ONCODEATH
Project no.: LSHC-CT-2006-037278
Project acronym: ONCODEATH
Project title: Sensitisation of solid tumour cells to death receptor
related therapies
Instrument: Sixth Framework Programme Specific Targeted Research
Project
Thematic Priority: FP6-2005-LifeSciHealth-6
Life Sciences, Genomics and Biotechnology for Health
Publishable Final Activity Report
Period covered: from 1/11/2006 to 30/4/2010 Date of preparation: 21 July 2010
Start date of project: 1st November 2006 Duration: 42 months
Project coordinator name: Alexander Pintzas
Project coordinator organisation name: National Hellenic Research Foundation
Executive summary
In the last decade, an encouraging decline of the death rate from cancer has been
observed, as a result of recent advances in prevention, early diagnosis and therapy.
Efforts are made towards the generation of “smart” anti-cancer drugs that will target
specific molecules depending on the molecular phenotyping of the patient’s tumour.
TNF-related apoptosis-inducing ligand (TRAIL) is currently a promising anti-tumour
agent with proven activity on several cell and animal cancer models. A unique feature of
TRAIL is its specificity towards malignant cells while sparing normal cells. At present,
preclinical studies with recombinant TRAIL are in progress as well as with an antiTRAIL-R2 monoclonal antibody with no hepatotoxicity. The aim of the present proposal
was to identify components in colon and other tumours which confer sensitivity or
resistance to cell death induced by TRAIL. The contribution of mutated oncogenes in
altering cell death molecules and pathways was examined on priority.
A large number of research tools were produced during : epitope-tagged expression
vectors, recombinant retroviruses-lentiviruses, antibodies. Cell lines with stably and/or
inducibly overexpressed or silenced by shRNA oncogenes as well of factors involved in
the signalling and apoptotic pathways in colon cancer have been generated. Quality
assessment of these tools in different applications had been completed. New transgenic
mouse models had been generated. New improved forms of TRAIL had been expressed
and utilised as well a panel of new small molecule kinase inhibitors had been synthesised
and selected for treatment. In addition, improved methods for cell apoptosis and in vivo
experiments had been developed and used for the analysis of apoptotic complexes as well
a for in vivo assessment of tumourigenicity. In addition, rational combinations of TRAIL
with inhibitors have been evaluated. Substantial progress has been made on deciphering
the interaction networks of oncogenic with apoptotic signalling pathways and trafficking
at the mitochondria level. On the mechanistic part, the network has identified
determinants for inhibitors in sensitizing to apoptosis as well as has provided detail
knowledge of the effect of the inhibitors on the intrinsic apoptotic amplification and
execution machinery related to caspases and Bcl-2 family members during TRAILinduced apoptosis. Assessment of sensitivity of tumours induced by activated oncogenes
in transgenic mice and nude mice to inhibitors alone or in the combination with the
recombinant mouse TRAIL has been examined.
The results of the network resulted in 12 published and 3 submitted joint publications as
well as 138 individual publications in high impact scientific journals like Cell, Cancer
Cell, Mol. Cell., Lancet Oncol., Science, EMBO J., Nature Cell Biol., Gastroenterology,
Cancer Res. Cell Death Differentiation, Oncogene, Br J Cancer and more. There are 8
new individual publications submitted-in preparation. The members of the network also
presented the major results in scientific conferences. Interaction between the network
participants was intense, which was realized via short-term visits, the annual meetings of
the network, bilateral meetings, via ftp site of the network and via electronic
communications. The project objectives have been advertised in the press like TV
channels, newspapers (please see section 2). The network’s web site is at:
http://www.eie.gr/nhrf/institutes/ibrb/eu-projects/oncodeath/index-en.html
1. Project execution
Section 1 – Project objectives and major achievements
Overall the network aimed
a. to contribute to better understanding of the role of specific activated oncogenes
or the expression pattern of certain proteins that can provide sensitivity or
resistance of tumour cells to TRAIL induced apoptosis
b. to identify rational combination of molecules inhibiting oncogene pathway
components with TRAIL agonists to achieve optimum synergistic effect.
The studies of the network were focused on : TRAIL-induced apoptosis, PI3K, TyK,
Aurora kinase and proteasome inhibitors, mitochondrial fission/fusion, caspase-2
The specific objectives were as follows:
1. Development of colon cell systems with stable expression of oncogenes known to
participate in the transformation of colonic epithelial cells.
-Preparation of colon cell lines with shRNA-induced down-regulation of transforming
oncogenes.
