State of the art in the field of the project. The treatment of cancer is a worldwide problem. The understanding of mechanisms of tumor growth and progression is critical for novel anticancer drug development. There is at present no chemotherapeutic agent that suppresses cell migration, although such a drug could certainly improve survival because of inhibition of metastasis.

Colorectal cancer is a major global health problem with more than a million new cases diagnosed worldwide every year. Moreover colorectal cancer is a good model for studying tumor invasiveness because of the easily detectable invasive front. Thus, the results obtained from colorectal cancer studies could be fundamental for all malignancies of epithelial origin. That is why we have chosen colorectal cancer for our study.

The invasive tumor behavior underlies increased cell motility caused by changes in cytoskeletal organization and alteration of contacts with the extra-cellular matrix (ECM). Our preliminary results indicate that certain cytoskeleton changes should be critical in acquiring migrating phenotype. It is also well known that extracellular remodeling is significant in cell migration. The same ECM remodeling occurs in both cancer and inflammatory diseases. We suggest that cytoskeletal changes and ECM remodeling are closely related. We are going to reveal if there is any parallel between these two processes in the invasive front of colorectal cancer.

It is well known that many of the molecular events associated with tumor growth also occur in inflammation. Considering this fact, it is important to define the cytoskeletal changes on the invasion border. Thus, we are going to compare the results obtained on cancers with those in inflammatory bowel diseases (IBD), i.e. ulcerative colitis and Crohn’s disease.

The main problem in understanding cell migration is that in vitro results concerning this phenomenon are of limited value in vivo. We are going to provide our research only in vivo (ex vivo) on histological material obtained after surgery or biopsy. Thus, the results we obtain will reflect the real state of cytoskeletal and ECM molecules in a real patient

Impact of the proposed project on the progress in the field of the development.

• The obtained data about microtubule rearrangement in cancer cells and in inflamed regions will extend our knowledge about cell migration and cancer invasion. Little is presently known about this aspect of cell migration. Moreover, these data will be accompanied by new information about changes in ECM composition associated with changes in tubulin isotype expression.
• The knowledge gained during the project about the role of changes in tubulin expression in cancer cells and in inflamed regions will probably open a new area for anti-cancer drug development.
• The application of mathematical approach based on “fuzzy logic” in computerized image analyses will give an opportunity to develop very novel and unique software of wide practical significance.

The pathological group has expertise in surgical, oncological and gastrointestinal pathology, immunohistochemistry and confocal microscopy. This group includes 2 Dr. Sci and 3 PhDs. The members of this team have more than 100 articles and scientific works. The former military specialists from this group have experience in the field of medical and biological consequences of nuclear and biological weapon applications.

The clinical group includes clinical oncologists and gastroenterologists. They have more than 100 articles and scientific works on gastroinstestinal cancers and IBD problems. This group includes 1 Dr. Sci and 2 PhDs.

The confocal group consists mainly of former weapon scientists. They have a great deal of experience in optics, lasers and their military applications. This group includes 2 Dr. Sci. This group is equipped by very unique confocal microscope.

The image analyses group also contains former weapon scientists. Their weapon specialization is image analysis for satellite and missile purposes and programming for military aims. They have a lot of experience in image analysis for military and medical purposes, and in creation of algorithmes based on fuzzy and possibilistic clustering. This group includes 1 Dr. Sci and 2 PhDs.

Expected results. During the realization of 3 tasks of the project we expect to obtain the following results:

1. Data about the pattern of cytoskeletal changes in the invading cancer cells and in epithelial cells in inflammatory sites.
2. Characterization of the relationship between cytoskeletal rearrangement and ECM changes in cancer and inflamed colon tissue.
3. New algorithm of image analysis of immunohistochemical slides.

Scientific significance.

1. The new basic knowledge about cytoskeleton changes in epithelial cells in cancer and inflamed tissue will be obtained. The role of the cytoskeleton in cell migration is presently very poorly understood.
2. The new data about the relationship between cytoskeletal rearrangement and ECM changes will be gained.
3. The existing knowledge about cancer metastasis will be significantly extended.

1. New molecular targets for anti-cancer drug development
2. New markers for prediction of colorectal cancer progression based on cytoskeleton and ECM changes will be proposed and grounded
3. New image analysis software for quantitative immunohistochemistry will be created

Realization of ISTC aims and tasks.

The main part of project participants is presented by scientists and specialists participated in the programs of the military and industrial complex of the former USSR in the fields of:

• security facilities from weapons of mass destruction, particularly nuclear and biological weapon
• protection facilities of defense technologies from high-precision radio-controlled weapon
• optic pointing systems
• powerful lasers

Thus the project contributes for the next ISTC aims and tasks:

• This project provides former weapon scientists, who are going to participate in the project, the opportunity to redirect their knowledge and skills to peaceful activities, particularly, to cancer struggling.
• The project represents a basic and applied research in the field of oncology, cell and molecular biology, computer science
• The project supports the integration of scientists (medical scientists, physicists and mathematicians, weapon scientists), who are going to participate in the project, into the international scientific community by communicating with foreign collaborators and taking part in international conferences and seminars in the field of oncology, gastroenterology, image analyses, microscopy, cell and molecular biology and drug discovery.
• The project contributes to the solution of a worldwide problem of cancer.
• The project assists the transition to market-based economy by creating commercial software which will be demanded by medical centers, clinics and universities for practical and scientific needs.

Planned role of foreign collaborators. Foreign collaborators have been already involved in the development of project proposal and working plan. They will also participate in the exchanging of information, consultations, reviewing of technical reports and intermediate documentation, external audit and joint seminars.

Technical approach and methodology. We are going to provide our research on histological material obtained after surgery or biopsy. In comparison with an experimental approach this one allows us to examine the real state of cytoskeletal and ECM protein expression in real patients The expression of cytoskeletal and ECM proteins will be studied by immunohistochemistry and immunofluorescence detected by confocal microscopy accompanied by Forster’s resonance energy transfer (FRET). Confocal microscopy is the best and most accurate method of colocalization of two or more proteins in one cell or in one region. FRET will give us an opportunity to measure the degree of interactions between studied molecules of cytoskeleton in the cells. The developed image analysis software will provide us with the powerful tool for quantitative evaluation of immunohistochemistry.