Ribosome biogenesis rate, a parameter of sensitivity to chemotherapeutic drugs inhibiting rRNA synthesis Davide Treré, Lorenzo Montanaro, Massimo Derenzini, Claudio Agostinelli, Enrico Derenzini Oncology Reviews, 2026 Many drugs currently used in cancer chemotherapy exert their toxic action mainly by inhibiting ribosome biogenesis (RiBi). This is due to the fact that after inhibition of rRNA transcription ribosomal proteins, no longer used for ribosome building, bind to and neutralize the activity of the murine double minute 2 protein (MDM2, HMD2 in humans), thus hindering cell proliferation and possibly inducing apoptotic cell death. Here, we discuss the existing literature showing how RiBi rate and genomic alterations of ribosomal proteins (RP mutations/deletions) influence the degree of MDM2 inhibition after treatment with RiBi inhibitors in cancer cells. There is evidence that a high RiBi rate is associated with a high RPs release with strong inhibition of MDM2 activity and consequent induction of apoptotic cell death in response to RiBi inhibitors, whereas a low RiBi rate or RP mutations/deletions are associated with a degree of MDM2 inhibition insufficient to kill cancer cells. In the latter case, in cells with wild type p53, association with drugs which stabilize p53 with different mechanisms may overcome cancer cells resistance to RiBi inhibition, whereas in cancers lacking functional p53 addition of MDM2 inhibitors should be considered. From this, the necessity to evaluate the rate of ribosome biogenesis together with the presence of RP mutations/deletions in cancer tissues for predicting the sensitivity of cancer cells to RiBi inhibitors in order to choose more appropriate therapeutic protocols.
5.8S rRNA forms and ribosome heterogeneity in breast cancer Giulia Venturi, Federico Zacchini, Francesca Ruzzi, Angelo Gianluca Corradini, Margherita Serra, Marianna Penzo, Davide Treré, Pier-Luigi Lollini, Lorenzo Montanaro Biochimie, 2025 Ribosome heterogeneity can contribute to translation regulation in terms of mRNA selection and translation efficiency. This is particularly true for cancer cells in which oncoribosomes are reported to translate mRNAs encoding for proteins involved in cancer progression. Among other factors, a source of ribosome heterogeneity not yet characterized could be represented by 5.8S rRNA and its three forms distinguished based on their 5' sequence. So far, little is known about the role of the presence of these isoforms in mature ribosomes and how they may contribute to human pathology. Here we investigated the relative abundance of the three 5.8S rRNA isoforms in different contexts. Analyzing total and polysomal RNA from cancer cell lines we detected all the three forms recruited in actively translating ribosomes consistently to the basal levels of their expression. Moreover, we showed that changes in the relative abundance of 5.8S rRNA isoforms can be linked to the process of tumorigenesis in a human HER2 transgenic mouse model which develops spontaneous mammary carcinomas. Finally, from the analysis of breast cancer samples, we observed significant correlations between tumor grade, estrogen receptor status and patient prognosis with the relative abundance of 5.8S rRNA isoforms. These results suggest an additional level of complexity involving 5.8S rRNA and ribosome heterogeneity in cancer pathology.
Characterization of small nucleolar RNA retaining transcripts in human normal and cancer cells Guglielmo Rambaldelli, Sidra Asghar, Giulia Venturi, Federico Zacchini, Margherita Serra, Catia Giovannini, Laura Gramantieri, Marco Bernini, Alberto Inga, Erik Dassi, Lorenzo Montanaro Non Coding RNA Research, 2025 Small nucleolar RNAs are non-coding RNAs typically encoded within the introns of both protein-coding and non-coding genes. Interestingly, a significant fraction of snoRNA sequences is found as retained introns of specific mRNA isoforms expressed from their host gene. In the present study, we aimed to define the representation of small nucleolar RNA retaining transcripts across various human cell types and tissues including cancer. We found that these type of transcripts are widely represented in normal tissues and cancer-derived cell lines, appearing both in their full-length form and, frequently, in a shorter variant. We characterized the shortening position, which occurs at or very close to the retained small nucleolar RNA sequence at the 5' end. Interestingly, for some transcripts this shorter variant represents the only form detected. In addition, some of the small nucleolar RNA retaining transcripts can be localized into the cellular cytoplasmic fraction. Moreover, our findings point out that a variable but consistent proportion of small nucleolar RNA sequences in cells, tissues, and liquid biopsy samples is, in fact, present as small nucleolar RNA retaining transcripts, indicating that these elements should be carefully considered when snoRNA are evaluated as biomarkers. Considering that short reads and gene-based transcriptomic analysis completely overlooked these transcripts, potentially missing critical insights into their involvement in cancer and other diseases, our results strongly indicate that these type of transcripts should be further investigated in different contexts to better understand their biogenesis, sequence features, presence, and role within cells.
