2024, Articolo in rivista, ENG
De Robertis, Mariangela; Signori, Emanuela
Recent progress in developing new vaccination strategies against cancer requires the production of complex and reliable animal models reflecting the complexity of the tumors with their microenvironment. Mice can be considered a good source due to low cost and ease of being genetically modified, inoculated with tumor cell lines or treated by chemicals to induce different cancers. Despite significant limitations in modeling human cancer complexity, preclinical trials conducted in mice can efficiently contribute to understand molecular mechanisms of cancer, to closely resemble and follow carcinogenesis steps impossible to study into humans, and to test new anticancer therapies. In this chapter, we generally describe the different mouse models developed for cancer vaccines' preclinical trials. A particular focus is dedicated to a chemically-induced colorectal cancer model in use in our laboratories.
2024, Articolo in rivista, ENG
De Robertis, Mariangela; Lampreht Tratar, Ursa; Signori, Emanuela; Komel, Tilen; Cemazar, Maja
Efficacy of novel cancer immunization protocols could be tested in cell line-derived xenograft tumor models (CDX), which are based on the implantation of human tumor cell lines into mice for the development of different tumors by numerous means, such as subcutaneous implantation and orthotopic, venial, or peritoneal injections. However, the disadvantages of this model are the biological alteration of the derived cells or the inability of the cell lines to accurately reflect the complexity of tumor heterogeneity. Therefore, syngeneic mouse models, which offer a relatively simple grafting technique, preservation of lineage hierarchy, and the ability to generate tumors in as little as 2-8 weeks, are being used to study potential future applications in medical treatment, particularly immunotherapies. Here, we describe a B16.F10 C57Bl/6 mouse melanoma model we selected for therapeutic studies employing IL-2 and IL-12 immunization protocols. Procedure of tumor cells inoculation and melanoma development in mice is described in detail, as first and necessary set-up for successful immunization experiments.
2023, Articolo in rivista, ENG
Cancelliere, Rocco; Cosio, Terenzio; Campione, Elena; Corvino, Martina; D'Amico, Maria Pia; Micheli, Laura; Signori, Emanuela; Contini, Giorgio
Interleukin-6 (IL-6) plays a crucial role in autoimmunity and chronic inflammation. This study aims to develop a low-cost, simple-to-manufacture, and user-friendly label-free electrochemical point-of-care device for the rapid detection of IL-6 in patients with psoriasis. Precisely, a sandwich-based format immunosensor was developed using two primary antibodies (mAb-IL6 clone-5 and clone-7) and screen-printed electrodes modified with an inexpensive recycling electrochemical enhancing material, called biochar. mAb-IL6 clone-5 was used as a covalently immobilized capture bioreceptor on modified electrodes, and mAb-IL6 clone-7 was used to recognize the immunocomplex (Anti-IL6 clone-5 and IL-6) and form the sandwich. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to conduct electrochemical characterization of the layer-by-layer assembly of the immunosensor, while square wave voltammetry (SWV) was used to perform the sensing. The developed immunosensor demonstrated robust analytical performance in buffer solution, with a wide linear range (LR) by varying from 2 to 250 pg/mL, a good limit of detection (LOD) of 0.78 pg/mL and reproducibility (RSD<7%). In addition, a spectrophotometric ELISA kit was employed to validate the results obtained with the label-free device by analyzing twenty-five serum samples from control and patients affected by psoriasis. A strong correlation in terms of pg/mL concentration of IL-6 was found comparing the two methods, with the advantage for our label-free biosensor of an ease use and a quicker detection time. Based on IL-6 levels, the proposed immunosensor is a dependable, non-invasive screening device capable of predicting disease onset, progression, and treatment efficacy.
