STAND-ALONE PROJECT - FINAL REPORT

STAND-ALONE PROJECT - FINAL REPORT Project number P 21630-N22 Project title Weiterführende Erforschung der Strahlenwirkung kleiner Dosen Project leader PD Dr. Helmut Schöllnberger Project website 1 http://www2.sbg.ac.at/fwf2/ 1 Projects that started after 01.01.2009 are required to have a website Final Report February 2009/ Page 1 of 15

1. Summary for public relations work The main finding of FWF project P 21630-N22 of Priv.-Doz. Dr. Helmut Schöllnberger is that the carcinogenic process that leads to the formation of the first cancer cell in lung and colon tumors is dominated by the growth of mutated cells, whereas the mutation rates have less effect. This significant finding that has importance for cancer research was achieved in an international collaboration with top experts from Imperial College London, ETH Zürich, and RIVM in the Netherlands. The FWF Project of Dr. Schöllnberger, until September 2009 working at the University of Salzburg, was mainly concerned with questions related to the origin of the first cancer cell in lung and colon cancer. In a simplified way, the formation of a cancer cell can be imagined as follows: out of a pool of normal stem cells one cell receives a critical mutation leading to a mutant cell. The latter has a selective growth advantage compared to the normal cells leading to a clone of mutated cells. If one of these mutated cells receives another critical mutation, then this cell converts into a malignant cell that may subsequently grow into a clinically detectable tumor. Since decades, it is a much disputed question in cancer research what the most important process is in the formation of the first malignant cell, mutation induction or the clonal growth of mutated cells. Two different state-of-the-art mathematical cancer models were used to investigate this question. One model is an evolutionary concept of tumorigenesis, the other one is the well-know stochastic two-stage model. While both models are very different from a mathematical point of view, they share important features. Both explain the appearance of the first malignant cancer cell by accumulating mutations with a selective advantage in a cell population, and both distinguish the generation of a new (mutated) cell type from its growth (i.e. the clonal expansion of mutated cells). The dynamics of the carcinogenic process can be directly compared between the two models in terms of the time to appearance of the first malignant cell. For each of the two models a mathematical formula was developed that gives the time until 50% of a population of individuals harbour a first cancer cell which can grow into a malignant cancer. In both formulae the parameters governing the growth of clones of mutated cells that contain advantageous mutations enter the equation directly, whereas the mutation rates have a much smaller influence. Thus, in both models, the selection parameters have the strongest impact on the waiting time equations, suggesting that cell selection is the major driving force of carcinogenesis. The observed prominent role of cell selection was confirmed by numerical evaluation of the waiting time equations using best estimates from various model fits to epidemiological data and suitable values from the scientific literature. Dr. Schöllnberger and his co-investigators expect the main finding to hold also for other solid tumors. The study has been published in 2010 in Cancer Research, one of the most prestigious scientific journals in this field. The study relates to the biological mechanisms of tumor cell formation and therefore represents basic research. Possible implications for medical areas such as patient treatment are unclear. Final Report February 2009/ Page 2 of 15

