Efficacy and safety of long-term treatment with budesonide orodispersible tablets in patients with Eosinophilic Esophagitis Gaia Pellegatta, Francesca Paola Giugliano, Valeria Poletti, Giacomo Marcozzi, Emanuela Morenghi, et al. Clinical and Translational Gastroenterology, 2026 Introduction and aims: Orodispersible budesonide tablets (BOT) has improved eosinophilic esophagitis (EoE) outcomes. This study aims to evaluate in real-life the efficacy of BOT and adverse events in induction and maintenance therapy. Methods: EoE patients receiving BOT (2 mg/day induction, 1 mg/day maintenance) were retrospectively analyzed over a 1-year follow-up. Clinical (SDI), endoscopic (EREFS), and histological (eosinophils<15/HPF) responses were assessed at 12 weeks (T1) and 52 weeks (T2). Results: Among 104 patients enrolled, at T1 97.8% of patients achieved histologic remission and 70.3% maintained remission at T2 with a significant improvement of EREFS and SDI. Higher SDI during induction (OR:0.72; 95%CI: 0.56-0.93) and maintenance (OR:0.65; 95%CI: 0.52-0.82) predicted loss of histologic remission at T2. BOT-PPI co-administration during maintenance therapy increased the probability for histological response at T2 (OR: 7.8; 95%CI:2.4-25.4). After induction, significant increase in incidence of erosive esophagitis (EE) was observed (2.2% vs 13.0%, p=0.013), with BOT-PPI co-administration being protective towards it (OR: 0.14; 95%CI:0.03-0.54). Local candidiasis was the most common AE at T1 and T2. Risk factors for candidiasis included a history of GERD (OR:3.71; 95%CI:1.14-12.09), EoE family history (OR: 16.94; 95% CI: 1.78-161.4), oral allergy syndrome (OR:3.53; 95%CI:1.29-9.66). Conclusion: BOT is effective and overall safe in inducing and maintaining EoE remission. EE incidence is higher during BOT therapy, supporting BOT-PPI combination in select cases. Clinical monitoring with SDI may help guide maintenance therapy and follow-up timing.
Identifying colorectal cancer-specific vulnerabilities in the Wnt-driven long non-coding transcriptome Laura J Schwarzmueller, Ronja S Adam, Leandro F Moreno, Lisanne E Nijman, Adrian Logiantara, et al. Gut, 2025 BackgroundAberrant Wnt pathway activation is a key driver of colorectal cancer (CRC) and is essential to sustain tumour growth and progression. Although the downstream protein-coding target genes of the Wnt cascade are well known, the long non-coding transcriptome has not yet been fully resolved.ObjectiveIn this study, we aim to comprehensively reveal the Wnt-regulated long non-coding transcriptome and exploit essential molecules as novel therapeutic targets.DesignWe used global run-on sequencing to define β-catenin-regulated long non-coding RNAs (lncRNAs) in CRC. CRISPRi dropout screens were subsequently used to establish the functional relevance of a subset of these lncRNAs for long-term expansion of CRC.ResultsWe uncovered thatLINC02418is essential for cancer cell clonogenic outgrowth. Mechanistically,LINC02418regulates MYC expression levels to promote CRC stem cell functionality and prevent terminal differentiation. Furthermore, we developed effective small interfering RNA (siRNA)-based therapeutics to targetLINC02418RNAin vivo.ConclusionWe propose that cancer-specific Wnt-regulated lncRNAs provide novel therapeutic opportunities to interfere with the Wnt pathway, which has so far defied effective pharmacological inhibition.