2023, Articolo in rivista, ENG
Tellado, Matías; De Robertis, Mariangela; Montagna, Daniela; Giovannini, Daniela; Salgado, Sergio; Michinski, Sebastián; Signori, Emanuela; Maglietti, Felipe
Electrochemotherapy (ECT) is a standard of care in veterinary and human oncology. The treatment induces a well-characterized local immune response which is not able to induce a systemic response. In this retrospective cohort study, we evaluated the addition of gene electrotransfer (GET) of canine IL-2 peritumorally and IL-12 intramuscularly to enhance the immune response. Thirty canine patients with inoperable oral malignant melanoma were included. Ten patients received ECT+GET as the treatment group, while twenty patients received ECT as the control group. Intravenous bleomycin for the ECT was used in both groups. All patients had compromised lymph nodes which were surgically removed. Plasma levels of interleukins, local response rate, overall survival, and progression-free survival were evaluated. The results show that IL-2 and IL-12 expression peaked around days 7-14 after transfection. Both groups showed similar local response rates and overall survival times. However, progression-free survival resulted significantly better in the ECT+GET group, which is a better indicator than overall survival, as it is not influenced by the criterion used for performing euthanasia. We can conclude that the combination of ECT+GET using IL-2 and IL-12 improves treatment outcomes by slowing down tumoral progression in stage III-IV inoperable canine oral malignant melanoma.
2023, Contributo in volume, ENG
Amelia Licari 1,2,3, Giuliana Ferrante3,4, Velia Malizia 3, Agnese Augello5 and Stefania La Grutta3
Medical assistive robots (MARs) are innovative tools providing extensive support and assistance to users in different medical scenarios, enhancing patients' health and care. Social MARs have been implemented in respiratory medicine to help manage chronic respiratory conditions, such as asthma, COPD and cystic fibrosis. To integrate MARs into routine clinical practice, more studies are needed to strengthen the evidence on the feasibility, acceptability and efficacy of MARs in chronic respiratory conditions in the long term.
2023, Articolo in rivista, ENG
Beatrice Andrenacci 1, Maria De Filippo 1 2, Martina Votto 1 2, Maria Sole Prevedoni Gorone 3, Mara De Amici 4, Stefania La Grutta 5, Gian Luigi Marseglia 1 2, Amelia Licari 1 2 5
Introduction: Although rare, pediatric severe therapy-resistant asthma (STRA) is a highly heterogeneous, resource-demanding disease that differs significantly from severe adult asthma and whose pathogenesis is still poorly understood. Areas covered: This review summarizes the latest 10 years of English-written studies defining pediatric STRA endotypes using lung-specific techniques such as bronchoalveolar lavage and endobronchial biopsy. Results of the studies and limits on the field are discussed, together with some future perspectives. Expert opinion: Over the years, it has become increasingly clear that 'one size does not fit all" in asthma. However, "Does an extremely tailored size fit more than one?'. Only using multicentric, longitudinal pediatric studies, will we be able to answer. Three issues could be particularly critical for future research. First, to provide, if existing, a distinction between prepuberal STRA and puberal STRA endotypes to understand the transition from pediatric to adult STRA and to design effective, tailored therapies in adolescents, usually suffering from poorer asthma control. Second, design early treatments for pediatric airway remodeling to preserve lifelong good lung function. Finally, to better characterize inflammation before and during biological therapies, to provide clues on whether to stop or change treatments.
2023, Articolo in rivista, ENG
Daniela Fioretti +, Mario Ledda+ , Sandra Iurescia+, Raffaella Carletti , Cira Di Gioia, Maria Grazia Lolli, Rodolfo Marchese , Antonella Lisi, and Monica Rinaldi + these authors contributed equally to this work as first co-authors
Background: The regeneration of severe traumatic muscle injuries is an unsolved medical need that is relevant for civilian and military medicine. In this work, we produced a critically sized nonhealing muscle defect in a mouse model to investigate muscle degeneration/healing phases. Materials and methods: We caused a freeze injury (FI) in the biceps femoris of C57BL/6N mice. From day 1 to day 25 post-injury, we conducted histological/morphometric examinations, an analysis of the expression of genes involved in inflammation/regeneration, and an in vivo functional evaluation. Results: We found that FI activates cytosolic DNA sensing and inflammatory responses. Persistent macrophage infiltration, the prolonged expression of eMHC, the presence of centrally nucleated myofibers, and the presence of PAX7+ satellite cells at late time points and with chronic physical impairment indicated inadequate repair. By looking at stem-cell-based therapeutic protocols of muscle repair, we investigated the crosstalk between M1-biased macrophages and human amniotic mesenchymal stem cells (hAMSCs) in vitro. We demonstrated their reciprocal paracrine effects where hAMSCs induced a shift of M1 macrophages into an anti-inflammatory phenotype, and M1 macrophages promoted an increase in the expression of hAMSC immunomodulatory factors. Conclusions: Our findings support the rationale for the future use of our injury model to exploit the full potential of in vivo hAMSC transplantation following severe traumatic injuries.
