NKUA-CRG researchers and academic members will have via INsPiRE the opportunity to fulfil their vision, acquire innovative knowledge and develop further the three main research axes.
- a. Tumour promoting factors and anti-tumour barriers.
Considering the enormous tumour complexity it is planned to study the molecular and biochemical characteristics of specific parts or cell types of human tumours derived from biopsies. New immunolocalization methods at the CLSM and EM level will be applied to visualize DNA damage and to map the topological distribution of the molecules under study in normal and cancer human cells. The functional implication of these molecules in the various types of cellular death and cell cycle regulation will be further investigated, while as a number of preliminary data indicate an existing cross-talk between the DNA Damage Response (DDR) machinery and the proteasome proteolytic system, a particular effort will be invested in the understanding of the possible links between these two vital cellular mechanisms. Finally, the implication of the studied molecules in the DDR response and EMT or in the regulation of the insulin pathway will be further evaluated at in vitro and in vivo models.
- b. Identification and functional analysis of tumour biomarkers.
Further investigation and validation of the identified cancer-related biomarkers (and their isoforms) at the mRNA and protein level in normal human cells and/or tissues, in a significant number of tumours, chemoresistant cell lines and blood samples as well as during immunomodulation and response to immunotherapy is scheduled. Also it is planned to identify novel molecular entities (e.g. miRNAs) that regulate the expression profile of the studied cancer related biomarkers. The combination of these molecules with the stabled biomarkers into multiparametric models will provide important diagnostic and prognostic information for breast, prostate and colon cancer. Transgene mediated protein overexpression and mRNA knock-down via siRNA will be used to functionally analyze the identified biomarkers. These approaches will give important insight into the incomplete understanding of structural, functional and regulatory questions that remain unanswered for the studied genes.
- c. Cancer immunotherapy/Drug targeting of cancer-related signalling networks.
Considering that many sources of natural products remain largely untapped (especially those that relate to the beneficial effects of the Mediterranean diet) a high throughput screening of newly isolated compounds with suspected anti-cancer activity is planned. Several human cancer cell lines (sensitive or resistant to chemotherapeutic drugs) will be exposed to different concentrations of these compounds and high-throughput MTT assays will evaluate their cytostatic/cytotoxic effects. Selected agents that meet the criteria set will be optimized for new cycles of evaluation. New compounds or those with known kinase inhibitory activity (see above) will be tested for their ability to modulate cancer-related signalling networks, kinases, EMT or proteasome activities. In order to conclude the pre-clinical studies, this setting will be complemented with in vivo (animal) experiments (e.g. xenografts) using wide-ranging doses of the selected compound(s) to obtain preliminary efficacy, toxicity and pharmacokinetic information. Moreover, the mode of action of these compounds in optimizing immune cell stimulation and promoting adequate anti-tumour responses will be evaluated both in vitro and in vivo. Emphasis will be given to those compounds able to activate dendritic cells to further elicit both, CD8+ and CD4+ T cell and/or NK cell cytotoxic responses. Finally, to design specific inhibitors for various molecules of interest (e.g. CLU, Cdt1 or Cdc6) studies will be focused on acquiring information about their respective molecular crystal structure. Such tests, along with the discovery of new biological markers for response to drug therapy, or approaches to increase the compound’s efficacy (e.g. via siRNA-mediated knock-down of cytoprotective genes) will assist NKUA-CRG members and collaborating pharmaceutical companies in deciding whether a “drug candidate” has scientific merit for further development as an investigational new drug.