-Analysis of sensitivity of newly established colonic cell lines to TRAIL-induced
apoptosis.
2. Analysis of the status and activity of proximal components of TRAIL signalling
pathways
-Determination of a role of caspase-2 in TRAIL-induced apoptosis
-Characterization of oncogene-induced changes in mitochondrial events by TRAIL
3. Determination of the role of caspase-2 in sensitivity/resistance of tumour cells to
treatment.
-Analysis of determinants of Bax activation during TRAIL-induced apoptosis.
-Determination of a function of mitochondrial fission/fusion in TRAIL-triggered
apoptosis
4. Analysis of the effect of novel PI3K, TyK, Aurora kinase and proteasome inhibitors on
TRAIL-induced apoptosis
5. Evaluation of anti-tumour effects of selected inhibitors on mouse models of colon
carcinogenesis.
- Analysis of in vivo effects and synergism between selected inhibitors and TRAIL on
xenografts
-Selection of inhibitors that could cooperate with TRAIL in the induction of apoptosis
The network's objectives also included the development of new technologies and research
tools:
Generation of novel cell and animal systems
Battery of therapeutic factors:
a. the potent anti-tumour agent TRAIL and
b. combination of newly synthesised target based inhibitors of potent oncogenic pathways
like PI3K, Tyr kinase and Aurora kinase.
Generation of specific expression and silencing vectors (stable or inducible)
New imaging technology based on confocal microscopy to complement the sophisticated
biochemical analysis of pathways.
Fluorescence imaging in vivo technologies
The major achievements are as follows:
1. We have a collection of an essentially complete set of expression vectors and cell lines
overexpressing or lacking individual oncogenes.
3. We have prepared mutant forms of factors necessary to analyse interaction pathways
4. We have prepared different forms of tagged recombinant TRAIL
5. We have used newly synthesised PI3K, Hsp90, Aurora and choline kinase inhibitors
6. We have established a high-throughput method for apoptosis measurement for cell
lines using the INCell Analyzer
7. Direct orthotopic cell microinjection (OCMI) of human colorectal cancer cell lines in
animals has been worked out
8. We have generated triple transgenic mice allowing concomitant and inducible
expression of the K-RasV12G oncogene and deletion of both p53 alleles specifically in the
intestinal epithelium
9. We have isolated and characterised tumour initiating cells
10. Improved protocols of DISC complex detection have been worked out
11. Identification Arf and Rho GAP adapter protein ARAP1 as taking part in the
mobilization of TRAIL-R1/DR4 to the plasma membrane.
12. Discovery that TRAIL synergises with quercetin to induce cell death in colon cells
13. Discovery that DNA damage induces two distinct modes of cell death
14. Biological characterisation of newly synthesised PI3K, Hsp90, Aurora and choline
kinase inhibitors
15. Discovery that Aurora-A regulates nuclear factor-kappaB signaling by
phosphorylation of IkappaBalpha.
16. Discovery that genetic interference of choline kinase  by siRNA induces cell death
in human cancer cells but not in primary cells
17. Discovery that Bax activation and stress-induced apoptosis is delayed by the
accumulation of cholesterol in mitochondrial membranes.
18. Efficient protocols for treatment with TRAIL alone or in combination with novel
PI3K inhibitors
19. Identification of novel Aurora kinase inhibitor which synergises with TRAIL by
increased cleavage of Caspase 7 and PARP1
20. Killer TRAIL antitumour activity was demonstrated in colon cancer patient
xenograft/SCID mouse models in vivo.
21. Small cell lung carcinoma (SCLC) are resistant to TRAIL. Majority of human SCLC
cells are resistant to death receptor (DR)-mediated apoptosis because of defects in key
signaling molecules, such as DRs and/or initiator caspases. Combination of TRAIL and
anticancer agents might be beneficial for elimination of caspase-8-containing SCLC cells.
22. FLIP, AKT1 and AKT2 expression or activation were not the responsible of TRAILresistance in the colon lines analyzed.
23. A list of choline kinase inhibitors cooperating with TRAIL in inducing colon cancer
cell apoptosis has been generated.
24. Development and characterization of intestinal tumor generated in geneticallyengineered mice models
25. Establish an assay which evaluates live human tumor-derived cells for anticancer
drug response.
26. Role of caspase-2 as initiator caspase was also observed in apoptosis triggered by
other chemotherapeutic drugs. The present model for caspases-2 activation includes
PIDDosome complex formation.