2023, Altro prodotto, ITA
Daniela Fioretti; Sandra Iurescia
E-poster dal titolo "I vaccini genetici: le biotecnologie nella lotta contro il cancro". Contributo alla mostra 'Immunologia: il fascino della diversità', organizzata dal Cnr Immunology Network (CIN), fruibile dal 23 ottobre al 9 novembre, presso la sede centrale del Cnr, per celebrare il centenario del Cnr.
2023, Altro prodotto, ITA
Chiarello Fabio; Bertani Francesca Romana; Sandra Iurescia; Daniela Fioretti; Businaro Luca; Gerardino Annamaria
Laboratorio/ evento CNR al Festival della Scienza di Genova 2023; diffusione della cultura scientifica In collaborazione con CNR - Unità Comunicazione e Rome Technopole; - "Impronte spettrali" è un laboratorio basato su giochi ed attività ludiche, pensato in particolare per bambini e ragazzi delle scuole elementari e medie ma adatto a tutti. Lo scopo è illustrare in modo pratico i concetti e le applicazioni della spettroscopia, raccontando attraverso il gioco e la sperimentazione diretta come sia possibile utilizzare lo studio dello spettro luminoso per trovare "impronte", tracce che ci permettono di ricavare informazioni nascoste per applicazioni in campi diversi come l'astronomia, la chimica, la biologia, la fisica, la scienza dei materiali, l'agrifood, le scienze forensi.
2023, Altro prodotto, ITA
Daniela Fioretti; Sandra Iurescia
l linguaggio della Vita - Viaggio fantastico dal DNA alle proteine: laboratorio basato su giochi e attività ludiche, pensato in particolare per bambini e ragazzi delle scuole elementari e medie ma adatto a tutti. Lo scopo è illustrare in modo pratico il "linguaggio della vita", raccontando attraverso il gioco come funziona la riproduzione cellulare, dal DNA all'RNA fino agli amminoacidi e alla produzione delle proteine. (Padiglione LEARN)
2023, Articolo in rivista, ENG
S. Bonomo 1*, P. Marchetti 2, S. Fasola 3, R. Vesentini 2, A. Marcon 2, G. Ferrante 4, L. Antonicelli 5, S. Battaglia 6, R. Bono 7, G. Squillacioti 7, N. Murgia 8, P. Pirina 9, S. Villani 10, S. La Grutta 3, G. Verlato 2 & G. Viegi 11
An association between climatic conditions and asthma incidence has been widely assumed. However, it is unclear whether climatic variations have a fingerprint on asthma dynamics over long time intervals. The aim of this study is to detect a possible correlation of the Summer North Atlantic Oscillation (S-NAO) index and the self-calibrated palmer drought severity index (scPDSI) with asthma incidence over the period from 1957 to 2006 in Italy. To this aim, an analysis of non-stationary and non-linear signals was performed on the time series of the Italian databases on respiratory health (ISAYA and GEIRD) including 36,255 individuals overall, S-NAO, and scPDSI indices to search for characteristic periodicities. The ISAYA (Italian Study on Asthma in Young Adults) and GEIRD (Gene Environment Interactions in Respiratory Diseases) studies collected information on respiratory health in general population samples, born between 1925 and 1989 and aged 20-84 years at the time of the interview, from 13 Italian centres. We found that annual asthma total incidence shared the same periodicity throughout the 1957-2006 time interval. Asthma incidence turned out to be correlated with the dynamics of the scPDSI, modulated by the S-NAO, sharing the same averaged 6 year-periodicity. Since climate patterns appear to influence asthma incidence, future studies aimed at elucidating the complex relationships between climate and asthma incidence are warranted.
