MARTINA GENTILE

Dottoressa di ricerca

ciclo: XXXIV


supervisore: CLARA NERVI
relatore: CLARA NERVI
co-supervisore: ELISABETTA DE MARINIS

Titolo della tesi: Epigenetic role of microRNAs in normal and neoplastic hematopoiesis

Epigenetic signals and microRNA (miRNA) are key regulators of developmental cell programs, including hematopoietic cells lineage-specification and fate. Alteration of their function is at the basis of neoplastic transformation. miRNAs mainly mediate the post-transcriptional gene silencing of target mRNAs. However, miRNAs also bind complementary DNA sequences at specific gene sites and participate in the epigenetic regulation of chromatin structure. The general aim of the PhD research project was the identification of novel epigenetic mechanisms involving miRNAs and/or their targets and their relevance in normal and neoplastic hematopoiesis. To this end we: i) analysed the genomic seeding and DNA sequences bound by the myelopoiesis regulator miRNA-223 during the maturation/differentiation hematopoietic progenitors (HPCs); ii) addressed the epigenetic mechanisms of transcriptional regulation of target genes by miR-223; iii) studied the expression patterns and biological relevance of identified miR-223 bound genes in normal and leukemic hematopoietic cells; iv) identified and functionally validated new miRNAs and pathways that are relevant for myeloproliferative disorders. In chromatin isolated from myeloid cells undergoing granulocytic differentiation by retinoic acid (RA) treatment we identified at the whole genome level (ChIP-Seq) the promoter regions targeted by miR-223, presenting a spread overlap with activating (H3K4me3) and/or or repressing (H3K27me3) histone marks. Among the genomic loci bound by miR-223, we identified the Flotillin 1 (FLOT1) gene, which is associated with transcriptionally active H3K4me3 marks. FLOT1 encodes for a lipid-rafts associated protein, whose role in normal and neoplastic hematopoiesis is mostly unknown. We validated the expression levels of FLOT1 in myeloid cell lines and confirmed its induction by RA treatment, which is inhibited by miR-223 silencing. FLOT1 is highly induced in HPCs isolated from normal donors’ cord blood and peripheral blood undergoing granulocytic differentiation; its expression levels are significantly altered in bone marrow samples from acute myeloid leukemia (AML) patients. Overall, these data suggest FLOT1 as a transcriptional target of miR-223, playing a role in normal and leukemic differentiation. We also proceeded to the identification of miRNAs involved in the pathogenesis of Myeloproliferative disorders (MPNs), by using as a model system a novel CALR non-canonical mutation (del24), deleting 24nt of the 3’UTR mRNA proximal region, associated to increased erythropoiesis and a Polycytemia Vera (PV) phenotype. Bioinformatic analysis predicted that miR-1972 and miR-455-3p target sites are lost in CALR del24 mRNA, thus suggesting a role for these miRNAs in the differentiation program of myeloid progenitors and in the control of CALR expression levels. Indeed, we found a significant correlation between the expression levels of miR-1972 and CALR mRNA/ protein levels in myeloid cell lines, in HPCs undergoing unilineage maturation/differentiation and in a large series of MPNs patients. We addressed the biological relevance of CALR-3’UTR targeting by miR-1972 in a K562 CRISPR/CAS9 cell model carrying or not CALR del24. In wild type and mock K562 cells the inhibition of miR-1972 activity increased erythroid differentiation, whereas K562 clones with CALR del24 were not responsive, thus suggesting a role for the post-transcriptional CALR gene silencing by miR-1972 in the erythroid differentiation program of myeloid cells.

Produzione scientifica

11573/1711432 - 2024 - Nuclear miRNAs: Gene Regulation Activities
Billi, Monia; De Marinis, Elisabetta; Gentile, Martina; Nervi, Clara; Grignani, Francesco - 01g Articolo di rassegna (Review)
rivista: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (Basel: MDPI Center) pp. - - issn: 1422-0067 - wos: WOS:001245365800001 (2) - scopus: 2-s2.0-85195836823 (2)

11573/1710483 - 2024 - Nuclear miR-223 affects chromatin organization and regulates Flotillin-1 gene expression in myeloid progenitors
De Marinis, Elisabetta; Monia, Billi; Ianni, Alessandro; Gentile, Martina; Quattrocchi, Alberto; Zaza, Alessandra; Noguera Nelida, Ines; Borello, Ugo; Grignani, Francesco; Nervi, Clara - 04d Abstract in atti di convegno
rivista: MEDITERRANEAN JOURNAL OF HEMATOLOGY AND INFECTIOUS DISEASES (Roma : Istituto di Ematologia dell'Università Cattolica) pp. 19-20 - issn: 2035-3006 - wos: (0) - scopus: (0)
congresso: 8th Symposium on Acute Promyelocytic Leukemia (Rome, Italy)
libro: Mediterranean Journal of Hematology and Infectious Diseases - ()

11573/1710484 - 2023 - The epigenetic/transcriptional activity of nuclear mir-223 regulates flotillin-1 expression and function in myeloid differentiation
Billi, Monia; De Marinis, Elisabetta; Ianni, Alessandro; Gentile, Martina; Quattrocchi, Alberto; Zaza, Alessandra; Noguera Nelida, Ines; Borello, Ugo; Grignani, Francesco; Nervi, Clara - 04d Abstract in atti di convegno
congresso: 1st Epigenetics Society International Meeting "Epigenetics of Disease and Development" (Rome, Italy)
libro: 1st Epigenetics Society International Meeting “Epigenetics of Disease and Development” - (9782832512357)