27. Defining molecules/pathways of the intrinsic apoptotic machinery that are affected by
choline inhibitors and can potentially synergize with TRAIL in inducing apoptosis of
colon cancer cells.
28. Determination of the sensitivity of tumours implanted in immunosuppressed mice to
choline kinase inhibitors and assessment of their optimal dosing and in vivo synergism
with the recombinant human TRAIL.
Contractors involved
Partic
Role
*CO
Partic.
no.
1
*CR
2
CR
CR
3
4A
CR
4B
CR
CR
5
6
Participant name
Participant
short name
National
Hellenic NHRF
Research Foundation
Institute
Molecular IMG
Genetics,
Academy of
Sciences, Czech Republic
Karolinska Institutet
KI
Institute for
Cancer ICR/CCT
Research/Centre
for
Cancer
Therapeutics
Institute for
Cancer ICR/BCRC
Research/Breakthrough
Cancer Research Centre
University of Geneva
UniGe
Instituto
de IIB
Investigaciones
Biomédicas
Country
Greece
Czech
Republic
Sweden
U.K.
U.K.
Switzerland
Spain
CR
7
CR
8
Institut
CURIE - CNRS - Institut
UMR 144
Curie
GENEKOR A.E. Centre of GENEKOR
Research and Analysis of
Genetic Material (SME)
France
Greece
A detailed description of the work performed by each contractor during the reporting
period is included in the Periodic Management Report. This description is addressed at
the workpackage level.
NOTE:
Dr. George Nasioulas, the representative of the partner No 8 – YGEIA SA, has moved to
another company (GENEKOR) since 1/8/08 and he continued the project from his new
employment place. The relevant documents have been submitted and approved by EC.
Selected publications from the consortium
(ONCODEATH members are marked in bold)
Joint publications
Al-Saffar NM, Jackson LE, Raynaud FI, Clarke PA, Ramírez de Molina A, Lacal JC,
Workman P, Leach MO. The Phosphoinositide 3-Kinase Inhibitor PI-103
Downregulates Choline Kinase {alpha} Leading to Phosphocholine and Total Choline
Decrease Detected by Magnetic Resonance Spectroscopy. Cancer Res. 2010 Jul
1;70(13):5507-5517.
Oikonomou, E., Kosmidou, V., Katseli, A., Kothonidis, K., Mourtzoukou, D.,
Kontogeorgos, G., Andera, L., Zografos, G., and Pintzas, A. (2009). TRAIL Receptor
Upregulation Correlates to KRAS/ BRAF Mutations in Human Colon Cancer Tumours
and Respective Normal Tissue. Int. J. Cancer 125, 2127-2135
Psahoulia, F. H., Drosopoulos K. G., Doubravska, L., Andera, L. and Pintzas, A.
(2007). Quercetin enhances TRAIL-mediated apoptosis in colon cancer cells by inducing
the accumulation of death receptors in lipid rafts. Mol. Cancer. Ther 6, 2591-2599.
Oikonomou, E., Kothonidis, K., Taoufik, E., Probert, L. , Zografos, G., Nasioulas, G.,
Andera, L., and Pintzas, A. (2007). Newly Established Tumourigenic Primary Human
Colon Cancer Cell Lines are Sensitive to TRAIL Induced Apoptosis in vitro and in vivo.
Br. J. Cancer. 97, 73 – 84
Chan F, Sun C, Perumal M, Nguyen QD, Bavetsias V, McDonald E, Martins V, Wilsher
NE, Raynaud FI, Valenti M, Eccles S, Te Poele R, Workman P, Aboagye EO,
Linardopoulos S. Mechanism of action of the Aurora kinase inhibitor CCT129202 and
in vivo quantification of biological activity. Mol Cancer Ther. 2007, 6:3147-57
Joint manuscripts submitted for publication
Álvarez-Miranda M, Becerra M, Tébar LA, Ramírez de Molina A, Gómez del Pulgar T,
Andera L, Lacal JC. Combinatorial efficacy of TRAIL and MN58b against colon cancer
to overcome TRAIL resistance. Submitted.
Selected Individual publications
Montessuit, S., Somasekharan SP., Terrones O., Lucken-Ardjomande S, Herzig, S.,
Schwarzenbacher, R., Manstein, D., Bossy-Wetzel, E., Basanez, G., Meda, P., and
Martinou J.-C. (2010). Membrane Remodeling Induced by the Dynamin Related
Protein Drp1 Stimulates Bax Oligomerization. Cell. In press.