Zusammenfassung: Das FWF Projekt P 21630-N22 von Priv.-Doz. Dr. Helmut Schöllnberger hatte als wichtigstes Ergebnis die Erkenntnis, dass der karzinogene Prozess der Bildung einer ersten Krebszelle eines Lungenkarzinoms oder eines Darmkrebses dominiert wird durch das Wachstum von mutierten Zellen, während die Mutationen selbst einen viel kleineren Einfluss haben. Dieses für die Krebsforschung bedeutende Ergebnis wurde im Rahmen einer internationalen Kollaboration mit Wissenschaftlern vom Imperial College London, der ETH Zürich und des RIVM, Niederlande, erzielt. Das Projekt von Dr. Schöllnberger, bis September 2009 an der Universität Salzburg tätig, war hauptsächlich mit Fragen beschäftigt, die sich auf die Entstehung der ersten Krebszelle eines Lungenkarzinoms oder Darmkrebses beziehen und stellt daher Grundlagenforschung dar. Vereinfacht kann man sich die Krebsentstehung so vorstellen, dass in einer großen Anzahl von gesunden Stammzellen eine Zelle eine kritische Mutation erhält. Dies führt zu einer mutierten Zelle, die einen Wachstumsvorteil gegenüber den normalen Stammzellen hat. Auf diese Weise entsteht ein Klon von mutierten Zellen. Wenn eine dieser mutierten Zellen eine weitere kritische Mutation erfährt, dann wird aus dieser Zelle eine maligne Zelle aus der in weiterer Folge ein klinisch feststellbarer Tumor entstehen kann. Seit Jahrzehnten wird in der Krebsforschung die Frage diskutiert, welcher Prozess bei der Entstehung einer Krebszelle der wichtigste ist, die Induktion von Mutationen oder das Wachstum von mutierten Zellen. Zwei hochstehende mathematische Krebsmodelle wurden verwendet, um diese Frage zu klären. Eines ist ein evolutionäres Krebsmodell, das andere ist das bekannte stochastische Zweistufenmodell. Während beide Modelle mathematisch sehr unterschiedlich sind, haben sie aber auch essentielle konzeptionelle Ähnlichkeiten. Beide erklären das Auftreten der ersten Krebszelle durch die Akkumulation von Mutationen innerhalb einer Zellpopulation. Diese Mutationen stellen einen Selektionsvorteil dar. Beide Modelle unterscheiden die Erzeugung eines neuen (mutierten) Zelltyps vom Wachstum der mutierten Zellen. Die Dynamik des karzinogenen Prozesses kann in beiden Modellen durch die Zeit bis zur ersten Krebszelle dargestellt werden. In der Studie wurde für jedes Modell eine Formel berechnet, die angibt wie lange es dauert bis die Hälfte einer großen Anzahl von Personen eine erste Krebszelle in sich trägt. In beiden mathematischen Gleichungen gehen jene Modellparameter, die das Wachstum der mutierten Zellklone beschreiben, direkt ein, während die Mutationsraten einen viel schwächeren Einfluss haben. In beiden Modellen haben also die Selektionsparameter den stärksten Einfluss auf die sog. Wartezeit bis zum Tumor, d.h. dass die Zellselektion die wichtigste treibende Kraft ist bei der Entstehung von Krebszellen. Untermauert wird dieses Ergebnis durch die Evaluierung der beiden Formeln mit numerischen Ergebnissen aus Anpassungen des Zweistufenmodells an epidemiologische Daten und geeigneter Werte aus der Literatur. Dr. Schöllnberger und seine Koauthoren erwarten, dass das vorliegende Ergebnis nicht nur für Lungen- und Darmkrebs Gültigkeit hat, sondern auch für andere solide Tumore. Die Studie wurde 2010 in der Zeitschrift Cancer Research publiziert, einer der weltweit führenden wissenschaftlichen Zeitschriften für Krebsforschung. Mögliche Auswirkungen auf die Medizin wie zum Beispiel die Behandlung von Tumorpatienten sind unklar. Final Report February 2009/ Page 3 of 15