11573/1692211 - 2023 - Biomarkers in acute myeloid leukemia. From state of the art in risk classification to future challenges of RNA editing as disease predictor and therapy target
Quattrocchi, Alberto; Cappelli, Luca Vincenzo; De Simone, Giovanna; De Marinis, Elisabetta; Gentile, Martina; Gasperi, Tecla; Pulsoni, Alessandro; Ascenzi, Paolo; Nervi, Clara - 01g Articolo di rassegna (Review)
rivista: ASPECTS OF MOLECULAR MEDICINE (Amsterdam: Elsevier B.V.) pp. - - issn: 2949-6888 - wos: (0) - scopus: (0)

11573/1642860 - 2022 - The gene promoter binding activity of miR-223 induces flotillin-1 expression and function to regulate myeloid differentiation
Billi, Monia; Ianni, Alessandro; Quattrocchi, Alberto; Gentile, Martina; Borello, Ugo; Grignani, Francesco; Nervi, Clara; De Marinis, Elisabetta - 01h Abstract in rivista
rivista: HAEMATOLOGICA (Fondaz Ferrata Storti:Strada Nuova 134, 27100 Pavia Italy:011 39 0382 531182, EMAIL: office@haematologica.it, Fax: 011 39 0382 27721) pp. 39-39 - issn: 0390-6078 - wos: WOS:000783057300068 (0) - scopus: (0)

11573/1644422 - 2022 - Daratumumab triplet therapies in patients with relapsed or refractory multiple myeloma. A “real world” experience
Fucci, Ludovica; Gensini, Lorenzo; Coppetelli, Ugo; Ortu La Barbera, Elettra; Gentile, Martina; Fiori, Luciano; Perrone, Salvatore; Cimino, Giuseppe - 01a Articolo in rivista
rivista: LEUKEMIA RESEARCH REPORTS (Amsterdam : Elsevier) pp. - - issn: 2213-0489 - wos: WOS:001051054500006 (2) - scopus: 2-s2.0-85131740324 (3)

11573/1486481 - 2020 - Genetic lesions disrupting calreticulin 3′-untranslated region in JAK2 mutation-negative polycythemia vera
Quattrocchi, A.; Maiorca, C.; Billi, M.; Tomassini, S.; De Marinis, E.; Cenfra, N.; Equitani, F.; Gentile, M.; Ceccherelli, A.; Banella, C.; Mecarocci, S.; Scerpa, M. C.; Pisano, S.; Pacilli, A.; Di Cristofano, C.; Mancini, M.; Guglielmelli, P.; Vannucchi, A. M.; Noguera, N.; Grignani, F.; Cimino, G.; Nervi, C. - 01f Lettera, Nota
rivista: AMERICAN JOURNAL OF HEMATOLOGY (Hoboken N.J.: Wiley-Blackwell) pp. 263-267 - issn: 0361-8609 - wos: WOS:000550503200001 (9) - scopus: 2-s2.0-85088144967 (9)

11573/1272651 - 2018 - HDAC inhibition by valproic acid decreases JAK2V617F levels in myeloproliferative neoplasms via up-regulation of miR-101, in vivo and in vitro
De Marinis, E.; Cenfra, N.; Liberati, D.; Scerpa, M. C.; Pagano, F.; Quattrocchi, A.; Tomassini, S.; Gentile, M.; Ceccherelli, A.; Cicalini, A.; Fantasia, F.; Cimino, G.; Nervi, C. - 01h Abstract in rivista
rivista: HEMASPHERE (New York NY: Wolters Kluwer) pp. 1-1113 - issn: 2572-9241 - wos: WOS:000531530400001 (1) - scopus: (0)

11573/1413773 - 2018 - A novel germline CALR mutation affecting an evolutionary conserved region of 3’UTR in JAK2-negative siblings with Polycythemia Vera
Quattrocchi, A.; Tomassini, S.; Scerpa, M. C.; Cenfra, N.; Pisanò, S.; De Marinis, E.; Gentile, M.; Ceccherelli, A.; Maiorca, C.; Cicalini, A.; Fantasia, F.; Cimino, G.; Nervi, C. - 01h Abstract in rivista
rivista: HEMASPHERE (New York NY: Wolters Kluwer) pp. - - issn: 2572-9241 - wos: (0) - scopus: (0)

11573/1413779 - 2018 - A novel germline mutation in the 3'Untranslated region of Calreticulin gene induces JAK/STAT signaling activation and Erythrocytosis.
Quattrocchi, Alberto; Tomassini, Simona; Billi, M.; Scerpa, Mc.; Cenfra, N.; Gentile, Martina; Maiorca, Carlo; Ceccherelli, A.; De Marinis, E.; Grignani, F.; Cimino, G.; Nervi, C. - 01h Abstract in rivista
rivista: HAEMATOLOGICA (Pavia : Fondazione Ferrata Storti) pp. - - issn: 1592-8721 - wos: WOS:000451725500079 (0) - scopus: (0)

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