STK11 mutations correlate with poor prognosis for advanced NSCLC treated with first-line immunotherapy or chemo-immunotherapy according to KRAS, TP53, KEAP1, and SMARCA4 status Andrea De Giglio, Dario De Biase, Valentina Favorito, Thais Maloberti, Alessandro Di Federico, Federico Zacchini, Giulia Venturi, Claudia Parisi, Filippo Gustavo Dall’Olio, Ilaria Ricciotti, Ambrogio Gagliano, Barbara Melotti, Francesca Sperandi, Annalisa Altimari, Elisa Gruppioni, Giovanni Tallini, Francesco Gelsomino, Lorenzo Montanaro, Andrea Ardizzoni Lung Cancer, 2025 BACKGROUND: The upfront treatment of non-oncogene-addicted NSCLC relies on immunotherapy alone (ICI) or in combination with chemotherapy (CT-ICI). Genomic aberrations such as KRAS, TP53, KEAP1, SMARCA4, or STK11 may impact survival outcomes. METHODS: We performed an observational study of 145 patients treated with first-line IO or CT-ICI for advanced non-squamous (nsq) NSCLC at our institution tested with an extensive lab-developed NGS panel. The primary objective was to assess the clinical outcomes of STK11-mutated patients. Then, we performed an external validation through the public OAK/POPLAR dataset, including nsq NSCLC patients treated with single-agent ICI or CT. RESULTS: Most patients were male (59.7 %), former smokers (61.1 %), with ECOG PS 0-1 (84 %), and received first-line CT-IO (58.6 %). 44.8 % had a mutation in KRAS, 21.4 % in KEAP1, 50.3 % in TP53, 13.1 % in SMARCA4, and 14.4 % in the STK11 gene. The mOS was 8 mo. (95 % CI, 5-16.7) for STK11 mutated pts and 17.3 mo. for STK11 wild-type patients (95 % CI, 8.9-24.4) (p = 0.038). TP53 (8.3 vs 17.3), KRAS (9.2 vs 15.9), and KEAP1 (8.9 vs 15.9) mutated patients evidenced a trend for dismal mOS. SMARCA4 status had no impact on mOS. STK11 mutations were detrimental to OS in the univariate (HR 1.74, p = 0.041) and multivariate model (HR 1.97, p = 0.025) after adjusting for sex, age, ECOG PS, treatment (ICI vs CT-ICI), KRAS, KEAP1, TP53, and SMARCA4 status. Genomic alterations did not impact the mPFS in our cohort. Within the OAK/POPLAR dataset, STK11 mutations (60/818 pts) were significantly associated with increased death risk in the univariate (HR 2.01, p < 0.001) and multivariate model (HR 1.66, p = 0.001) after adjusting for age, sex, treatment (ICI vs CT), KRAS, KEAP1, TP53, and SMARCA4 status. CONCLUSION: STK11 aberrations hampered the mOS of nsq NSCLC patients treated with first-line ICI or CT-ICI. The negative prognostic impact seems to be unrelated to ICI administration.