2. Brief project report 2.1 Report on the scientific work 2.1.1 information on the development of the research work It is pointed out that the entire FWF project effectively lasted only for 3 months: 1.7.2009 30.9.2009. Because of mayor funding problems between the end of his first FWF project and the beginning of the second one, Dr. Schöllnberger on Oct. 1 2009 took a new position at Helmholtz Zentrum München (HMGU). Therefore, he therefore interrupted the project for 12 months. After that he terminated the FWF project to continue working on a different project at HMGU. The original FWF grant proposal focused mostly on the development of a new model for in vitro neoplastic transformation and testing that model on the data by Redpath et al. (Rad Res 156: 700-707, 2001). In 2006, however, Dr. Schöllnberger was invited by Dr. Paolo Vineis, Imperial College, to participate in a fascinating study that should compare two state-of-the-art cancer models to find out which process in the formation of the first cancer cell of a solid tumor is the most important one. Dr. Schöllnberger took up this opportunity app. in April 2006 because it offered manifold opportunities to scientifically interact with two other colleagues, mathematicians at ETH Zürich and RIVM, on sophisticated mathematical aspects related to these cancer models. These colleagues were Dr. Beerenwinkel and R.Hoogenveen. During the three months in 2009 this study was finalized and a manuscript written and submitted for publication. The scientific goal of the study was to investigate whether promotion (i.e. the clonal expansion of mutated cells) could be the most important biological process in the carcinogenic formation of the first cancer cell. This was implicated by numerical findings in a study of Beerenwinkel et al. (PLoS Comput Biol. 3(11): e225, 2007). During the 3 months duration of the project, a manuscript has been written and submitted to PNAS. Parallel to the writing of the manuscript final calculations related to the study were performed. The manuscript was finally published in Cancer Research: Schöllnberger et al. Cancer Research 70: 6797-6803, 2010. Progress has also been made on the work related to the new model for in vitro neoplastic transformation which had earlier been sketched with scientific advice from Dr. Robert Stewart. The model has been equipped with low-dose hyper-radiosensitivity and bystanderinduced apoptosis and includes radiation-induced and endogenously produced simple and complex lesions. Preliminary fits to the in vitro data from Redpath et al. (2001) have been performed. Because of the new position at HMGU, Dr. Schöllnberger has not been able to finalise and publish that study so far. 2.1.2 most important results and brief description of their significance (main points) with regard to contribution to the progress of the field (e.g. did the results contribute to an increased importance for the field and in what way?); breaking of new scientific ground (i.e. to what extent and in what respect?); most important development of hypotheses (what relevance did the project have for the development of scientific hypotheses, e.g. were new hypotheses developed or old hypotheses disproved?); development of new methods; relevance for other (related) areas of science (transdisciplinary issues and methods). Final Report February 2009/ Page 4 of 15

It has been a long disputed question in cancer research what the most important process is in the formation of the first cancer cell of a solid tumour (see e.g. Tomlinson IP, Novelli MR, Bodmer WF. The mutation rate and cancer. Proc Natl Acad Sci USA. 93(25): 14800-3, 1996). The project yielded the groundbreaking finding that the most important biological process in the formation of the first cancer cell (that will subsequently grow into a clinically detectable tumour) is the growth of mutated cells (i.e. the selective advantage) and not the induction of mutations. This was achieved by calculating the median time to cancer for the so called S-MVK model (simplified Moolgavkar-Venzon-Knudson model), S-MVK. The resulting formula, which gives the time until 50% of a population of individuals contain a first cancer cell (starting the process with N normal stem cells), clearly shows that the rate of net clonal expansion of initiated cells ( - ) has a much stronger influence than the mutation rates 1 and 2 : S MVK 2 1 ( ) ln2 ln. ( ) 1 2N (1) This result is strikingly similar to another expression for the median time to cancer derived by Schöllnberger et al. (2010) for the Wright-Fisher process, an evolutionary stochastic cancer model: 1/ k 2 k!ln 2 ln [ s /( ud)] WF. (2) N 2sln N clone Like for the S-MVK model, the process is started with N normal cells. Here, s is the selective advantage per mutation. It enters the equation directly while the per-gene mutation rate u enters the equation logarithmically. The number of putative cancer associated genes involved in the process is denoted by d. A cell is considered initiated if k = 5 of those genes are mutated and malignant if k = 20. N clone is the mean number of adenoma cells (the latter correspond to initiated cells in the MVK model). In addition to these equations, the median time to cancer starting with a clone of m initiated (mutated) cells has been calculated using the stochastic MVK model. The result is similar to Eq. (1), i.e. the rate of net clonal expansion ( - ) has a much stronger influence than the mutation rate 2. These equations and the mechanistic insight that they allow are the main finding of the project. In addition, Dr. Schöllnberger and his colleagues analyzed whether the predicted time from a clone of m initiated cells to the first malignant cell is consistent between the two models. For Eq. (1) these numerical evaluations were performed using various sets of best estimated values (obtained from other studies) from fits of the MVK model to epidemiological data for lung and colon cancer. Eq. (2) was evaluated using biologically plausible values for k, d, u, and s from the paper by Beerenwinkel et al. (2007): the selective advantage per mutation is on the order of 1 10 3 to 1 10 4 ; the average per-gene mutation rate has been estimated as u = 1 10-7 per generation; and d = 100. When the process is started with one mutated cell (i.e. m = 1), the following values were obtained using the MVK model: between 99 and 192 years for lung cancer, and between 62 and 69 years for colon cancer. For the Wright-Fisher process, the resulting waiting times are between 18 and 44 years. The approximations made in the derivation of Eq. 2, together with the fact that it had to be evaluated with parameter values that were not determined by fitting cancer incidence or mortality data, make the comparison with the MVK model difficult. It is therefore not surprising that somewhat different numerical values are obtained. However, all results are Final Report February 2009/ Page 5 of 15

