Research objectives and content
The physiological importance of apoptosis is underlined by the existence of pathologies coupled to a dysregulation of apoptosis. Excessive apoptosis in involved in acute diseases (stroke, infarction, fulminant hepatitis, septic shock etc.) as well as ii chronic pathologies (neurodegenerative diseases, AIDS). Abnormal resistance to apoptosis accounts for hyperplasias and cancerogenesis. As a consequence, the exploration of the subcellular and molecular mechanisms of apoptosis, may yield important clues for the treatment of numerous pathologies. Our main objeetives are (i) the amelioration of methods for the detection of the of early stage of apoptosis; (ii) the exploration of the molecular and subcellular mechanisms of apoptosis induced via a pathway also involved in AIDS- associated Iymphocyte depletion, the so-called Fas-pathway: (iii) the determination of the i mechanisms by which the onco-protein Bcl-2 suppresses apoptosis, (iv) the development of strategies for the pharmacological modulation of apoptosis. Four major methodological approaches will be employed to achieve these goals: (i) cytofluorometric analysis and purification of cells and isolated organelles (nuclei, mitochondria) after labelling for antigen expression (CDs, Fas, Bc1-2, Bax, Bcl-X etc.), mitochondrial parameters DELTA-PHYm, superoxide anion production, cardiolipin content); glutathione levels, cytosolic or mitochondrial calcium, plasma membrane organisation, activation of specific proteases from the ICE/CPP32 family etc.; (ii) definition of the conditions (proteases, second messengers, Bcl-2 expression etc.) in which isolated mitochondria and/or cytosolic factors induce nuclear apoptosis in a "cell-free" system of apoptosis; (iv) pharmacological studies using specific protease inhibitors and mitochondrion-targeted drugs to modulate apoptosis in this cell-free system and in intact cells. We anticipate that this project will lay the theoretical grounds for pharmacological interventions on apoptosis.
Training content (objective, benefit and expected impact)
The host laboratory provides training: (i) in the application of multiparameter cytofluorometry to the detection of early
apoptosis-associated changes in cells and organelles; (ii) in the sorting of cells representing a defined stage of the apoptotic process; (iii) in the purification and analysis of subcellular compartments; (iv) in the characterization of cell-free systems of apoptosis; (iv) in the pharmacological evaluation of apoptosis modulators in vitro and in vivo. The candidate, the host institution, as well as the laboratory to which the candidate will return to in Spain will have the opportunity to engage in mutually rewarding cooperative research projects.
Links with industry / industrial relevance (22)
Dr. Guido Kroemer's group has tight connection with two institutions which are sponsored by the pharmaceutical industry, yet are formally independent: Institut Scientifique Roussel (Roussel-Hoechst) and the Leo Research Foundation (Upjohn-Pharmacia). In addition, Dr. Kroemer is independent consultant for Roussel-Uclaf. Since apoptosis control may yield clues for the treatment of cancer and AIDS, the pharmaceutical impact of this study cannot be denied.