Vaculova, A., Kaminskyy, V., Jalalvand, E., Surova, O., and Zhivotovsky, B.
Doxorubicin and etoposide sensitize small cell lung carcinoma cells expressing caspase-8
to TRAIL. Mol Cancer. 2010: 91, 87.
Calzone, L., Tournier, L., Fourquet, S., Thieffry, D., Zhivotovsky, B., Barillot, E., and
Zinovyev, A. Mathematical modelling of cell-fate decision in response to death receptor
engagement. PLoS Comput. Biol. 2010: 6, e1000702.
Yap TA, Sandhu SK, Workman P, de Bono JS. Envisioning the future of early
anticancer drug development. Nat Rev Cancer. 2010 Jul;10(7):514-23.
Gaspar N, Sharp SY, Eccles SA, Gowan S, Popov S, Jones C, Pearson A, Vassal G,
Workman P. Mechanistic evaluation of the novel HSP90 inhibitor NVP-AUY922 in
adult and pediatric glioblastoma. Mol Cancer Ther. 2010 May;9(5):1219-33.
Workman P, Clarke PA, Raynaud FI, van Montfort RL. Drugging the PI3 kinome: from
chemical tools to drugs in the clinic.Cancer Res. 2010 Mar 15;70(6):2146-57. Review.
Zhivotovsky, B, and Orrenius, S. Clinical perspectives of cell death: where we are and
where to go…Apoptosis 2009: 14, 333-335.
Gogvadze, V., Orrenius, S. and Zhivotovsky, B. Mitochondria as targets for
chemotherapy. Apoptosis 2009: 14, 624-640.
Olsson, M., Vakifahmetoglu, H., Abruzzo, P.M., Högstrand, K., Grandien, A. and
Zhivotovsky, B. DISC-mediated activation of caspase-2 in DNA damage-induced
apoptosis. Oncogene, 2009: 28, 1949-1959.
Ott, M., Norberg, E., Zhivotovsky, B., and Orrenius, S. Mitochondrial targeting of
tBid/Bax: a role for the TOM complex? Cell Death Differ. 2009: 16, 1075-1082.
Gaspar N, Sharp SY, Pacey S, Jones C, Walton M, Vassal G, Eccles S, Pearson A,
Workman P. Acquired resistance to 17-allylamino-17-demethoxygeldanamycin (17AAG, tanespimycin) in glioblastoma cells. Cancer Res. 2009 Mar 1;69(5):1966-75.
Tondera D, Grandemange S, Jourdain A, Karbowski M, Mattenberger Y, Herzig S, Da
Cruz S, Clerc P, Raschke I, Merkwirth C, Ehses S, Krause F, Chan DC, Alexander C,
Bauer C, Youle R, Langer T, Martinou JC. SLP-2 is required for stress-induced
mitochondrial hyperfusion. EMBO J. 2009 Jun 3;28(11):1589-600.
Grandemange S, Herzig S, Martinou JC. Mitochondrial dynamics and cancer. Semin
Cancer Biol. 2009 Feb;19(1):50-6.
Hernando E, Sarmentero-Estrada J, Koppie T, Belda-Iniesta C, Ramírez de Molina V,
Cejas P, Ozu C, Le C, Sánchez JJ, González-Barón M, Koutcher J, Cordón-Cardó C,
Bochner BH, Lacal JC, Ramírez de Molina A. A critical role for choline kinase-alpha in
the aggressiveness of bladder carcinomas. Oncogene. 2009 Jul 2;28(26):2425-35.
Simova, S., Klima, M., Cermak, L., Sourkova, V., and Andera, L. (2008). Arf and Rho
GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the
plasma membrane. Apoptosis 13, 423-436.
Sayan, A.E., Sayan, B.S., Gogvadze, V., Dinsdale, D., Nyman, U., Hansen, T.M.,
Zhivotovsky, B., Cohen, G.M., Knight, R.A., and Melino, G. p73 and caspase-cleaved
p73 fragments localize to mitochondria and augment TRAIL-induced apoptosis.
Oncogene, 2008: 27, 4363-4372.
Eccles SA, Massey A, Raynaud FI, Sharp SY, Box G, Valenti M, Patterson L, de Haven
Brandon A, Gowan S, Boxall F, Aherne W, Rowlands M, Hayes A, Martins V, Urban F,
Boxall K, Prodromou C, Pearl L, James K, Matthews TP, Cheung KM, Kalusa A, Jones
K, McDonald E, Barril X, Brough PA, Cansfield JE, Dymock B, Drysdale MJ, Finch H,
Howes R, Hubbard RE, Surgenor A, Webb P, Wood M, Wright L, Workman P. NVPAUY922: a novel heat shock protein 90 inhibitor active against xenograft tumor growth,
angiogenesis, and metastasis. Cancer Res. 008 68:2850-60.