clearly in the same order of magnitude, and given the approximations that were necessary to derive Eq. (2), they appear in reasonable agreement. Summarizing it can be said that for both models, the mathematical expressions for the median waiting time to the first malignant cell clearly show the dominating impact of cell selection (i.e., cell proliferation) on carcinogenesis. The mutation rates enter the equations for the median time to cancer only logarithmically while the selection parameters enter the equation directly. Because the finding stems from the use of two (very) different mathematical concepts of carcinogenesis, the discovery is especially valuable. Dr. Schöllnberger and his co-investigators are confident that the study adds an important step towards a final answer to the question what the most important driving force is in carcinogenesis. An old hypothesis in cancer research is that mutation induction is the most important process in the formation of the first malignant cell. The finding reported here is an important step towards disproving this old hypothesis and is in line with results from previous studies (e.g. Tomlinson et al. Proc Natl Acad Sci USA. 93: 14800-14803, 1996). To the best of our knowledge it is the first time that two state-of-the-art stochastic cancer models were mathematically compared to each other which led to a fascinating new insight into the formation of the first cancer cell of solid tumours. The relevance for other (related) areas of science is unknown at the moment. 2.1.3 information on the running of the project, use of the available funding and where appropriate any changes to the original project plan relating to (2000 characters excl. spaces) Duration: The project lasted for 3 months (July 1 2009 Sept. 30 2009). Between the end of his first FWF project (P18055-N02) on Dec 31 2008 and the beginning of the 2 nd FWF project on July 1 2009, the project leader had mayor funding problems because the follow-up FWF grant proposal was rejected twice (despite very thorough revisions that were perform). Dr. Schöllnberger tried to get financial support from the officials at Salzburg University and from the government of Salzburg. Such support was not granted. Because of this problematic situation, Dr. Schöllnberger took a new position at HMGU and started working on a different project. It is delicate to note that in a country like Austria, which is surrounded by several nuclear power plants and in which a majority of its population is afraid of radiation (especially since the Chernobyl disaster), it was not possible to get limited financial support for the only Austrian scientist who has been working in the field of cancer risk estimation related to low and medium doses since the beginning of his dissertation in 1994 until his Habilitation in 2008 and beyond. Use of personnel: Larger items of equipment purchased; Final Report February 2009/ Page 6 of 15

