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Dolfini Diletta
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Roberto Mantovani |
Bioinformatic Identification of key transcription factors in tumors. |
The synergy between Transcription Factors regulates gene expression. TFs are pivotal in the control of cell growth: alteration of their structure or e ... The synergy between Transcription Factors regulates gene expression. TFs are pivotal in the control of cell growth: alteration of their structure or expression leads to tumorigenesis. NF-Y is a "pioneer" TF binding the CCAAT box, a crucial element of promoters. It is a trimer made of histone fold domain dimer NF-YB/NF-YC and the sequence-specific regulatory NF-YA. NF-YA exists in two major isoforms, “short” and “long”. Independent evidence suggest that NF-Y plays a relevant role in cancer progression. (i) Profilings of tumor vs normal cells show enrichment of CCAAT in cancer "signatures" genes. (ii) A genome-wide connection to a plethora of oncogenic and growth controlling TFs. (iii) Crucial genes in metabolic pathways altered in cancer cells are controlled by NF-Y. NF-YAs correlates with a poor prognosis in ovarian cancer. In the last years, we define NF-YA isoforms role in epithelial cancers -breast , lung, head and neck: we found that NF-YAs levels correlate to a proliferative signature, while NF-YAl is associated to mesenchymal cellular identity.
Tumors rely on CCAAT-binding to activate a significant number of "cancer" genes. An upregulation of NF-YA could corroborate the malignant transformation of cells and a switch between long and short isoforms could push an invasive behavior concurring to formation of metastases. The critical questions are: could NF-YAs subvert normal gene expression and mediate transformation? Could NF-YA isoforms switch be a marker of transformation and aggressiveness of tumor cells?
We aim at obtaining a fine molecular knowledge of NF-YA isoform switching during transformation of cells in cancers. We will perform in-depth analysis of available tumor gene expression data to characterize the switching of NF-YA and in general of TFs isoforms in these tumors. This project will clarify the role of NF-YA and TF isoforms in defining epithelial or mesenchymal cancer signatures. |
- Molecular Biotechnology and Bioinformatics (MB&B)
- Molecular Biology of the Cell (MBC)
- Bioinformatics for Computational Genomics (BCG)
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September2022 |
1 |
12/04/2022 |
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Mantovani Roberto
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Diletta Dolfini |
Stem cell, Alternative Splicing |
The Project is focused on the role of alternative splicing in early development, notably in the formation of the mesoderm. Alternative splicing produ ... The Project is focused on the role of alternative splicing in early development, notably in the formation of the mesoderm. Alternative splicing produces the two major isoforms of NF-YA (the "long" NF-YAl and "short" NF-YAs, without exon-3 sequences)- NF-YA is the regulatory subunit of the trimeric Transcription Factor NF-Y. NF-YAl expression correlates with the activation of early mesoderm differentiation. We ablated exon-3 of NF-YA in mouse Embryonic Stem Cells using genome editing: we observed a loss of mesendoderm population during differentiation to Embryoid Bodies. The result was validated by qRT-PCRs, and finally confirmed by single cell RNA-seq. The machanism apprears to be related to activation of a casacde of mesodermal master transcriptional regulators (Brachyury, EOMES, SNAI2).
The Project aims at identifying the proteins mediating the alternative splicing switch from NF-YAs (Stem/Ectoderm) to NF-YAl (Mesoderm), which is fundamental in turning on the mesodermal program.
Genome editing, RNAi, single cell analysis, differentiation of stem cells.
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- Biologia Applicata alla Ricerca Biomedica (BARB)
- Molecular Biotechnology and Bioinformatics (MB&B)
- Molecular Biology of the Cell (MBC)
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Ottobre 2022 |
1 |
12/04/2022 |
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Mantovani Roberto
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Diletta Dolfini |
Cancer, Alternative splicing |
The Project is focused on the role of alternative splicing in cancer development, notably breast and gastric. Alternative splicing produces the two m ... The Project is focused on the role of alternative splicing in cancer development, notably breast and gastric. Alternative splicing produces the two major isoforms of NF-YA (NF-YAl and NF-YAs, without exon-3 sequences), the regulatory subunit of the trimeric Transcription Factor NF-Y. NF-YA is overexpressed in most epithelial cancers and NF-YAl expression correlates with the presence of aggressive cells that underwent EMT (Epithelial Mesenchymal Transition). We ablated exon-3 of NF-YA in two breast cancer cell lines using genome editing and observed a loss of aggressive features (migration and methastasis) in vitro and in vivo. The mechanism appears to be related to activation of EMT transcription Factors. The issue is now how splicing of NF-YA isoforms is regulated.
The Project aims at identifying the proteins involved in the alternative splicing switch from NF-YAs (epithelial) to NF-YAl (EMT). Candidate factors will be assessed by genetic (Genome editing) and molecular biology (RNAi) means. |
- Biologia Applicata alla Ricerca Biomedica (BARB)
- Molecular Biotechnology and Bioinformatics (MB&B)
- Molecular Biology of the Cell (MBC)
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Ottobre 2022 |
1 |
12/04/2022 |