Monitoring Intestinal Organoid-Derived Monolayer Barrier Functions with Electric Cell-Substrate Impedance Sensing (ECIS) Sarah Ouahoud, Francesca Giugliano, Vanesa Muncan Bio Protocol, 2024 The measurement of transepithelial electrical resistance across confluent cell monolayer systems is the most commonly used technique to study intestinal barrier development and integrity. Electric cell substrate impedance sensing (ECIS) is a real-time, label-free, impedance-based method used to study various cell behaviors such as cell growth, viability, migration, and barrier function in vitro. So far, the ECIS technology has exclusively been performed on cell lines. Organoids, however, are cultured from tissue-specific stem cells, which better recapitulate cell functions and the heterogeneity of the parent tissue than cell lines and are therefore more physiologically relevant for research and modeling of human diseases. In this protocol paper, we demonstrate that ECIS technology can be successfully applied on 2D monolayers generated from patient-derived intestinal organoids. Key features • We present a protocol that allows the assessment of various cell functions, such as proliferation and barrier formation, with ECIS on organoid-derived monolayers. • The protocol facilitates intestinal barrier research on patient tissue-derived organoids, providing a valuable tool for disease modeling.
Amino acid variation at VP1-145 of enterovirus A71 determines the viral infectivity and receptor usage in a primary human intestinal model Ikrame Aknouch, Inés García-Rodríguez, Francesca Paola Giugliano, Carlemi Calitz, Gerrit Koen, et al. Frontiers in Microbiology, 2023 Enterovirus A71 (EV-A71) can elicit a wide variety of human diseases such as hand, foot, and mouth disease and severe or fatal neurological complications. It is not clearly understood what determines the virulence and fitness of EV-A71. It has been observed that amino acid changes in the receptor binding protein, VP1, resulting in viral binding to heparan sulfate proteoglycans (HSPGs) may be important for the ability of EV-A71 to infect neuronal tissue. In this study, we identified that the presence of glutamine, as opposed to glutamic acid, at VP1-145 is key for viral infection in a 2D human fetal intestinal model, consistent with previous findings in an airway organoid model. Moreover, pre-treatment of EV-A71 particles with low molecular weight heparin to block HSPG-binding significantly reduced the infectivity of two clinical EV-A71 isolates and viral mutants carrying glutamine at VP1-145. Our data indicates that mutations in VP1 leading to HSPG-binding enhances viral replication in the human gut. These mutations resulting in increased production of viral particles at the primary replication site could lead to a higher risk of subsequent neuroinfection.ImportanceWith the near eradication of polio worldwide, polio-like illness (as is increasingly caused by EV-A71 infections) is of emerging concern. EV-A71 is indeed the most neurotropic enterovirus that poses a major threat globally to public health and specifically in infants and young children. Our findings will contribute to the understanding of the virulence and the pathogenicity of this virus. Further, our data also supports the identification of potential therapeutic targets against severe EV-A71 infection especially among infants and young children. Furthermore, our work highlights the key role of HSPG-binding mutations in the disease outcome of EV-A71. Additionally, EV-A71 is not able to infect the gut (the primary replication site in humans) in traditionally used animal models. Thus, our research highlights the need for human-based models to study human viral infections.
Human milk inhibits some enveloped virus infections, including SARS-CoV-2, in an intestinal model Ikrame Aknouch, Adithya Sridhar, Eline Freeze, Francesca Paola Giugliano, Britt J van Keulen, et al. Life Science Alliance, 2022 Human milk is important for antimicrobial defense in infants and has well demonstrated antiviral activity. We evaluated the protective ability of human milk against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in a human fetal intestinal cell culture model. We found that, in this model, human milk blocks SARS-CoV-2 replication, irrespective of the presence of SARS-CoV-2 spike-specific antibodies. Complete inhibition of both enveloped Middle East respiratory syndrome coronavirus and human respiratory syncytial virus infections was also observed, whereas no inhibition of non-enveloped enterovirus A71 infection was seen. Transcriptome analysis after 24 h of the intestinal monolayers treated with human milk showed large transcriptomic changes from human milk treatment, and subsequent analysis suggested that ATP1A1 down-regulation by milk might be of importance. Inhibition of ATP1A1 blocked SARS-CoV-2 infection in our intestinal model, whereas no effect on EV-A71 infection was seen. Our data indicate that human milk has potent antiviral activity against particular (enveloped) viruses by potentially blocking the ATP1A1-mediated endocytic process.