Other significant deviation 2. 2.2. Personnel development importance of the project for the scientific careers of those involved (including the project leader) Brief comments on particular developments: The project led to a publication in Cancer Research (including extensive Supplementary Material published online) which is one of the leading scientific journals in this field. It currently has an impact factor of 7.543 and is therefore Dr. Schöllnberger s most prestigious publication so far. http://cancerres.aacrjournals.org/site/misc/infoforlibrarians.xhtml Special possibilities opened up by the project (e.g. high-risk topics, intensification of transdisciplinary research etc.): A follow-up study was opened up by the project: for the publication in Cancer Research the median time to failure was derived for the S-MVK model (Eq. (1): S-MVK ), a deterministic variant of the stochastic MVK model. In addition, the study led to an analytical solution for the median time to failure for the stochastic MVK model for = 0 (i.e. no extinction of clones). Personal communication with Dr. E. Georg Luebeck, Fred Hutchinson Cancer Research Centre, convinced the project leader that it would be useful to have an analytical solution for the median time to failure for the MVK model for 0, i.e. MVK. Currently, Dr. Schöllnberger derives this analytical solution together with Rudolf Hoogenveen from RIVM. The solution will be evaluated using the results of model fits, which have also been used by Schöllnberger et al. (Cancer Research 2001). That will lead to interesting comparisons between the values for S-MVK and MVK. Importance (effect, success) of these new possibilities: This new research study is an exciting opportunity for the project leader and for R. Hoogenveen to publish a second paper in Cancer Research. That would not only add to their scientific reputation but also increase the scientific impact of their first Cancer Research paper. Effects of the project and of research performed in the framework of the project on the international reputation of the project leader and co-workers. The publication in Cancer Research certainly helped to improve Dr. Schöllnberger s scientific reputation and that of his co-investigators. Establishment or intensification of international collaborations: Earlier collaborations with Dr. Vineis and Rudolf Hoogenveen have been intensified through the project. A new fruitful collaboration with Dr. Niko Beerenwinkel was established. 2 The assessment of what should be regarded as a significant deviation is the responsibility of the project leader. As a guideline, any deviation of more than 25% from the original financial plan or work schedule should be accounted for. Final Report February 2009/ Page 7 of 15

2.3 effects of the project outside the scientific field (2000 characters excl. spaces) Brief comments on particular developments (perspectives); Organization of symposiums and conferences; Did the project have any particular relevance for developments in teaching and if so, what (incorporation of project results in lectures, seminars etc.)? Particular public relations activities for the general public 1.) The summary of this report will be released to the media in Austria via the Austria Press Agency (APA). 2.) A new webpage at has been installed to help disseminate the scientific findings: http://www2.sbg.ac.at/fwf2/index.html This webpage is linked to the project leader s webpage at Salzburg University: http://www.uni-salzburg.at/portal/page?_pageid=805,269875&_dad=portal&_schema=portal The Cancer Research publication and related Supporting Information have also been posted at http://www.uni-salzburg.at/portal/page?_pageid=805,269875&_dad=portal&_schema=portal (to find this webpage search Google for Schoellnberger ). 3.) In the next few days, the German summary of this final report will be sent to the public relations office at the University of Salzburg to be published as follows: a) At the University s homepage: http://www.uni-salzburg.at b) In the electronic newsletter of Salzburg University c) As a contribution to the Salzburger UNI Nachrichten which will be published as an attachment to the Salzburger Nachrichten, the local newspaper in the province of Salzburg. The homepage and electronic newsletter reach app. 2700 employes at the University of Salzburg and app. 16500 students. The Salzburger UNI Nachrichten reach app. 250000 readers of Salzburger Nachrichten in Salzburg/Austria. d) The public relations office of Salzburg University will also send out a press release related to the publication in Cancer Research. Based on that, local median such as Salzburger Nachrichten and ORF, the Austrian television and radio network, can decide whether they would like to publish an article about Dr. Schöllnberger s study. 5.) In the next few days, the German abstract of this final report will also be sent to the public relations office of Helmholtz Zentrum München to be published at the HMGU homepage: http://www.helmholtz-muenchen.de/ and in HMGU Newsletter http://nip.helmholtz-muenchen.de/informationkommunikation/meldungen/news/index.html. That will reach app. 1800 employes at Helmholtz Zentrum München. During this process, the public relations office at HMGU will make a decision whether a press release will be sent to the media. Final Report February 2009/ Page 8 of 15