Lucken-Ardjomande S, Montessuit S, Martinou JC. Contributions to Bax insertion
and oligomerization of lipids of the mitochondrial outer membrane. Cell Death Differ.
2008 ;15 :929-37
Lucken-Ardjomande S, Montessuit S, Martinou JC. Bax activation and stress-induced
apoptosis delayed by the accumulation of cholesterol in mitochondrial membranes. Cell
Death Differ. 2008;15:484-93
Ramírez de Molina A, Sarmentero-Estrada J, Belda-Iniesta C, Taron M, Ramírez de
Molina V, Cejas P, Skrzypski M, Gallego-Ortega D, de Castro J, Casado E, GarciaCabezas MA, Sánchez JJ, Nistal M, Rosell R, González-Baron M,, Lacal JC..
Expression of choline kinase alfa to predict outcome in patients with early-stage nonsmall-cell lung cancer: a retrospective study. Lancet Oncol. 2007 Oct;8(10):889-97.
Raynaud FI, Eccles S, Clarke PA, et al., and Workman P. Pharmacologic
characterization of a potent inhibitor of class I phosphatidylinositide 3-kinases. Cancer
Res., 2007, 67:5840-5850
Sharp SY, Boxall K, Rowlands M, Prodromou C, Roe SM, Maloney A, Powers M,
Clarke PA, et al., and Workman P. In vitro biological characterization of a novel,
synthetic diaryl pyrazole resorcinol class of heat shock protein 90 inhibitors. Cancer Res.,
2007, 67 :2206-2216
Sun C, Chan F, Briassouli P, Linardopoulos S.Aurora kinase inhibition downregulates
NF-kappaB and sensitises tumour cells to chemotherapeutic agents. Biochem Biophys Res
Commun., 2007;352:220-225.
Janssen K-P., P. Alberici, H. Fsihi, C. Gaspar, C. Breukel, P. Franken, C. Rosty, M. Abal,
F. El Marjou, R. Smits, D. Louvard, R. Fodde, and S. Robine. The APC and oncogenic
KRAS are synergistic in enhancing Wnt signaling in intestinal tumor formation and
progression. Gastroenterology, 2006, 131:1096-1109.
Fevr T, Robine S, Louvard D, Huelsken J.Wnt/beta-catenin is essential for intestinal
homeostasis and maintenance of intestinal stem cells. Mol Cell Biol., 2007, 21:7551-9.
Vignjevic D, Schoumacher M, Gavert N, Janssen KP, Jih G, Laé M, Louvard D, BenZe'ev A, Robine S. Fascin, a novel target of beta-catenin-TCF signaling, is expressed at
the invasive front of human colon cancer. Cancer Research, 2007, 67:6844-53.
Cattaneo F, Molatore S, Mihalatos M, Apessos A, Venesio T, Bione S, Grignani P,
Nasioulas G, Ranzani GN. Heterogeneous molecular mechanisms underlie attenuated
familial adenomatous polyposis. Genet Med. 2007;9 :836-841
Simova, S., Klima, M., Cermak, L., Sourkova, V., and Andera, L. (2008). Arf and Rho
GAP adapter protein ARAP1 participates in the mobilization of TRAIL-R1/DR4 to the
plasma membrane. Apoptosis 13, 423-436.
Tamm, C., Zhivotovsky, B., and Ceccatelli, S. Caspase-2 activation in neural stem cells
undergoing oxidative stress-induced apoptosis. Apoptosis, 2008: 13, 354-363.
Gogvadze, V., Orrenius, S., and Zhivotovsky, B. Mitochondria in cancer cells: what is
so special about them? Trends Cell. Biol., 2008: 18, 165-173.
Sayan, A.E., Sayan, B.S., Gogvadze, V., Dinsdale, D., Nyman, U., Hansen, T.M.,
Zhivotovsky, B., Cohen, G.M., Knight, R.A., and Melino, G. p73 and caspase-cleaved
p73 fragments localize to mitochondria and augment TRAIL-induced apoptosis.
Oncogene, 2008: 27, 4363-4372.
Vaculova, A., and Zhivotovsky, B. Caspases: determination of their activities in
apoptotic cells. Meth. Enzymol. 2008, 442, 157-181.
