NMS: iNOVA4Health - Artigos em revista internacional com arbitragem científica
URI permanente para esta coleção:
Navegar
Entradas recentes
- mRNA delivery to the retina restores REP1 function in choroideremiaPublication . de Lemos, Luisa; Antas, Pedro; Carvalho, Cláudia; Castro, Mariana; Bruno, Diogo; Fonseca, Ana F; Rakshit, Shuvajit; Seabra, Miguel C; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); iNOVA4Health - pólo NMSInherited retinal diseases (IRDs) represent a major cause of blindness, yet current viral vector-based gene therapies are limited by high cost, restricted cargo capacity, and safety concerns related to immunogenicity. We explore here retinal delivery of non-viral, in-vitro-transcribed mRNA using lipid nanoparticles (LNPs) as a safe, cost-effective gene augmentation strategy for choroideremia (CHM). CHM is an X-linked IRD caused by loss-of-function mutations in CHM that encodes Rab escort protein-1 (REP1), which is essential for Rab GTPases prenylation and intracellular trafficking. We demonstrate that subretinal delivery of human CHM-mRNA (hCHM-mRNA) efficiently targets the retinal pigment epithelium (RPE) and choroid, achieving expression that is detectable up to 15 days post-injection, with lower and transient inflammatory response compared to AAV2 vectors. Furthermore, hCHM-mRNA delivery in vitro to human induced pluripotent stem cell (hiPSC)-derived RPE cells from a CHM patient and in vivo via subretinal injection in the Chm ± mouse model successfully restored REP1 levels and reversed Rab prenylation defect. Concordantly, Chm ± treated with hCHM-mRNA showed functional rescue of retinal activity measured by electroretinography 24 h post-injection. Overall, these results provide a strong proof-of-concept for LNP-mediated mRNA technology in the retina, supporting the development of scalable, non-viral gene approaches for IRDs, including CHM.
- Increased Expression of HLA-DR and CD69 on Peripheral CD4+ T Cells Predicts Better Clinical Outcomes in Cutaneous MelanomaPublication . Tomás, Ana; Maximino, José; Nunes, Hugo; Salvador, Rute; Luís, Rafael; Brito, Cheila; Saraiva, Diana P.; Gouveia, Emanuel; Pereira, Carolina; