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Simple Summary For the first time, neuronal features are described in blastic plasmacytoid dendritic cell neoplasm (BPDCN) by a complex array of molecular techniques, including microRNA and gene expression profiling, RNA and Chromatin immunoprecipitation sequencing, and immunohistochemistry. The discovery of unexpected neural features in BPDCN may change our vision of this disease, leading to the designing of a new BPDCN cell model and to re-thinking the relations occurring between BPDCN and nervous system. The observed findings contribute to explaining the extreme tumor aggressiveness and also to propose novel therapeutic targets. In view of this, the identification, in this work of new potential neural metastatic inducers might open the way to therapeutic approaches for BPDCN patients based on the use of anti-neurogenic agents. Abstract Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and highly aggressive hematologic malignancy originating from plasmacytoid dendritic cells (pDCs). The microRNA expression profile of BPDCN was compared to that of normal pDCs and the impact of miRNA dysregulation on the BPDCN transcriptional program was assessed. MiRNA and gene expression profiling data were integrated to obtain the BPDCN miRNA-regulatory network. The biological process mainly dysregulated by this network was predicted to be neurogenesis, a phenomenon raising growing interest in solid tumors. Neurogenesis was explored in BPDCN by querying different molecular sources (RNA sequencing, Chromatin immunoprecipitation-sequencing, and immunohistochemistry). It was shown that BPDCN cells upregulated neural mitogen genes possibly critical for tumor dissemination, expressed neuronal progenitor markers involved in cell migration, exchanged acetylcholine neurotransmitter, and overexpressed multiple neural receptors that may stimulate tumor proliferation, migration and cross-talk with the nervous system. Most neural genes upregulated in BPDCN are currently investigated as therapeutic targets.

Burkitt lymphoma (BL) is an aggressive neoplasm characterized by consistent morphology and phenotype, typical clinical behavior and distinctive molecular profile. The latter is mostly driven by the MYC over-expression associated with the characteristic translocation (8;14) (q24; q32) or with variant lesions. Additional genetic events can contribute to Burkitt Lymphoma pathobiology and retain clinical significance. A pathogenetic role for Epstein-Barr virus infection in Burkitt lymphomagenesis has been suggested; however, the exact function of the virus is largely unknown. In this study, we investigated the molecular profiles (genes and microRNAs) of Epstein-Barr virus-positive and -negative BL, to identify specific patterns relying on the differential expression and role of Epstein-Barr virus-encoded microRNAs. First, we found significant differences in the expression of viral microRNAs and in selected target genes. Among others, we identified LIN28B, CGNL1, GCET2, MRAS, PLCD4, SEL1L, SXX1, and the tyrosine kinases encoding STK10/STK33, all provided with potential pathogenetic significance. GCET2, also validated by immunohistochemistry, appeared to be a useful marker for distinguishing EBV-positive and EBV-negative cases. Further, we provided solid evidences that the EBV-encoded microRNAs (e.g. BART6) significantly mold the transcriptional landscape of Burkitt Lymphoma clones. In conclusion, our data indicated significant differences in the transcriptional profiles of EBV-positive and EBV-negative BL and highlight the role of virus encoded miRNA.

Multiple myeloma (MM) is a hematological malignancy of plasma cells in the bone marrow. Despite multiple treatment options, MM is inevitably associated with drug resistance and poor outcomes. Histone deacetylase inhibitors (HDACi's) are promising novel chemotherapeutics undergoing evaluation in clinical trials for the potential treatment of patients with MM. Although in preclinical studies HDACi's have proven anti-myeloma activity, but in the clinic single-agent HDACi treatments have been limited due to low tolerability. Improved clinical outcomes were reported only when HDACi's were combined with other drugs. Here, we show that a novel pan-HDACi AR-42 downregulates CD44, a glycoprotein that has been associated with lenalidomide and dexamethasone resistance in myeloma both in vitro and in vivo. We also show that this CD44 downregulation is in part mediated by miR-9-5p, targeting insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3), which directly binds to CD44 mRNA and increases its stability. Importantly, we also demonstrate that AR-42 enhances anti-myeloma activity of lenalidomide in primary MM cells isolated from lenalidomide resistant patients and in in vivo MM mouse model. Thus, our findings shed light on potential novel combinatorial therapeutic approaches modulating CD44 expression, which may help overcome lenalidomide resistance in myeloma patients.

hsa-mir-483 is located within intron 2 of the IGF2 locus. We found that the mature microRNA (miRNA) miR-483-3p is overexpressed in 100% of Wilms' tumors. In addition, colon, breast, and liver cancers exhibit high or even extremely high levels of miR-483-3p in ∼30% of the cases. A coregulation with IGF2 mRNA was detected, although some tumors exhibited high expression of miR-483-3p without a concomitant increase of IGF2. These findings suggested that miR-483-3p could cooperate with IGF2 or act as an autonomous oncogene. Indeed, here we prove that an anti-miRNA oligonucleotide against miR-483-3p could inhibit the miRNAs without affecting IGF2 mRNA and it could suppress tumorigenicity of HepG2 cells, a cell line that overexpresses miR-483-3p and IGF2. Conversely, no antitumor effect was elicited by inhibition of IGF2. The oncogenic mechanism of miR-483-3p was at least partially clarified by the finding that it could modulate the proapoptotic protein BBC3/PUMA and miR-483-3p enforced expression could protect cells from apoptosis. Our results indicate that miR-483-3p could function as an antiapoptotic oncogene in various human cancers and reveal a new, potentially important target for anticancer therapy. Cancer Res; 70(8); 3140–9. ©2010 AACR.

MicroRNAs (miRNAs) are noncoding single-stranded RNA molecules of ∼22 nucleotides in length. They have a critical role in regulating gene expression by targeting messenger RNAs (mRNAs) in a sequence-specific manner. Active mature miRNAs are produced from primary miRNA transcripts (pri-miRNAs) through sequential cleavages by the microprocessor complex, which includes DROSHA, DGCR8, and DICER proteins. Several miRNAs are specifically up-regulated in various types of tumors, and a wide range of studies have demonstrated how their down-regulation could potentially affect tumorigenesis, metastasis formation and drug resistance. However, most of these studies have indicated the possibility of modulating miRNA expression at the transcriptional level, while only a few focused on post-transcriptional control. Nucleolin (NCL) is a highly conserved multifunctional protein involved in ribosomal RNA (rRNA) biogenesis and in the stabilization of different mRNAs, in the nucleus and in the cytoplasm of normal and cancer cells. NCL was also found on the surface of different types of cancer cells, but not on their normal counterparts, shuttling between the inner and the outer part of the cell membrane. These observations suggested that NCL might be considered a cancer cell specific receptor, able to mediate tumor-selective uptake of specific ligands such as RNA and DNA G-rich aptamers. Here we show that NCL binds the terminal loop and promotes the maturation of a specific set of miRNAs, including miR-21, miR-103, miR-221 and miR-222, whose over-expression is causally associated with greater aggressiveness and resistance to anti-neoplastic therapies of several kind of tumors, such as breast cancer. Accordingly, a direct correlation between the expression levels of NCL and NCL-dependent miRNAs in human breast cancer samples was also observed. Conversely, NCL impairment down-regulated NCL-dependent miRNAs and up-regulates their target in vitro, affecting cancer cell proliferation, migration and anti-neoplastic drug resistance. Finally, we used AS1411, the first NCL-targeting G-rich aptamer that has reached phase II clinical trials for cancer therapy, to inhibit NCL activity in orthotopc xenograft models of breast cancer. Our in vivo data demonstrate that AS1411 down-regulates NCL-dependent miRNAs, hindering breast cancer metastasis. These findings provide insights into the molecular function of NCL in miRNA biology, as well as one of the first realistic strategies for miRNA regulation in cancer therapy. Citation Format: Dario Palmieri, Luciana De Luca, Jessica Consiglio, Alberto Rocci, Tiffany Talabere, Claudia Piovan, Alessandro Lagana, Luciano Cascione, Jingwen Guan, Pierluigi Gasparini, Veronica Balatti, Vincenzo Coppola, Craig Hofmeister, Guido Marcucci, John Byrd, Stefano Volinia, Charles Shapiro, Michael Freitas, Carlo Croce, Flavia Pichiorri. In vivo NCL-targeting affects breast cancer aggressiveness through miRNA regulation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1122. doi:10.1158/1538-7445.AM2013-1122

The hsa-mir-483 locus is located within intron 2 of the IGF2 locus. Here, we show that the mature microRNA (miRNA) miR-483-3p is over-expressed and co-regulated with IGF2 in 100% of Wilms’ tumors. Additionally, several types of common adult cancers exhibit high or even extremely high levels of miR-483-3p. We show that the mutation status of the oncoprotein β-catenin is correlated with the expression of the miR-483-3p in hepatocarcinoma and colorectal cancer. These findings suggested an autonomous oncogenic function for this miRNA. Indeed, anti-microRNA oligonucleotide (AMO) against miR-483-3p could inhibit tumorigenicity of HepG2 cells. At the same time, no anti-tumor effect was elicited by inhibition of IGF2. The oncogenic properties of miR-483-3p were further supported by the finding that its ectopic expression could protect cells from apoptosis through the regulation of one important pro-apoptotic protein, Puma. Our results indicate that miR-483-3p can function as an anti-apoptotic oncogene in various human cancers and reveal a new potentially important target for anti-cancer therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2087.