Abstract 74P
Background
Mitochondria (Mi) are fundamentally implicated in cancer biology, including initiation, growth, metastasis, relapse, and acquired drug resistance. Mi are considered as the target organelles for therapeutic strategies of several cancers including CCA. The project aims to dissect the novel MiPa, to identify all molecular players, to functional characterize their role in CCA, to identify novel targeted therapies, to develop a gene signature exhibiting consistent prognostic power and predictive value as potential biomarker.
Methods
HTS has been performed by using Prestwick Chemical Library (Prestwick) and siGENOME Druggable-Genome-Library (Dharmacon). CASP9 activity has measured using the ApoAlert CASP9 assay (Clontech). Cell viability was monitored by CellTiter-Blue assay (Promega). The patient cohort consists of 100 patients retrospectively identified by Univeristy of Modena, IT. Lentiviral-based shRNA and CRISPR/Cas9 systems were utilized to establish stable gene knockdown and knockout model. FFPE-RNA extraction, RNAseq, WES and bioinformatic analyses will beperformed using protocols and pipeline available at TIGEM (2).
Results
Molecular players of the novel MiPa, which triggers APAF1-independent CASP-9 activation have been identified using a proteomic approach combined to siRNA library screening. HTS allowed to identify compounds targeting the novel MiPa. Two compounds were found to inhibit tumour growth in vitro and their anti-tumour efficacy has been assessed. Experiments in HuCCT-1, KMCH, CCLP, SW1, EGI, TFK1 cell lines revealed their effect on autophagy inhibition and apoptosis and correlated the MiPa pathway with drug treatment response and drug resistance. Whole-genome analysis is ongoing to profile the novel MiPa gene signature in cell lines treated and not with standard therapeutic approach and in a cohort of 100 CCA patients.
Conclusions
A novel MiPa has been identified regulating the apoptosis in CCA. Implication of the novel MiPa in drug resistance and sensitivity to classical therapeutic treatment has been assessed. Whole-transcriptome and exome analysis are ongoing to evaluate the clinical value of the MiPa gene signature in CCA patients. References: 1. Indrieri, A., et al., EMBO Mol Med, 2013. 2. Cacchiarelli D, et al., Cell. 2015.
Clinical trial identification
Editorial acknowledgement
Legal entity responsible for the study
The authors.
Funding
TIGEM - Telethon Institute of Genetics and Medicine.
Disclosure
All authors have declared no conflicts of interest.