2023, Articolo in rivista, ENG
Federico Mei1,2*+, Matteo Renzi3*+, Martina Bonifazi1,2, Floriano Bonifazi4, Nicola Pepe5, Alessio D'Allura5, Giuseppe Brusasca5, Giovanni Viegi6 and Francesco Forastiere7,8
Background Shipping and port-related air pollution has a significant health impact on a global scale. The present study aimed to assess the mortality burden attributable to long-term exposure to ambient particulate matter (PM2.5, PM10) and nitrogen dioxide (NO2) in the city of Ancona (Italy), with one of the leading national commercial harbours. Methods Exposure to air pollutants was derived by dispersion models. The relationship between the long-term exposure of air pollution exposure and cause-specific mortality was evaluated by Poisson regression models, after adjustment for gender, age and socioeconomic status. Results are expressed as percent change of risk (and relative 95% confidence intervals) per 5 unit increases in the exposures. The health impact on the annual number of premature cause-specific deaths was also assessed. Results PM2.5 and NO2 annual concentrations were higher in the area close to the harbour than in the rest of the city. Positive associations between each pollutant and most of the mortality outcomes were observed, with estimates of up to 7.6% (95%CI 0.1, 15.6%) for 10 ?g/m3 increase in NO2 and cardiovascular mortality and 15.3% (95%CI-1.1, 37.2%) for 10 ?g/m3 increase PM2.5 and lung cancer. In the subpopulation living close to the harbour, there were excess risks of up to 13.5%, 24.1% and 37.9% for natural, cardiovascular and respiratory mortality. The number of annual premature deaths due to the excess of PM2.5 and NO2 exposure (having as a reference the 2021 World Health Organization Air Quality Guidelines) was 82 and 25, respectively. Conclusions Our study confirms the long-term health effects of PM and NO2 on mortality and reveals a higher mortality burden in areas close to shipping and port-related emissions. Estimating the source-specific health burdens is key to achieve a deeper understanding of the role of different emission sources, as well as to support effective and targeted mitigation strategies.
2023, Articolo in rivista, ENG
Joerg S. Steier, Richard K. Bogan, Irene M. Cano-Pumarega , John A. Fleetham, Giuseppe Insalaco, Chitra Lal, Jean-Louis Pepin, Winfried J. Randerath, Susan Redline, Atul Malhotra
Study objective: Excessive daytime sleepiness is common with obstructive sleep apnoea and can persist despite efforts to optimise primary airway therapy. The literature lacks recommendations regarding differential diagnosis and management of excessive daytime sleepiness in obstructive sleep apnoea. This study sought to develop expert consensus statements to bridge the gap between existing literature/guidelines and clinical practice. Methods: A panel of 10 international experts was convened to undertake a modified Delphi process. Statements were developed based on available evidence identified through a scoping literature review, and expert opinion. Consensus was achieved through 3 rounds of iterative, blinded survey voting and revision to statements until a predetermined level of agreement was met (>=80 % voting "strongly agree" or "agree with reservation"). Results: Consensus was achieved for 32 final statements. The panel agreed excessive daytime sleepiness is a patient-reported symptom. The importance of subjective/objective evaluation of excessive daytime sleepiness in the initial evaluation and serial management of obstructive sleep apnoea was recognised. The differential diagnosis of residual excessive daytime sleepiness in obstructive sleep apnoea was discussed. Optimizing airway therapy (eg, troubleshooting issues affecting effectiveness) was addressed. The panel recognised occurrence of residual excessive daytime sleepiness in obstructive sleep apnoea despite optimal airway therapy and the need to evaluate patients for underlying causes. Conclusions: Excessive daytime sleepiness in patients with obstructive sleep apnoea is a public health issue requiring increased awareness, recognition, and attention. Implementation of these statements may improve patient care, long-term management, and clinical outcomes in patients with obstructive sleep apnoea.
