Title of project: Screening for novel functions of endocytic and autophagic proteins in the regulation of gene expression, cell growth and carcinogenesis.
Person responsible for project (IIMCB): Iwona Pilecka, PhD
Norwegian partner: Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo (IBMS)
Person responsible for project (IBMS): Anne Simonsen, PhD, Associate professor
Summary of the project:
The Project, proposed by two research groups of the International Institute of Molecular and Cell Biology (IIMCB) in Warsaw and of the Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, belongs to the field of pathogenesis and genetics of cancerous diseases. It aims to identify candidate genes potentially involved in tumorigenesis, using state-of-the-art molecular screening approaches.
The Project is based on novel data indicating that hyperproliferative conditions underlying cancer can result from the perturbation of endocytosis and autophagy, two cellular processes involving lysosomal organelles. Endocytosis is a process of vesicular transport in cells which ensures uptake of extracellular molecules (e.g. nutrients or growth factors) and their subsequent degradation in lysosomes or recycling back to the plasma membrane. Autophagy is a lysosomal pathway for bulk degradation of cytoplasmic contents and whole organelles which complements apoptosis. In recent years, several endocytic and autophagic proteins, initially characterized in the context of vesicular transport, were reported to act as tumor suppressors or function directly as regulators of transcription in the cell nucleus. However, no systematic studies were undertaken to search for such alternative functions of endocytic/autophagic proteins or to elucidate the underlying molecular mechanisms, and our Project aims to fill this gap by an innovative and comprehensive approach.
We thus propose to establish a screening platform for cell-based assays and perform a screen for an initial set of endocytic and autophagic proteins for their potential roles in carcinogenesis by measuring their involvement in signal transduction, transcription and cell proliferation, by RNA interference-mediated knockdown. The Project activities are divided into three main tasks within a total timeframe of 24 months. We will generate libraries of siRNA reagents for the selected endocytic/autophagic genes (Task I). These reagents will be used in four primary screens, using automated microscopy-, fluorescence- and luminescence-based readouts in a middle-throughput scale format (Task II), followed by validation of positives in two secondary assays (Task III). Upon Project completion, we expect to have identified candidate genes potentially involved in tumorigenesis, for future testing in clinical samples.
Thus the Project should contribute to conceptual advance of knowledge on molecular mechanisms of cancer but equally important will be an establishment of a largely automated screening platform at IIMCB as a basis for further high-throughput, eventually genome-wide, screening projects. In a long term this will contribute to introducing novel technologies and systems biology approaches into the field of cancer research in Poland.