Health-Related Quality of Life in Pediatric Inflammatory Bowel Disease During COVID-19 Pandemic: A Prospective Study Flora Fedele, Massimo Martinelli, Caterina Strisciuglio, Pasquale Dolce, Francesca Paola Giugliano, et al. Journal of Pediatric Gastroenterology and Nutrition, 2022 Objectives: The aims of our study were to evaluate health-related quality of life (HRQoL) in children affected by inflammatory bowel disease (IBD) during the first wave of Coronavirus disease 2019 (COVID-19) pandemic and after 12 months. Methods: This was a single-center, prospective, observational study conducted between April 2020 and April 2021. Children from 10 to 18 years with a confirmed diagnosis of IBD were enrolled during the first COVID-19–related national quarantine. The following information was collected at the baseline and after 12 months: IBD subtype, location and phenotype, disease activity, current and previous therapies. Patients were asked to complete the PROMIS Anxiety and IMPACT III questionnaires. Results: One hundred and eighteen patients were enrolled, of whom 54 (46%) were affected by Crohn disease (CD) and 64 (54%) with ulcerative colitis (UC; median age: 15.5 years, range 10.3–18; M/F: 68/50). Median HRQoL was significantly decreased after 12 months compared with the beginning of COVID-19–related quarantine (T1: 76.7 vs T2: 72.8; P < 0.001). At 12 months, a higher number of children were reported to be in active disease when compared with the enrollment [T2: 22/108 (20.4%) vs T1: 12/118 (10%); P = 0.02]. Multivariate analysis showed a significant influence on HRQoL of quarantine period (P < 0.001), female sex (P = 0.016), biologic therapy (P = 0.011), and active disease (P < 0.001). Conclusions: A deterioration of HRQoL after 12 months from COVID-19–related quarantine was observed. Additionally, the higher number of children with active disease at 12 months compared with enrollment may suggest detrimental consequences of the reduced disease control, contributing to decreased HRQoL.
Thiopurines correct the effects of autophagy impairment on intestinal healing - a potential role for ARHGAP18/RhoA Marileen M. C. Prins, Francesca P. Giugliano, Manon van Roest, Stan F. J. van de Graaf, Pim J. Koelink, et al. Dmm Disease Models and Mechanisms, 2021 The ATG16L1 T300A single-nucleotide polymorphism (SNP) is associated with Crohn's disease and causes an autophagy impairment. We have previously shown that this SNP is involved in the migration and hyperactivation of Rac1 in dendritic cells. Mucosal healing, currently the main target for inflammatory bowel disease treatment, depends on restoration of the epithelial barrier and requires appropriate migration of epithelial cells towards and over mucosal lesions. Therefore, we here further investigated the impact of autophagy on epithelial migration. ATG16L1 knockdown was established in the HT29 human colonic epithelial cell line using lentiviral transduction. Migratory capacity was evaluated using scratch assays and RhoAGTP was measured using G-LISA. Immunofluorescent ARHGAP18 and sequestome 1 (SQSTM1; also known as p62) staining was performed on HT29 cells and primary colonic tissue of Crohn's disease patients. We observed that ATG16L1 knockdown cells exhibited decreased autophagy and decreased migration capacity. Furthermore, activity of RhoA was decreased. These characteristics were phenocopied using ATG5 knockdown and pharmacological inhibition of autophagy. The migration defect was dependent on accumulation of SQSTM1 and was alleviated upon SQSTM1 knockdown. Strikingly, thiopurines also mitigated the effects of impaired autophagy. RhoA dysregulation appeared mediated through accumulation of the upstream regulator ARHGAP18, which was observed in cell lines, human foetal organoids and primary colonic tissue. Our results indicate that the ATG16L1 T300A Crohn's disease-associated SNP causes a decrease in migration capacity in epithelial cells, mediated by an increase in SQSTM1 and ARHGAP18 protein and subsequent reduced RhoA activation.
Disorders of Sucking and Swallowing Francesca Paola Giugliano, Erasmo Miele, Annamaria Staiano Textbook of Pediatric Gastroenterology Hepatology and Nutrition A Comprehensive Guide to Practice Second Edition, 2021
Disorders of Sucking and Swallowing Francesca Paola Giugliano, Erasmo Miele, Annamaria Staiano Textbook of Pediatric Gastroenterology Hepatology and Nutrition A Comprehensive Guide to Practice, 2015