2023, Contributo in atti di convegno, ENG
Fabio Chiarello; Daniela Fioretti; Tiziano Antognozzi; Francesca Romana Bertani; Ennio Bilancini; Matteo Bisanti; Leonardo Boncinelli; Luca Businaro; Roberto Di Paolo; Annamaria Gerardino; Sandra Iurescia
Protein biosynthesis is a complex process that involves the transcription of DNA into mRNA and the subsequent translation of mRNA into proteins according to the genetic code. To introduce this fundamental process to a broad audience, we developed "The Language of Life", a game-based workshop that was presented at the Genoa Science Festival 2022, the largest science communication event in Italy. The game-based workshop employed jigsaw puzzle-like elements to represent DNA, mRNA, and aminoacyl transfer RNAs, enabling participants to pair them through codon combinations. The game-based workshop lasted for an hour and the framing was a "special mission" inside a cell. It consisted of an initial training phase that incorporated videos, models, and explanations, followed by practical "missions'' in which participants reproduce transcription and translation mechanisms by moving inside the cell and using the provided materials. The workshop was attended by 1,505 participants, primarily students aged 6-18, and received positive feedback. In this paper, we present our experience conducting this workshop and discuss its impact and potential for future use.
2023, Altro prodotto, ITA
Daniela Fioretti; Sandra Iurescia
Evento divulgativo dell'IFT-CNR tenutosi presso l'Area di Ricerca Roma2 Tor Vergata il 28 settembre 2023, nell'ambito della European Biotech Week (EBW) 2023, manifestazione divulgativa mondiale organizzata in Italia da Associazione Nazionale per lo sviluppo delle Biotecnologie (Assobiotec )- Federchimica e da EuropaBio (Associazione europea delle biotecnologie). L' evento, dedicato a "Vaccini genetici: le biotecnologie nella lotta contro il cancro", racconta come è possibile "educare" il nostro sistema immunitario e usarlo come alleato nella lotta contro i tumori. L'appuntamento prevede un'attività dedicata agli studenti della scuola secondaria superiore: un laboratorio scientifico-formativo che, attraverso metodologie interattive, intende trasferire le conoscenze del settore biotecnologico nel campo dei vaccini. I partecipanti verranno accompagnati in un percorso didattico dove potranno sperimentare la "realizzazione" di un vaccino genetico anti-tumorale con l'uso di modelli 2-D di DNA e RNA.
2023, Altro prodotto, ENG
Angelica Canossi1 · Alessandra Panarese2 · Valentina Savino2 · Tiziana Del Beato1 · Francesco Pisani2
In this study we describe the impact of COVID-19 infection on the immunological response of kidney transplant recipients of a single transplant center. We evaluated the seroconversion after vaccination and COVID-19 disease, the management of immunosuppressive therapy, and the effects of the withdrawal of immunosuppressive therapy on renal function and clinical outcome.
2023, Articolo in rivista, CPE
Antonio Gatto, Lavinia Capossela, Giorgio Conti, Gemma Eftimiadi, Serena Ferretti, Luigi Manni, Antonietta Curatola, Benedetta Graglia, Lorenzo Di Sarno, Maria Lucia Calcagni, Daniela Di Giuda, Stefano Cecere, Domenico Marco Romeo, Marzia Soligo, Enzo Picconi, Marco Piastra, Giacomo Della Marca, Susanna Staccioli, Antonio Ruggiero, Fabrizio Cocciolillo, Silvia Pulitanò & Antonio Chiaretti
Background Severe traumatic brain injury (TBI) is one of the most dramatic events in pediatric age and, despite advanced neuro-intensive care, the survival rate of these patients remains low. Children suffering from severe TBI show long-term sequelae, more pronounced in behavioral, neurological and neuropsychological functions leading to, in the most severe cases, an unresponsive wakefulness syndrome (UWS). Currently, no effective treatments can restore neuronal loss or produce significant improvement in these patients. In experimental animal models, human- recombinant Nerve Growth Factor (hr-NGF) promotes neural recovery supporting neuronal growth, differentiation and survival of brain cells and up-regulating the neurogenesis-associated processes. Only a few studies reported the efficacy of intranasal hr-NGF administration in children with post- traumatic UWS. Methods Children with the diagnosis of post-traumatic UWS were enrolled. These patients underwent a treatment with intranasal hr-NGF administration, at a total dose of 50 gamma/kg, three times a day for 7 consecutive days. The treatment schedule was performed for 4 cycles, at one month distance each. Neuroradiogical evaluation by Positron Emission Tomography scan (PET), Single Photon Emission Computed Tomography (SPECT), Electroencephalography (EEG), and Power Spectral Density (PSD) was determined before the treatment and one month after the end. Neurological assessment was also deepened by using modified Ashworth Scale, Gross Motor Function Measure, and Disability Rating Scale. Results Three children with post-traumatic UWS were treated. hr-NGF administration improved functional (PET and SPECT) and electrophysiological (EEG and PSD) assessment. Also clinical conditions improved, mainly for the reduction of spasticity and with the acquisition of voluntary movements, facial mimicry, attention and verbal comprehension, ability to cry, cough reflex, oral motility, and feeding capacity, with a significant improvement of their neurological scores. No side effects were reported. Conclusion These promising results and the ease of administration of this treatment make it worthwhile to be investigated further, mainly in the early stages from severe TBI and in patients with better baseline neurological conditions, to explore more thoroughly the benefits of this new approach on neuronal function recovery after traumatic brain damage.