Early monitoring of plasma KRAS G12C with digital PCR predicts antitumor response to immunotherapy or sotorasib in advanced NSCLC: A brief report Andrea De Giglio, Federico Zacchini, Giulia Venturi, Alessandro Di Federico, Claudia Parisi, Filippo Gustavo Dall’Olio, Ilaria Ricciotti, Valentina Favorito, Ambrogio Gagliano, Dario De Biase, Thais Maloberti, Annalisa Altimari, Elisa Gruppioni, Giovanni Tallini, Barbara Melotti, Francesca Sperandi, Francesco Gelsomino, Lorenzo Montanaro, Andrea Ardizzoni Journal of Liquid Biopsy, 2024 Background: The frontline management of non-oncogene addicted non-small cell lung cancer (NSCLC) involves immune-checkpoint inhibitors (ICI) alone or combined with chemotherapy (CT-ICI). The dynamic landscape of KRAS-positive NSCLC presents a spectrum of treatment options, including ICIs, targeted therapy and combination strategies currently under investigation. Methods: We conducted a prospective project to detect circulating tumor DNA (ctDNA) in patients with KRAS G12C, advanced NSCLC. We included patients undergoing upfront ICIs or subsequent line sotorasib. We planned three-time points: baseline (T0), after 3 months of treatment (T1) and at disease progression (T2). Results: 24 consecutive patients have been included. The most frequent baseline characteristics were: nonsquamous histology (95.8%), male gender (62.5%), ECOG PS 0-1 (79.2%), <3 metastatic sites (13/24, 54.2%). 18 patients (75%) received ICI-based strategies and 6 patients (25%) sotorasib. Patients with liver metastases (p = 0.01) and those with >3 metastatic sites (p = 0.002) exhibited significantly elevated ctDNA. Median overall survival (OS) was 7.5 months, progression-free survival (PFS) was 4.0 months and the objective response rate (ORR) was 33.3%. Higher AF correlated with an increased risk of death (HR 1.04, p = 0.03), though not progression. The mOS was 7.5 months (95% CI, 1.91-NR) in high-AF group and 11.3 months (95% CI, 6.6-NR) in low-AF group (p = 0.38). Notably, a reduction in plasma DNA levels was significantly associated with objective response (p = 0.01). Two patients received a T2 dosage showing increased ctDNA levels after a previous reduction associated with response. Conclusion: Early monitoring with ctDNA may offer potential benefits in the evolving scenario of KRAS G12C NSCLC treatment.
How snoRNAs can contribute to cancer at multiple levels Federico Zacchini, Chiara Barozzi, Giulia Venturi, Lorenzo Montanaro Nar Cancer, 2024 snoRNAs are a class of non-coding RNAs known to guide site specifically RNA modifications such as 2′-O-methylation and pseudouridylation. Recent results regarding snoRNA alterations in cancer has been made available and suggest their potential evaluation as diagnostic and prognostic biomarkers. A large part of these data, however, was not consistently confirmed and failed to provide mechanistic insights on the contribution of altered snoRNA expression to the neoplastic process. Here, we aim to critically review the available literature on snoRNA in cancer focusing on the studies elucidating the functional consequences of their deregulation. Beyond the canonical guide function in RNA processing and modification we also considered additional roles in which snoRNA, in various forms and through different modalities, are involved and that have been recently reported.
Decoding Ribosome Heterogeneity: A New Horizon in Cancer Therapy Valerio Gelfo, Giulia Venturi, Federico Zacchini, Lorenzo Montanaro Biomedicines, 2024 The traditional perception of ribosomes as uniform molecular machines has been revolutionized by recent discoveries, revealing a complex landscape of ribosomal heterogeneity. Opposing the conventional belief in interchangeable ribosomal entities, emerging studies underscore the existence of specialized ribosomes, each possessing unique compositions and functions. Factors such as cellular and tissue specificity, developmental and physiological states, and external stimuli, including circadian rhythms, significantly influence ribosome compositions. For instance, muscle cells and neurons are characterized by distinct ribosomal protein sets and dynamic behaviors, respectively. Furthermore, alternative forms of ribosomal RNA (rRNAs) and their post-transcriptional modifications add another dimension to this heterogeneity. These variations, orchestrated by spatial, temporal, and conditional factors, enable the manifestation of a broad spectrum of specialized ribosomes, each tailored for potentially distinct functions. Such specialization not only impacts mRNA translation and gene expression but also holds significant implications for broader biological contexts, notably in the realm of cancer research. As the understanding of ribosomal diversity deepens, it also paves the way for exploring novel avenues in cellular function and offers a fresh perspective on the molecular intricacies of translation.