Shen, J., Vakifahmetoglu, H., Stridh, H., Zhivotovsky, B., and Wiman, K.G. PRIMA1MET induces mitochondrial apoptosis through activation of caspase-2. Oncogene, 2008:
27, 6571-6580.
Norberg, E., Gogvadze, V., Ott, M., Horn, M., Uhlén, P., Orrenius, S., and Zhivotovsky,
B. An increase in intracellular Ca2+ is required for the activation of mitochondrial
calpain to release AIF during cell death. Cell Death Differ. 2008: 15, 1857-1864.
Powers MV, Clarke PA, Workman P. Dual targeting of HSC70 and HSP72 inhibits
HSP90 function and induces tumor-specific apoptosis. Cancer Cell. 2008, 14 :250-62
Vaughan CK, Mollapour M, Smith JR, Truman A, Hu B, Good VM, Panaretou B,
Neckers L, Clarke PA, Workman P, Piper PW, Prodromou C, Pearl LH. Hsp90dependent activation of protein kinases is regulated by chaperone-targeted
dephosphorylation of Cdc37. Mol Cell. 2008, 31:886-95
Eccles SA, Massey A, Raynaud FI, Sharp SY, Box G, Valenti M, Patterson L, de Haven
Brandon A, Gowan S, Boxall F, Aherne W, Rowlands M, Hayes A, Martins V, Urban F,
Boxall K, Prodromou C, Pearl L, James K, Matthews TP, Cheung KM, Kalusa A, Jones
K, McDonald E, Barril X, Brough PA, Cansfield JE, Dymock B, Drysdale MJ, Finch H,
Howes R, Hubbard RE, Surgenor A, Webb P, Wood M, Wright L, Workman P. NVPAUY922: a novel heat shock protein 90 inhibitor active against xenograft tumor growth,
angiogenesis, and metastasis. Cancer Res. 008 68:2850-60.
Consortium management
The program has its own internet site, containing all important information, which is
updated with useful information regarding project progress. The address is:
http://www.eie.gr/nhrf/institutes/ibrb/eu-projects/oncodeath/index-en.html
The main forums of scientific exchange and organization are the Annual Meetings.
The fourth Annual Meeting was organised in Madrid, Spain in September 11-12, 2009
and the final Meeting was organised in Geneva, Switzerland in March 26-27, 2010
2. Dissemination and use
Publishable results of the Final plan for using and disseminating the knowledge (see
format in Appendix 1).
Section 1 - Exploitable knowledge and its Use
Final plan for using and disseminating the knowledge
Until recently, the approaches to identifying new cancer treatments have been relatively
unsophisticated, relying on non-specific strategies that could kill both tumour cells and
normal cells. New anticancer drugs would be identified by measuring the effect on cancer
cells growth or inhibition in experimental animals. However, a convergence of scientific
advances such as bioinformatics, pharmacogenomics and molecular cancer medicine has
facilitated the identification of unique molecular targets specific to cancer cells. With an
increased understanding of the biological basis for cancer onset and progression, the
molecular changes that distinguish malignant cells from normal cells are becoming
increasingly apparent, offering a growing range of potential drug targets in the form of
altered genes, proteins or corrupted pathways. The increased selectivity offered by these
unique targets offers developers the opportunity to cultivate more efficacious and less
toxic “molecular-targeted” treatments. These new opportunities have not gone un-noticed
by the big Pharma corporations, resulting in extensive R&D activities and the very
successful marketing of archetypal drugs such as Gleevec (developed and marketed by
Novartis with global sales of $3 billion in 2007), Herceptin, Avastin and Rituxan
(developed and marketed by Genentech and Roche with annual sales of $4 billion, $3.4
billion and $2.28 billion respectively in 2007). GSK (Tykerb) and Amgen (Vectibix) also
launched targeted cancer drugs towards the end of 2006 and the beginning of 2007 with
sales for 2007 already reaching $100 million and $170 million respectively.
Already many publications document sensitizing effects of a number of drugs and
compounds as DNA damaging agents, proteasome or histone deacetylases inhibitors and
dozens of various kinase inhibitors on TRAIL-induced apoptosis, but only fraction of
them will be eventually used for a TRAIL-based, complex cancer therapies. Our finding
of two more compounds Manumycin A and Homoharringtonine that can sensitize cancer
cells to TRAIL-induced apoptosis might just only add them to numerous other
“sensitizers” that would never make it to clinical use or they could be also few of those
potentially used for cancer therapy. TRAIL can also synergise with quercetin to
overcome resistance in colon cancer cells.
