Project Funding Details

NFY is a pioneer transcription factor that in physiological conditions drives the transcription of a plethora of cell-cycle regulatory genes, thus acting as a master regulator of proliferation. It is a heterotrimer composed of NFYA, NFYB, and NFYC subunits, of which NFYA mediates sequence-specific binding to CCAAT-boxes on the DNA. Consistent with the fact that uncontrolled proliferation is a distinctive hallmark of cancer, recent studies show that NFYA subunit is upregulated in most human tumors, where it sustains cell growth/survival and metabolism. Although different layers of regulation have been shown to control the expression of NFY, post-transcriptional mechanisms have been poorly investigated, especially with regard to the role of noncoding RNAs. In the last 20 years, microRNAs (miRNAs) and, more recently, long noncoding RNAs (lncRNAs) have been increasingly recognized as regulators of pivotal biological processes, with their aberrant activity being implicated in tumorigenesis. It is hence conceivable that dysregulated noncoding RNA function may contribute to aberrant NFY expression in cancer. The overarching goal of the proposal is to help elucidate the relevance of noncoding RNA-mediated regulation of NFY in human cancer. The working model will be represented by prostate cancer, on which we have long-standing preclinical experience and availability of patients' material. Specific aims are: 1) The dissection of NFYA mirnome 2) The first functional characterization of the peculiar antisense lncRNA transcribed from NFYC bidirectional promoter: NFYC-AS1 3) The definition of the translational relevance of NFY noncoding-ome as prognostic biomarker as well as of the clinical utility of NFY noncoding-ome targeting to improve tumor response to radiotherapy. We will use an integrated approach involving cell, molecular and computational biology, articulated as follows: 1) Identification of miRNAs (including isomirs) targeting NFYA, upon accurate re-annotation of its 3'UTR, and genetic manipulation of miRNA responsive elements to disrupt miRNA-mediated regulation; 2) gain- and loss-of-function approaches and up-to-date molecular biology techniques to dissect NFYC-AS1 role; 3) analysis of clinical samples to define the prognostic role of NFY noncoding-ome components and in vitro/in vivo assays to assess the impact of NFY noncoding-ome targeting on tumor radiosensitivity. The project should lead to the identification of NFYA-targeting miRNAs and to the biological characterization of how loss of miRNA-mediated regulation of NFYA may impact on cell transformation and acquisition of tumor features; the definition of NFYC-AS1 transcript structure, subcellular localization, role and mechanism of action in cancer; and the definition of the prognostic value of NFY noncoding-ome and of the clinical utility of NFY noncoding-ome targeting to improve tumor response to radiotherapy. Overall, the results obtained in the 5-yr time frame should lead to i) an overall more exhaustive characterization of regulatory pathways that, when altered, can favor prostate cancer development and progression, ii) identify biomarkers able to predict tumor relapse prior to biochemical evidence and iii) furnish the biological rationale for the development of novel therapeutic opportunities for aggressive prostate cancer based on NFY noncoding-ome targeting combined to radiotherapy, which could help treatment optimization and improve patients' survival in the medium-long term.