PRR Project

Summary

This position is for an Assistant Researcher of the scientific research career, in the scientific domain/area of Medical and Health Sciences-Basic Medicine-Human Genetics, to work in the Cancer Genetics Group of the IPO Porto Research Center, hosted by IPO Porto. The general objective of the Cancer Genetics Group is to study the interplay between inherited cancer predisposition and the pattern of acquired genetic alterations that give rise to cancer, in order to understand the mechanisms of tumor initiation and progression and their role in therapy response and resistance. The research group has a strong focus in prostate cancer research, especially to uncover the missing heritability behind predisposition to prostate cancer by combining exome sequencing and haplotype analyses in a population with strong founder effects. This strategy allowed us recently to identify two novel prostate cancer predisposition genes, namely PRUNE2 (Exome sequencing of affected duos and trios uncovers PRUNE2 as a novel prostate cancer predisposition gene, Pubmed ID: 36564567) and IPO4 ( IPO4 is a new DNA repair gene contributing to inherited prostate cancer predisposition; submitted).Considering this recent output of the Cancer Genetics Group, the Assistant Researcher to be hired is expected to play a central role in the following main lines of research:1. Building on the previous findings of pathogenic germline variants in the PRUNE2 gene in early-onset/familial prostate cancer patients, this project aims to dissect the independent roles of PRUNE2 and PCA3 in the function of the PRUNE2/PCA3/AR triad in prostate carcinogenesis. We will use a multidisciplinary approach, integrating computational transcriptomics, CRISPR/Cas9 gene editing, fluorescence microscopy and optimization of pharmacological targeting in cancer cells, namely to: identify dysregulated genes and pathways associated with PRUNE2 alterations, by performing whole-transcriptome analysis (RNAseq); evaluate the independent functions of PRUNE2 and PCA3 in prostate cells, by establishing single and double knockout models for PRUNE2 and PCA3 using  prostate-derived cell lines; evaluate the impact and phenotypic consequences of anti-androgen therapy on the PRUNE2/PCA3/AR triad, by using an androgen antagonist (Enzalutamide) and an agonist (DHT); and, investigate the effects on downstream molecular targets and oncogenic signaling pathways caused by depletion of PRUNE2 and/or PCA3, using western-blot.2. Considering our previous findings of pathogenic germline variants in the IPO4 gene in early-onset/familial prostate cancer patients, the involvement of IPO4 in the Fanconi Anemia (FA) pathway, and the recent approval of PARP inhibitors for the treatment of metastatic castration-resistant prostate cancer in patients carrying deleterious variants in DNA damage repair genes, we aim to clarify the potential clinical relevance of IPO4 variants. Preliminary data using CRISPR/Cas9-engineered knockout (KO) models showed increased levels of chromosome instability (CIN) in IPO4 KO cells upon treatment with diepoxybutane, suggesting a role in DNA damage repair. This finding was linked to deficient recruitment of FANCD2 to the DNA damage sites in the IPO4 KO cells, providing a rational mechanism for CIN. To explore the potential therapeutic implications of IPO4 variants in prostate cancer patients with advanced disease, we will start by using IPO4 KO cells to evaluate the sensitivity to PARP inhibitors after mitomycin C sensitization, using zebrafish xenografts. Additionally, to demonstrate the clinical relevance of the findings, we will screen a series of metastatic prostate cancer patients for germline mutations in IPO4 using either tumor or cell-free DNA, using targeted next generation sequencing (NGS). Altogether, this work will support IPO4 as a new prostate cancer risk gene and establish IPO4 as a player in the maintenance of genomic integrity through its involvement in the FA pathway.In order to be able to play a central role in these main lines of translational research of the Cancer Genetics Group, the Assistant Researcher to be hired should have a track record in prostate cancer inherited predisposition combined with strong molecular biology skills that are required to move beyond the state-of-the-art genomics’ research, namely: PhD in Biomedical Sciences or related areas, combined with training in Biotechnology Engineering and Molecular Oncology, with more than 6 years of post-doctoral experience; strong publication record in high profile, peer reviewed international scientific journals, including several as first author; documented experience in cancer genetics, with emphasis on inherited prostate cancer predisposition; documented experience in NGS, from wet -lab to data analysis and interpretation; experience in cell-based gene silencing/ de novo expression and functional phenotypic assays; and training in bioinformatics, including genome-wide gene expression analysis.