Did the project have particular relevance for other areas of society (cooperations with economy, effects on society, widespread impact, adult education etc.)? The Supporting Information published on the webpage of Cancer Research contains most of the mathematical details (in opposition to many mathematical oriented papers): http://cancerres.aacrjournals.org/content/suppl/2010/07/23/0008-5472.can-09-4392.dc1/sept_1.schollnberger.pdf It can be expected that this will help interested readers especially students to get an easier insight into the related rather sophisticated mathematics. Possible effects on cancer patient treatment are unclear at present. Final Report February 2009/ Page 9 of 15

3. Information on project participants not funded by the FWF funded by the FWF (project) co-workers number Personmonths non-scientific co-workers co-workers number Person - months non-scientific co-workers diploma students diploma students PhD students post-doctoral co-workers co-workers with Habilitation (professorial qualifications) PhD students 1 0.5* post-doctoral co-workers 1 0.25* co-workers with Habilitation (professorial qualifications) professors 1 0.25* professors * Project lasted from July 1 2009 until Sept. 30 2009. Numbers relate to the three coauthors Prof. Vineis, Dr. Beerenwinkel, and R. Hoogenveen. Final Report February 2009/ Page 10 of 15

4. Attachments List 1 1.a. scientific publications 3 with an indication of the status (published, in press, submitted, in preparation) 1.a.1. Peer-reviewed publications (journals, contribution to anthologies, working papers, proceedings etc.) Published: Schöllnberger H, Beerenwinkel N, Hoogenveen R, Vineis P. Cell selection as driving force in lung and colon carcinogenesis. Cancer Research 70: 6797-6803, 2010. All content of AACR journals is made free to anyone to read or use 12 months after its original publication at www.aacrjournals.org: 1.a.2. Non peer-reviewed publications (journals, contribution to anthologies research reports, working papers, proceedings, etc.) Published abstract: Mathematische Modelle nichtlinearer Dosiswirkungsbeziehungen für die strahleninduzierte Karzinogenese. 5. Biophysikalische Arbeitstagung 2010. 10. - 12. Juni 2010 in Bad Schlema. Biologische Wirkungen niedriger Strahlendosen - Natürliche Strahlenexposition, Radon-Balneotherapie und Strahlenschutz. RADIZ- Informationsheft Nr. 32/2010. Published proceedings: H. Schöllnberger. Mathematische Modelle nichtlinearer Dosiswirkungsbeziehungen für die strahleninduzierte Karzinogenese. Tagungsband der 5. Biophysikalischen Arbeitstagung 2010. 10. - 12. Juni 2010 in Bad Schlema. Biologische Wirkungen niedriger Strahlendosen - Natürliche Strahlenexposition, Radon-Balneotherapie und Strahlenschutz. ISBN 3-9811258-7-8. 1.a.3. Stand-alone publications (monographies, anthologies) 1.b. publications for the general public and other publications such as films, exhibitions, preparation of a home page etc. with an indication of the status (published, submitted / in preparation) Finished: http://www2.sbg.ac.at/fwf2/publications.html Planned media contributions: see bullet 2.3 of this report. 3 The publication list must mention for each work: all authors; full title; series/journal title; year; volume; and page numbers. Furthermore, if publications are freely available in the internet, please add the URL of the publication. Final Report February 2009/ Page 11 of 15

List 2 project-related participation in international scientific conferences 2.1. Conference participations - invited lectures Mathematische Modelle nichtlinearer Dosiswirkungsbeziehungen für die strahleninduzierte Karzinogenese. 5. Biophysikalische Arbeitstagung 2010. 10. - 12. Juni 2010 in Bad Schlema. Biologische Wirkungen niedriger Strahlendosen - Natürliche Strahlenexposition, Radon-Balneotherapie und Strahlenschutz. 2.2. Conference participations - lectures 3. Conference participations - posters 2.4. Conference participations - other Published abstract: H. Schöllnberger. Mathematische Modelle nichtlinearer Dosiswirkungsbeziehungen für die strahleninduzierte Karzinogenese. 5. Biophysikalische Arbeitstagung 2010. 10. - 12. Juni 2010 in Bad Schlema. Biologische Wirkungen niedriger Strahlendosen - Natürliche Strahlenexposition, Radon -Balneotherapie und Strahlenschutz. RADIZ-Informationsheft Nr. 32/2010. Final Report February 2009/ Page 12 of 15