Analysis of EVs from patients with advanced pancreatic cancer identifies antigens and miRNAs with predictive value Ivan Vannini, Tania Rossi, Mattia Melloni, Martina Valgiusti, Milena Urbini, Alessandro Passardi, Giulia Bartolini, Chiara Gallio, Irene Azzali, Sara Bandini, Valentina Ancarani, Lorenzo Montanaro, Giovanni Luca Frassineti, Francesco Fabbri, Ilario Giovanni Rapposelli Molecular Therapy Methods and Clinical Development, 2023 The identification of predictive factors for treatment of pancreatic cancer (PC) is an unmet clinical need. In the present work, we analyzed blood-derived extracellular vesicles (EVs) from patients with advanced PC in order to find a molecular signature predictive of response to therapy. We analyzed samples from 21 patients with advanced PC, all receiving first-line treatment with gemcitabine + nab-paclitaxel. Isolated EVs have been analyzed, and the results of laboratory have been matched with clinical data in order to investigate possible predictive factors. EV concentration and size were similar between responder and non-responder patients. Analysis of 37 EV surface epitopes showed a decreased expression of SSEA4 and CD81 in responder patients. We detected more than 450 expressed miRNAs in EVs. A comparative survey between responder and non-responder patients showed that at least 44 miRNAs were differently expressed. Some of these miRNAs have already been observed in relation to the survival and gemcitabine sensitivity of tumor cells. In conclusion, we showed the ability of our approach to identify EV-derived biomarkers with predictive value for therapy response in PC. Our findings are worthy of further investigation, including the analysis of samples from patients treated with different schedules and in different settings.
Current practice in bicistronic ires reporter use: A systematic review Guus Gijsbertus Hubert van den Akker, Federico Zacchini, Bas Adrianus Catharina Housmans, Laura van der Vloet, Marjolein Maria Johanna Caron, Lorenzo Montanaro, Tim Johannes Maria Welting International Journal of Molecular Sciences, 2021
Ribosomal protein gene RPL9 variants can differentially impair ribosome function and cellular metabolism Marco Lezzerini, Marianna Penzo, Marie-Françoise O’Donohue, Carolina Marques dos Santos Vieira, Manon Saby, Hyung L Elfrink, Illja J Diets, Anne-Marie Hesse, Yohann Couté, Marc Gastou, Alexandra Nin-Velez, Peter G J Nikkels, Alexandra N Olson, Evelien Zonneveld-Huijssoon, Marjolijn C J Jongmans, GuangJun Zhang, Michel van Weeghel, Riekelt H Houtkooper, Marcin W Wlodarski, Roland P Kuiper, Marc B Bierings, Jutte van der Werff ten Bosch, Thierry Leblanc, Lorenzo Montanaro, Jonathan D Dinman, Lydie Da Costa, Pierre-Emmanuel Gleizes, Alyson W MacInnes Nucleic Acids Research, 2020
Loss of Proteostasis Is a Pathomechanism in Cockayne Syndrome Marius Costel Alupei, Pallab Maity, Philipp Ralf Esser, Ioanna Krikki, Francesca Tuorto, Rosanna Parlato, Marianna Penzo, Adrian Schelling, Vincent Laugel, Lorenzo Montanaro, Karin Scharffetter-Kochanek, Sebastian Iben Cell Reports, 2018
Controversial relationship between the expression of the RB pathway components and RB protein phosphorylation in human breast cancer Histology and Histopathology, 2007
Relationship between the RB1 mRNA level and the expression of phosphorylated RB protein in human breast cancers: Their relevance in cell proliferation activity and patient clinical outcome Histology and Histopathology, 2007
Detection of EWS chimeric transcripts by nested RT-PCR to allow reinfusion of uncontaminated peripheral blood stem cells in high-risk Ewing's tumor in childhood Haematologica, 1999
Detection of fibronectin-binding protein genes in staphylococcal strains from peri-prosthesis infections New Microbiologica, 1999
Nucleolar function and size in cancer cells American Journal of Pathology, 1998
Primary endocervical extraosseous Ewing's sarcoma/PNET Giovanna Cenacchi, Gianandrea Pasquinelli, Lorenzo Montanaro, Serenella Cerasoli, Manuela Vici, Michele Bisceglia, Felice Giangaspero, Giuseppe N. Martinelli, Massimo Derenzini International Journal of Gynecological Pathology, 1998
N-myc amplification and cell proliferation rate in human neuroblastoma. European Journal of Histochemistry Ejh, 1997
Relationship between quantitative expression of protein p120 and proliferative activity in cancer cells. European Journal of Histochemistry Ejh, 1997
AgNOR protein expression and tumor growth rate of human carcinoma xenografts growing subcutaneously in nude mice. European Journal of Histochemistry Ejh, 1997
Cytofluorimetric and functional analysis of c-kit receptor in acute leukemia Francesco Lauria, Gian Paolo Bagnara, Damiano Rondelli, Donatella Raspadori, Pierluigi Strippoli, Laura Bonsi, Maria Alessandra Ventura, Lorenzo L. Montanaro, Giuliana Bubola, Sante Tura, Virginia C. Broudy Leukemia and Lymphoma, 1995