We conclude that, chemical inhibition of Aurora kinases does not significantly sensitise
cells to TRAIL when the inhibitors are used at low concentrations. However, at higher
concentrations (
apparent change in mitotic spindle morphology. Similarly, drugs that interfere with
tubulin function sensitise cells to TRAIL at the concentrations necessary to effectively
interfere with spindle morphology. Based on this data, we propose that albendazole and
fenbendazole, which are currently in clinical trials as alternatives to taxol, can also be
used as sensitisers to TRAIL in the clinic.
Contribution of ONCODEATH towards personalised cancer therapies:
Conventional cancer treatments have been based mostly on the "one size fits all"
approach. Patients are prescribed medicine in standard doses that are later adjusted by a
"trial and error" process. Patients may suffer severe side effects, and while some respond
well to the treatment, others show no improvement. Innovations in the field of genetics
have led to a new, personalized treatment approach where understanding the genetic basis
of the individual patient and his tumor help determine the patient's eligibility to a
proposed treatment and predict its success. The advantages of this approach to patients
are self-evident. ONCODEATH has used the approach of identifying two cancer drugs
under development such as TRAIL and choline kinase, PI3K and BRAF inhibitors as a
novel combinatorial strategy. Our findings where a simultaneous combinatorial treatment
of choline kinase inhibitors and TRAIL shows a strong synergistic effect can provide an
alternative treatment to those used in the clinic today.
Sensitisation findings have been observed by combinatorial treatment of TRAIL with
BRAF or hsp90 inhibitors, depending on the tumour mutation profile. TRAIL and
TRAIL-related reagents (currently agonistic anti-TRAIL receptors antibodies, and
potentially also small molecules/peptides substituting in activating the apoptotic
signaling) are already being recognized and tested as an important and likely very
effective, additional weapon in our “never-ending” war against malignant
transformations. As with all weapons/tools, their effective and safe use is linked with indetail understanding of their function/signaling (i.e. characterization and regulation of
TRAIL-induced signaling in cancer cells) as well as provided support (drugs or
compounds) that can safely enhance TRAIL-induced elimination of tumor cells in cancer
patients. Thus, our characterization of “mode of action” of selected colorectal cancerrelated oncogenes in the regard to TRAIL-induced apoptosis (see below) could be used
for better understanding and implementation of future TRAIL-related treatment strategies
of cancer patients:
BRAF inhibitors cooperate with TRAIL to overcome PI3K resistance in colorectal cancer
cells. Data suggest that not only BRAF but also PIK3CA when found as single driver
mutations may confer sensitivity to TRAIL. Successful pharmacological application of
either BRAF inhibitor as single agent was limited. Co- Treatment of BRAF inhibitors
with PI3K inhibitors and/or TRAIL can overcome PI3K resistance, depending on the
mutation status of the tumour.
The decreased expression or activation of survival key molecules followed by PI-103 or
17-AAG treatments together with TRAIL strongly contributed to apoptosis in colorectal
cancer cells providing not only a biological mechanistic explanation but also an
indication for bio-markers to be possibly used in future clinical trials.
We can conclude that inducing apoptosis by TRAIL in combination with HSP90 or PI3K
inhibitors may represent a potent therapy in colorectal cancer as an alternative to current
treatments both for its efficacy and potential absence of toxicity.
The WHOs “World Cancer Report” anticipates a greater than 50% increase in the annual
global incidence of cancer, from 10 million to 15 million cases, over the first three
decades of the 21st century. Additionally incidents of deaths from cancer in the world are
projected to continue rising, from 7.6 million in 2005 to an estimated 9 million people
dying from cancer in 2015 and 11.4 million in 2030. As a consequence, cancer will
continue to take on an increasingly prominent role in shaping healthcare policy around
the globe. The social, economic and political repercussions of this continued rise in
disease incidence will be profound.
On the other side, in the last ten years, the average annual growth in total cancer-related
health expenditure has been 7.9%. The global oncology drug market spending was more
than $28 billion in 2005 and grew by 18.6% in 2005 compared with 2004. The fact that
the market has seen such growth while the patient base remained steady shows that there
is a great thirst for a new treatment that will make more than merely incremental
improvements to the patient’s condition, quality of life and life expectancy.
