par-4 may be involved in the apoptosis induced by BMP-9.
  • FIG. 11B shows an up-regulation in the mRNA level of Par-4 in p C _ 3 BMP - 9e x p ce
  • An elevated protein level of Par-4 was shown in the pc-3 BMP"9exp cells (Fig. 11 B-lower panel).
  • the immunocytochemical staining also demonstrated an increased Par-4 protein in pc-3 BMP"9exp cells which appeared to be more condensed in the nuclei of the cells (Fig. 11C). This phenomenon is further elucidated in the immunofluorescent staining of Par-4 (Fig. 11 D).
  • Both BMP-9 and BMP-10 are expressed at lower levels in breast and prostate tumour tissues compared with normal tissues. The levels are particularly low in aggressive tumours. This suggests that loss of both BMPs are associated with an aggressive clinical condition in clinical cancer.
  • the study has further shown that over-expressing BMP-9 and BMP-10 in cancer cells lead to unexpected slower growth; reduced migration, motility, invasion and adhesion; and finally senescence and apoptosis in cancer cells.
  • Apoptosis is the key event for physiological growth control and regulation of tissue homeostasis.
  • BMP-9 inhibited in vitro cell growth of prostate cancer cells. This was not related to necrotic cell death, as observed during the study. This has lead us to discover that apoptosis is potentially the underlying mechanism for the reduced rate of cell growth induced by BMP9.
  • BMP-9 proapoptotic protein
  • Par-4 is a proapoptotic protein, which was originally identified in prostate cancer cells undergoing apoptosis in response to ionomycin.
  • Par-4 is involved in the apoptosis induced by tumor necrosis factor (TNF), doxorubicin, et ⁇ poside, UV irradiation, growth factor deprivation, and ionizing radiation.
  • TNF tumor necrosis factor
  • doxorubicin doxorubicin
  • et ⁇ poside UV irradiation
  • growth factor deprivation growth factor deprivation
  • ionizing radiation ionizing radiation.
  • Par-4 is involved in the apoptosis induced by BMP-9 in the PC-3 androgen insensitive prostate cancer cells (PC-3).
  • BMP-9 and BMP-10 are therapeutic agents in cancers including breast and prostate cancer.

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Abstract

L'invention concerne le rôle des protéines BMP-9 et/ou BMP-10 dans le traitement du cancer du sein ou de la prostate.
PCT/GB2007/002755 2006-07-29 2007-07-20 Activité anticancéreuse des protéines bmp-9 et bmp-10 et leur utilisation dans les traitements du cancer Ceased WO2008015383A2 (fr)

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WO2009084738A1 (fr) * 2007-12-28 2009-07-09 Kyowa Hakko Kirin Co., Ltd. Procédé et composition pharmaceutique pour le traitement du cancer utilisant la protéine bmp9
WO2009084739A1 (fr) * 2007-12-28 2009-07-09 Kyowa Hakko Kirin Co., Ltd. Procédé et composition pharmaceutique pour le traitement du cancer utilisant la protéine bmp10
WO2013152213A1 (fr) * 2012-04-06 2013-10-10 Acceleron Pharma, Inc. Méthodes et compositions pour l'augmentation des érythrocytes
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US9353356B2 (en) 2007-09-18 2016-05-31 Acceleron Pharma Inc. Activin-actriia antagonists for treating a follicle-stimulating horomone-secreting pituitary tumor
US9399669B2 (en) 2007-02-02 2016-07-26 Acceleron Pharma Inc. Variants derived from ActRIIB
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