Apoptosis Destruction of Neoplastic Cells
Apoptosis Destruction of Neoplastic Cells Induced by the Free-Radical Transformation of Cerebrosides Into Ceramides at Photodynamic Therapy
Tech Area / Field
- BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
- INF-SOF/Software/Information and Communications
- INS-MEA/Measuring Instruments/Instrumentation
- MED-DIS/Disease Surveillance/Medicine
- MED-DRG/Drug Discovery/Medicine
- PHY-OPL/Optics and Lasers/Physics
3 Approved without Funding
B.I. Stepanov Institute of Physics, Belarus, Minsk
- Institute of Bioorganic Chemistry, Belarus, Minsk
- Universität Hamburg / University Hospital Hamburg-Eppendorf / Martini-Klinik am UKE, Germany, Hamburg\nUniversitat Leipzig / Institut fuer Medizinische Physik und Biophysik, Germany, Leipzig\nUniversite Montpellier II / Institut Charles Gerhardt, France, Montpellier
Project summaryThe purpose of the Project
The purpose of the Project is to develop a novel approach that will allow one to accelerate the destruction of malignant cells by apoptosis.
The State of the Art
The treatment of cancer is a worldwide problem. Among various methods, photodynamic therapy (PDT) is the most powerful method of treatment for cancer and certain non-cancerous diseases that generally characterized by overgrowth of unwanted or abnormal cells. In the vast majority of applications, the primary role of PDT is to kill unwanted cells. Although PDT can produce apoptosis or necrosis, or combination of the two mechanisms, in many clinical cases PDT is highly efficient in inducing necrosis. Also it is rather desirable that at PDT the destruction of malignant cells takes place mainly through the mechanism of apoptosis. This essentially reduces the intoxication of the tissue adjacent to the tumor and the development of inflammatory processes in an organism. Moreover, the efficient induction of apoptosis of tumor cells by PDT may help to overcome or bypass mechanisms of apoptotic resistance which develop in response to chemotherapeutic drugs and ionizing radiation action. It is unlikely that there is a universal mechanism for the response of cells to PDT. The responses vary with the cell type and its genetic and metabolic potential, as well as the photosensitizer and its sub-cellular localization. Presently more than dozen of synthetic sensitizers (some of them are used in clinical practice), for which both ways of tumor cells destruction are typical, are investigated. In the case of using 5-aminolevulinic acid and its esters for PDT, endogenous photosensitizer protoporphyrin IX is synthesized only in mitochondria of tumor cells causing their destruction mainly by apoptosis. In spite of a great number of papers published on this problem, molecular mechanism of cell destruction at PDT have not been fully investigated, and the results obtained by different authors are often contradictory. In a number of papers on the different PDT models it is shown that ceramides, which are formed under the action of sphingomyelinases on the sphingomyelin, play an important role in apoptosis induction. Recently the Project authors have established that ceramide may be formed in accordance with the reaction of free-radical fragmentation of with glycosphingolipids. The discovery of such a way of ceramide formation may become the essential chain in apoptosis induction mechanism. It allows one to intensify the PDT apoptotic effects by local or systematic injection of 5- aminolevulinic acid and its esters jointly with glycosphingolipids (cerebrosides). The fulfillment of the present Project also has great practical significance as a scientific basis of the development and creation of new effective drugs for tumors PDT, the result of which is cell destruction by apoptosis.
Impact of the Project on the Progress in the Field
The new basic knowledge about mechanism of free-radical transformation of with glycosphingolipids and product properties will be obtained. New data about the PDT dynamics determined by the ceramides concentration in culture cells will be gained. A correlation between the accumulation of protoporphyrin IX and ceramide and apoptosis destruction of cells in tumor tissues will be established. New data on photoinduced second messengers formation obtained at the Project fulfillment could be useful for cell and molecular biology and molecular pathology. The developed new methodology will be applied to clinical oncology and dermatology. The results obtained will allow developing new effective drugs inducing apoptotic cells destruction by PDT.