The drug industry is dominated by giants, such as Johnson & Johnson, Lilly, Merck, GSK
and Pfizer, as they alone have the resources to conduct drug development. Tufts
University's Center for the Study of Drug Development estimates the expected cost of
developing a single new drug be $802 million. However, new data shows that the
development cost of a new drug can vary from around $500 million to $1.2 billion.
In terms of sales, oncology is currently the third largest therapy area franchise, behind the
cardiovascular (CV) and central nervous system (CNS) markets. The sector is currently
experiencing strong growth driven by increased diagnoses of cancer, continuing unmet
needs within the marketplace, and the escalating costs of treatment. According to the IMS
Global Oncology Forecast, sales of oncology products in 2008 exceeded $48 billion,
contributing nearly 17 percent of global pharmaceutical sales growth for this year. Sales
of cancer drugs are expected to continue growing at nearly double the rate of the global
pharmaceutical market and reach $80 billion by 2011. According to IMS, development of
new treatments, an increasing number of patients on chemotherapy in major markets and
evidence that more people in emerging markets are gaining access to modern targeted
therapies will contribute to sales of cancer drugs growing at a compound rate of 12 to 15
percent. Moreover, the market segment of kinase inhibitor drugs of the type developed by
TCD, is growing fast and seems to be in favor with the multinational pharmaceutical
companies. According to a report by the Business Communications Company sales of
kinase inhibitors researched $4.6 billion in 2005 but sales are expected to exceed 12.6
billion in 2010 with an annual growth of 20.6%.
Thus, the potential market for new rational combinatorial strategies of combining
TRAIL. choline kinase , PI3K, BRAF kinase inhibitors is huge and may have a
tremendous economical impact.
Section 1 - Exploitable knowledge and its Use
This section will only present exploitable results, defined as knowledge having a
potential for industrial or commercial application in research activities or for
developing, creating or marketing a product or process or for creating or
providing a service.
It should provide an overview, per exploitable result, of how the knowledge
could be exploited or used in further research (if relevant). This should be created
by the project coordinator obtaining input from each contractor that owns the
knowledge and has an active role in its exploitation.
Both past and planned future activities should be included.
Where applicable please also include an explanation of why planned activities
mentioned in previous reports have been discontinued or altered.
Exploitable
Knowledge
(description)
1. New
sensitizers of
colon cancer
cells to TRAILinduced
apoptosis.
Exploitable
product(s) or
measure(s)
Mode of action
of
homoharringto
nine and
manumycin A
in sensitizing to
TRAIL-induced
apoptosis.
Sector(s) of
application
1. Medical
Timetable for
commercial
use
2011-2012
Owner & Other
Partner(s)
involved
Patents or other
IPR protection
Application for Partic. IMG
the patent
(owner)
concerning new
sensitizer of
TRAIL-induced
apoptosis is
planned for
2011.
We have identified two new sensitizers of TRAIL-induced apoptosis of TRAILresistant colorectal carcinoma cells. This finding is potentially extendable to
TRAIL-resistant cells from tumors of other than colorectal origin. The efficacy of
these new TRAIL sensitizers needs to be evaluated in animal models and
eventually in pre-clinical tests. The characterization of these two compounds has
been done mainly at IMG.
Section 2 – Dissemination of knowledge
The dissemination activities section should include past and future activities and
will normally be in the form of a table maintained by the coordinator or any other
person charged with controlling the dissemination activities.
Overview table
Planned/ac
tual
Dates
Type
Jan 2007
Press release (TV)
April 2010
Press release
(newspaper)
Media briefing
1.3.10.2009
Conferenc
organisation- MAC’09
During
period
Conference
participation (65- for
details, see final
management report))
Exhibition
Publications (153 - for
details, see final
management report)
Project web-site
http://www.eie.gr/nhrf/i
nstitutes/ibrb/euprojects/oncodeath/ind
ex-en.html
Posters (45 in total-for
details , please see final
management report)
Type of
audience
General
public
General
public
Higher
education
Researchers
and PhD
students
Research
Countries
addressed
Size of
audience
Greece
National
TV chann
Nationwide
Partner
responsi
ble
/involved
NHRF
(CO)
NHRF
(CO)
160
IMG
Greece
European
and oversees
counrties
Several
All
partners
Industry (
sector x)
All
partners
NHRF
(CO)
All
partners
IMG has been a co-organizer of an international meeting Mitochondria, apoptosis
and Cancer 2009, which took a place in Prague, Oct 1-3, 2009 with the
participation of 160 researchers from both European and oversees countries.
Results from the Oncodeath project (New sensitizers to TRAIL-induced
apoptosis) were presented in a form of poster.