2023, Articolo in rivista, ENG
Verdile V, Palombo R, Ferrante G, Ferri A, Amadio S, Volonté C, Paronetto MP
Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disease characterized by the degeneration of upper and lower motor neurons, progressive wasting and paralysis of voluntary muscles. A hallmark of ALS is the frequent nuclear loss and cytoplasmic accumulation of RNA binding proteins (RBPs) in motor neurons (MN), which leads to aberrant alternative splicing regulation. However, whether altered splicing patterns are also present in familial models of ALS without mutations in RBP-encoding genes has not been investigated yet. Herein, we found that altered splicing of synaptic genes is a common trait of familial ALS MNs. Similar deregulation was also observed in hSOD1G93A MN-like cells. In silico analysis identified the potential regulators of these pre-mRNAs, including the RBP Sam68. Immunofluorescence analysis and biochemical fractionation experiments revealed that Sam68 accumulates in the cytoplasmic insoluble ribonucleoprotein fraction of MN. Remarkably, the synaptic splicing events deregulated in ALS MNs were also affected in Sam68-/- spinal cords. Recombinant expression of Sam68 protein was sufficient to rescue these splicing changes in ALS hSOD1G93A MN-like cells. Hence, our study highlights an aberrant function of Sam68, which leads to splicing changes in synaptic genes and may contribute to the MN phenotype that characterizes ALS.
2023, Articolo in rivista, CPE
Sara Piacentini 1, Anna Riccio 1, Silvia Santopolo 1, Silvia Pauciullo 1, Simone La Frazia 1, Antonio Rossi 2, Jean-Francois Rossignol 3, M Gabriella Santoro 1 2
Coronaviridae is recognized as one of the most rapidly evolving virus family as a consequence of the high genomic nucleotide substitution rates and recombination. The family comprises a large number of enveloped, positive-sense single-stranded RNA viruses, causing an array of diseases of varying severity in animals and humans. To date, seven human coronaviruses (HCoV) have been identified, namely HCoV-229E, HCoV-NL63, HCoV-OC43 and HCoV-HKU1, which are globally circulating in the human population (seasonal HCoV, sHCoV), and the highly pathogenic SARS-CoV, MERS-CoV and SARS-CoV-2. Seasonal HCoV are estimated to contribute to 15-30% of common cold cases in humans; although diseases are generally self-limiting, sHCoV can sometimes cause severe lower respiratory infections and life-threatening diseases in a subset of patients. No specific treatment is presently available for sHCoV infections. Herein we show that the anti-infective drug nitazoxanide has a potent antiviral activity against three human endemic coronaviruses, the Alpha-coronaviruses HCoV-229E and HCoV-NL63, and the Beta-coronavirus HCoV-OC43 in cell culture with IC50 ranging between 0.05 and 0.15 ?g/mL and high selectivity indexes. We found that nitazoxanide does not affect HCoV adsorption, entry or uncoating, but acts at postentry level and interferes with the spike glycoprotein maturation, hampering its terminal glycosylation at an endoglycosidase H-sensitive stage. Altogether the results indicate that nitazoxanide, due to its broad-spectrum anti-coronavirus activity, may represent a readily available useful tool in the treatment of seasonal coronavirus infections.