List 3 Development of collaborations Indication of the most important collaborations (maximum 5), that took place (initiated or continued) in collaboration please give the name of the collaboration partner (name, title, institution) and a few words about the scientific content. Please also assign one of the following categories to each collaboration: N E D Nature N (national); E (European); I (other international cooperation) Extent E1 low (e.g. no joint publications but mention in acknowledgements or similar); E2 medium (collaboration e.g. with occasional joint publications, exchange of materials or similar but no longer-term exchange of personnel); E3 high (extensive collaboration with mutual hosting of group members for research stays, regular joint publications etc.) Discipline D within the discipline T transdisciplinary N E D Collaboration partner / content of the collaboration E E2 T 1) Name: Paolo Vineis Title: Prof., MD, MPH, FFPH Institution: Imperial College London, School of Public Health Content: scientific exchanges by email related to epidemiology and modelling E E2 D/T 2) Name: Niko Beerenwinkel Title: Prof. Dr. Institution: ETH Zürich, Computational Biology Group Content: intense scientific exchanges by email/phone on aspects related to cancer modelling E E2 T 3) Name: Rudolf Hoogenveen Title: MS Institution: National Institute for Public Health and the Environment (RIVM) Content: scientific exchanges by email on mathematical issues 4) Name: Title: Institution: Content: 5) Name: Title: Institution: Content: Note: general scientific contacts and occasional meetings should not be considered as collaborations in the above sense. Final Report February 2009/ Page 13 of 15

List 4 Habilitations (professorial qualifications) / PhD theses / diploma theses with an indication of the status (in progress / completed) 4.1. Professorial Qualifications 4.2. PhD Theses 4.3. Diploma Theses Final Report February 2009/ Page 14 of 15

List 5 Effects of the project outside the scientific field (where appropriate) Sections of the list: 5.1. Organization of scientific events 5.2. Particular honours, prizes etc. 5.3. Information on results relevant to commercial applications 5.4. Other effects beyond the scientific field Possible effects on cancer patient treatment are unclear at present. 5.5. Relevance of the project in the organization of the relevant scientific discipline Contributions to image building, establishment of priority areas, role in performance-related contracts or similar. Since the publication of the study in Cancer Research, Dr. Schöllnberger had related discussions with Drs. Georg Luebeck and Mark Little, National Cancer Institute. Both stated that it would be useful to study the question of the relative importance of promotion/selection versus mutation induction with the stochastic Multistage Clonal Expansion (MSCE) carcinogenesis model (Luebeck EG, Moolgavkar SH. Proc Natl Acad Sci USA. 99: 15095-15100, 2002). The MSCE model is an important recent extension of the MVK model and can fit certain epidemiological data that the MVK model cannot explain. According to Dr. Luebeck, the question related to the relative importance of cell selection versus mutation induction needs to be investigated looking at a bigger picture beyond the MVK model. That way the project has pointed out a new scientific avenue for the investigation of essential biological mechanisms in the formation of cancer cells in human solid tumours. List 6. Applications for follow-up projects with an indication of the status (submitted / approved) and the funding organization. 6.1 Applications for follow-up projects (FWF projects) (with an indication of the project type, e.g. stand-alone project, NFN, SFB, WK, fellowship, contribution to a stand-alone publication) 6.2 Applications for follow-up projects (Other national projects) (e. g. FFG, CD Laboratory, a K-plus Centre, funding from the Austrian National Bank, the Federal Government, the provincial government or similar) 6.3 Applications for follow-up projects (International projects) (eg. ERA project, ESF) Final Report February 2009/ Page 15 of 15