IRIS UniSR (’Università Vita-Salute San Raffaele)
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Spatiotemporal liver dynamics shape hepatocellular heterogeneity and impact in vivo gene engineering
No full textBackground & Aims: Hepatocytes are the liver's main functional cells and are key targets for in vivo gene therapy to treat monogenic diseases. Integrating the transgene into the genome is critical for long-term expression from a single early-life dose, which is achievable via integrating vectors or genome editing. To ensure persistence through liver growth and cell turnover, it is also necessary to target the hepatocytes driving these processes. While liver regeneration and homeostasis have been studied extensively, hepatocyte growth and maturation remain less well understood. Here, we investigate how hepatocyte heterogeneity evolves during liver growth and its implications for in vivo gene engineering. Methods: We performed clonal tracing, as well as single-cell and spatial transcriptomics, on mouse livers of various ages. We evaluated the efficiency, stability, and lobule distribution of lentiviral gene transfer and targeted transgene integration. Results: We found that a subset of clonogenic hepatocytes (15-20%) in the newborn liver generates >90% of the adult tissue and co-localizes with hematopoietic islands within a spatial niche. Preferential gene editing of these clonogenic hepatocytes resulted in an increased proportion of the gene-engineered liver area, supporting their role in liver growth. Age-dependent hepatocellular heterogeneity affected the efficiency of lentiviral gene delivery in vivo and its distribution throughout the hepatic lobule. The gradual establishment of metabolic zonation after weaning and elevated proteasome activity in the peri-central area in adults influenced the observed age-related outcomes. Conclusion: These insights into spatiotemporal hepatocyte dynamics enhance our understanding of liver biology and have important implications for therapeutic strategies. Impact and implications: We provide new insights into the spatiotemporal dynamics of the mouse liver during postnatal growth, highlighting both proliferative and transcriptomic heterogeneity among hepatocytes and their impact on the efficiency and distribution of in vivo lentiviral gene delivery and targeted gene editing. Understanding and manipulating the biological processes behind this heterogeneity can enhance gene transfer outcomes. We report that not all hepatocytes contribute equally to liver growth, indicating that effectively targeting clonogenic hepatocytes in the newborn liver is crucial for the long-term maintenance of therapeutic genetic modifications. Furthermore, this phenomenon can be leveraged to expand the pool of genetically corrected cells, as illustrated here by a targeted gene editing strategy. Finally, we reveal the existence of a tissue niche that supports the proliferation of both clonogenic hepatocytes and hematopoietic progenitors in neonatal livers. Gaining a deeper understanding of this niche and its signals may be beneficial for regenerative purposes
Prognostic Validation of Perineural Invasion Severity Score in Pancreatic Cancer: A Prospective Study
No full textObjective: This study aims to prospectively validate a perineural invasion (PNI) severity score in predicting recurrence and survival in resected PDAC. Background: PNI in pancreatic ductal adenocarcinoma (PDAC) is characterized by clinical and morphologic heterogeneity. Although a specific score for PNI severity has been proposed, it lacks validation. Methods: In this prospective registered (NCT04024358) monocentric study, 300 patients undergoing pancreatectomy between March 2019 and February 2022 were analyzed. PNI was scored as: 0 absent; 1 presence of neoplasia along nerves<3 mm; 2 neoplastic infiltration of nerves≥3 mm and/or massive PNI and/or necrosis of the infiltrated nerves. Association of PNI with disease recurrence and survival was evaluated. Results: Of 300 patients, 86% presented PNI, with 148 (49.3%) having PNI 1 and 110 (36.7%) PNI 2. Neoadjuvant treatment did not influence PNI (89.7% vs. 83.9% after treatment, P=0.332). PNI severity significantly correlated with worsening pathological features, shorter disease recurrence (24 mo for PNI 0, 17 for PNI 1 and 15 for PNI 2, P<0.01) and survival (57 mo for PNI 0, 51 for PNI 1 and 32 for PNI 2, P<0.01). PNI 2 independently predicted both recurrence and survival with HR of 2.082 (P=0.006) and 3.304 (P=0.014). PNI 2 benefitted most from adjuvant treatment, with longer disease recurrence time (17 vs. 12 mo; P=0.007) and survival (36 vs. 17 mo; P=0.004). Conclusions: The PNI severity score improves prognostic stratification of resected PDAC and identifies patients who benefit the most from adjuvant treatment
EARLY RECOGNITION AND PROMPT TREATMENT OF ACUTE SARCOPENIA: IMPACT ON SHORT- AND LONG-TERM OUTCOMES
No full textDevelopment and Validation of a Predictive Risk Score for Blood Transfusion in Patients Undergoing Curative-Intent Surgery for Intrahepatic Cholangiocarcinoma
Get PDFBackground and Objectives: Among patients undergoing liver resection for intrahepatic cholangiocarcinoma (ICC), perioperative bleeding requiring blood transfusion is a common complication, yet preoperative identification of patients at risk for transfusion remains challenging. The objective of this study was to develop a preoperative risk score for blood transfusion requirement during surgery for ICC. Methods: Patients undergoing curative-intent liver surgery for ICC (1990–2020) were identified from a multi-institutional database. A predictive model was developed and validated. An easy-to-use risk calculator was made available online. Results: Among 1420 patients, 300 (21.1%) received an intraoperative transfusion. Independent predictors of transfusion included severe preoperative anemia (OR = 1.65, 95% CI 1.10–2.47), T2 category or higher (OR = 2.00, 95% CI 1.36–3.02), positive lymph nodes (OR = 1.75, 95% CI 1.32–2.32) and major resection (OR = 2.56, 95%CI 1.85–3.58). Receipt of blood transfusion significantly correlated with worse outcomes. The model showed good discriminative ability in both training (AUC = 0.68, 95% CI 0.66–0.72) and bootstrapping validation (C-index = 0.67, 95% CI 0.65–0.70) cohorts. An online risk calculator of blood transfusion requirement was developed (https://catalano-giovanni.shinyapps.io/TransfusionRisk). Conclusions: Intraoperative blood transfusion was significantly associated with poor postoperative outcomes among patients undergoing surgery for ICC. The identification of patients at high risk of transfusion could improve perioperative patient care and blood resources allocation
Comparative Analysis of Islet Auto-Transplantation Outcome Classification Systems: Evaluating Concordance, Feasibility, and a Data-Driven Approach
No full textA standardized approach to assessing islet autotransplantation outcomes is crucial for evaluating graft function and guiding clinical decisions. This study compares the performance of existing classification systems—Milan, Minneapolis, Chicago, Leicester, Igls, and a novel Data-Driven approach—by evaluating their ability to differentiate transplant outcomes using metabolic and insulin secretion parameters. Our analysis shows strong concordance among Milan, Minneapolis, Chicago, and Igls, primarily due to minor variations in C-peptide thresholds. The Leicester and Data-Driven systems, however, exhibit greater divergence, with the Leicester system simplifying assessment by excluding severe hypoglycemic events and HbA1c, and the Data-Driven approach offering a more dynamic framework without predefined thresholds. Fasting C-peptide levels emerged as a highly reliable predictor of graft function, with the arginine test proving more effective than Mixed Meal Tolerance Test for additional evaluation. The Data-Driven approach provided superior stratification of outcomes, highlighting the importance of residual insulin secretion in metabolic control. These findings suggest that refining classification systems, particularly by considering insulin sensitivity and residual secretion, could enhance long-term patient monitoring and improve our understanding of beta-cell replacement therapies. Further validation across diverse cohorts is essential for broader clinical adoption
Hemodynamic Right Heart Catheterization before Transcatheter Mitral and Tricuspid Therapies
No full textRecent findings emphasize the potential role of invasive hemodynamic assessment in guiding transcatheter mitral and tricuspid valve percutaneous interventions. Right heart catheterization-derived parameters offer insights into hemodynamic changes associated with valvular heart diseases, pulmonary hypertension phenotyping, and right ventricular to pulmonary artery coupling. This might improve prognostic stratification for candidates to transcatheter therapies. This review provides a clinical overview of available data regarding the utility of preoperative right heart catheterization-derived parameters in patients undergoing mitral and tricuspid percutaneous repair or replacement
Postvaccine early-onset (PoVEO) forms of Graves' disease after anti-SARS-CoV-2 vaccination are characterized by favourable long-term disease outcomes
No full textIntroduction: In a previous cohort of patients with new-onset of Graves' Disease (GD) during 2021, a "postvaccine early-onset (PoVEO)" after anti-COVID-19 vaccination emerged as a new and frequent nosological entity occurring in approximately one third of patients. Similar cohorts have also been reported with GD after anti-COVID vaccines, but to date there are no available data on long-term clinical outcomes. Methods: This is a single-center retrospective study evaluating the 24-month clinical course of patients with early-onset (<4 weeks) GD after anti-COVID vaccination (PoVEO) compared to non-PoVEO patients. Biochemical tests, therapies used and follow-up visits were scheduled according to current guidelines, clinicians' experience and patients' needs. Results: As previously described, 64 patients were observed in 2021 and 20 (31.2%) had PoVEO. Individuals with PoVEO were characterized by distinctive features such as older age, a higher prevalence of male sex, and a better initial biochemical response. Eleven individuals were followed at other Centers (2 with PoVEO and 9 without) and were lost to follow-up. During the two-year follow-up, of the 53 individuals, 7 (13%) underwent definitive surgical or radioactive-iodine therapy (3 PoVEO and 4 non-PoVEO p = 0.67), and 30 (57%) presented disease remission with medical therapy alone. Excluding those who underwent definitive treatments, remission with medical therapy alone was observed in 13/15 (87%) individuals with PoVEO form and 17/31 (55%) non-PoVEO (p = 0.048). Conclusion: In this retrospective study of a cohort of individuals with the PoVEO form of GD, we found a more favourable clinical course, likely linked to transience of the triggering events. Despite these findings, individuals presenting with GD after SARS-CoV-2 vaccination should undergo an active surveillance program with repeat thyroid function testing
Quality of life in heart failure. The heart of the matter. A scientific statement of the Heart Failure Association and the European Association of Preventive Cardiology of the European Society of Cardiology
No full textFor most patients with chronic, progressive illnesses, maintaining good quality of life (QoL), with preserved functional capacity, is just as crucial as prolonging survival. Patients with heart failure (HF) experience much worse QoL and effort intolerance than both the general population and people with other chronic conditions, since they present a range of physical and psychological symptoms, including shortness of breath, chest discomfort, fatigue, fluid congestion, trouble with sleeping, and depression. These symptoms reduce patients' capacity for daily social and physical activity. Usual endpoints of large-scale trials in chronic HF have mostly been defined to evaluate treatments regarding hospitalizations and mortality, but more recently, patients' priorities and needs expressed with QoL are gaining more awareness and are being more extensively evaluated. This scientific statement aims at discussing the importance of QoL in HF, summarizing the most largely adopted questionnaires in HF care, and providing an overview on their application in trials and the potential for their transition to clinical practice. Finally, by discussing the reasons limiting their application in daily clinical routine and the strategies that may promote their implementation, this statement aims at fostering the systematic integration of the patient's standpoint in HF care
Neonatal gene therapy effectively prevents disease manifestations in a murine model of Mucopolysaccharidosis type I
No full textMucopolysaccharidosis type I (MPS-I) is a rare pediatric disease caused by mutations in the α-L-iduronidase (IDUA) gene encoding for a lysosomal enzyme involved in glycosaminoglycan metabolism. While newborns with the severe Hurler variant are usually asymptomatic at birth, progressive disease manifestations emerge early in life. Since previous studies on lentiviral vector gene therapy (GT) in Hurler patients have demonstrated superior metabolic correction and early beneficial clinical effects, we investigated whether applying this GT approach during the neonatal period could be effective in preventing disease pathology before it becomes irreversible. Thus, newborn MPS-I mice were transplanted with affected bone marrow-derived progenitor cells transduced with an IDUA-encoding lentiviral vector. Treated animals displayed increased IDUA levels, significantly reducing substrate accumulation in analyzed organs, indicating metabolic correction. Skeletal manifestations, typically resistant to conventional therapies, showed improvements at radiographic and histological levels post-treatment. Additionally, a decrease in brain cortex vacuolization and inflammation suggested neurological amelioration. Overall, this study provides a proof of principle demonstrating the effectiveness of neonatal ex vivo GT in MPS-I mice and supports its potential for further optimization at the pre-clinical level
Robotic Liver Resection for Hepatocellular Carcinoma: A Multicenter Case Series
No full textBackground: Liver resection is the standard treatment for resectable hepatocellular carcinoma (HCC). The advent of robotic surgery has extended its application in liver surgery, reducing post-operative complications without compromising oncological safety. This study is a retrospective series with the aim of analyzing the preoperative patient’s and tumor’s characteristics and evaluating intraoperative and post-operative data in terms of hospital stay, complications, and oncological radicality. Methods: Data were collected from a multicenter retrospective database that includes 1070 consecutive robotic liver resections (RLRs) performed in nine European hospital centers from 2011 to 2023. Of the entire series, 343 liver resections were performed for HCC. Results: A total of 247 patients (72.3%) had mono-focal lesions. Major hepatectomies and anatomical resections have been perfomed in 87% and 55% of patients, respectively. All 17 conversions (4.95%) were to the open approach. The operative mean time was 239.56 min and the estimated blood loss was 229.45 mL. The overall post-operative complication rate was 22.74%, but severe complications occurred in 4.08% of patients and one of them (0.29%) was reoperated on. The mean hospital stay was 5.82 days with a mean ICU stay of 0.9 days. Twenty-six resections (7.6%) were R1 parenchymal. Forty-six patients (4.08%) were readmitted to the hospital within 90 days after discharge and seventy-eight patients (22.74%) had disease recurrence. Total deaths included 36 (10.5%) patients with a 90-day mortality of 0.9%. Conclusions: Robotic liver resection for HCC is feasible and safe when performed in experienced centers by surgeons who have completed the learning curve