Competence of Project Participants
Twenty eight regular participants will take part in the Project. The scientific team involves 5 Doctors of Sciences in Physics and Mathematics, 2 Doctor of Sciences in Chemistry, 1 Doctor of Sciences in Medicine, 10 Candidates of Sciences in Physics and Mathematics, Chemistry, Medicine and Biology and 10 persons without scientific title but with a higher education. The majority of them are specialists in medicine, biology, chemistry and laser-scanning confocal microscopy and laser physics. Many of project participants were involved in the research programs of the military and industrial complex of the former USSR in the field of nuclear and biological weapon and missiles, as well as in researches connected with the laser-based navigation system of aircrafts. A number of participants have expertise in surgical, oncological and gastrointestinal pathology, immunohistochemistry, in the area of photobiology, and lipids biochemistry. They have accumulated a big experience on the chemical synthesis of lipids, 5-ALA and its derivatives, receiving and investigation of liposome, including medicinal junctions, physico-chemical and mass-spectrometric analysis of lipids and products of their free-radical transformations. 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 nuclear weapon and laser application.
Meeting ISTC Goals and Objectives
The Project fully meets the objectives of the ISTC on providing an opportunity for weapons scientists, engaged into the Project, to redirect their skills and experience to the peaceful activity. Foreign collaborators will be involved in the development of project proposal and working plan, exchange of information, consultations, reviewing of technical reports and intermediate documentation, external audit and carrying out joint seminars.
The following forms of cooperation with the Partner collaborators are planned:
- systematic informational exchange during the Project fulfillment and comments to the technical reports (quarter, annual, final), presented by the participants of the Project in ISTC;
- participation in technical monitoring of the Project fulfillment, realization of annual joint seminars;
- providing of the material aid (equipment, chemical compounds and experimental rooms);
- checking-out of the results obtained during the project fulfillment, preparation of the joint articles, reports and patents.
Expected Results and their Application
During the realization of 3 tasks of the project we are going to get the following results:
- to establish the laser-induced mechanism of enhancement of ceramides accumulation in tumor tissues according to the revealed by the Project authors of a new reaction of free-radical fragmentation of glycosphingolipids;
- to estimate PDT dynamics on the apoptosic pathway and the content of ceramides in the culture of tumor cells incubated with 5-aminolevulinic acid and cerebrosides;
- to establish the correlation between the protoporphyrin IX and ceramide accumulation in cells, and survival of cells in tumor tissues;
- to elaborate the fundamental methodical approach aimed on the laboratory animals’ tumor cells destruction on the apoptosic pathway at PDT treatment.
The received complex of results will allow one to develop and create new effective drugs for PDT that will cause the destruction of tumor tissues by apoptosis.
Scope of Activities
The Project duration will be 3 years. The total amount of efforts will be 8310 person-days. Within the scope of the Project, it is planned to develop a model of cell membrane, to establish physico-chemical liposome characteristics including cerebrosides and photosensitizers, to receive new knowledge about mechanism of photosensibilized free-radical transformation of glycosphingolipids and product properties. It is necessary to obtain new data about protoporphyrin IX biosynthesis and to estimate PDT dynamics and the content of ceramide in the culture of tumor cells incubated with 5-aminolevulinic acid and cerebrosides. In addition, it is planned to establish the correlation between the accumulation of protoporphyrin IX and ceramide and survival of cells in tumor tissues.
Role of Foreign Collaborators
The following forms of cooperation with the foreign Collaborators are planned:
- systematic information exchange in the course of the Project implementation;
- participation in the technical monitoring of the Project activities;
- cross-checks of the results obtained in the course of the Project implementation;
- participation in experimental data analysis and preparation of joint scientific publications.
Technical Approach and Methodology
The technical approach and the methodology are based on the many years experience of the Project participants in the areas of medicine, biology, chemistry and scanning-laser microscopy accompanied by Forster’s resonance energy transfer (FRET) and Fluorescence Life-time Imaging System (FLIM). Scanning-laser confocal microscopy is the best and most accurate method of investigating the dynamics of biological processes inside cells and tissues providing 3D resolution.
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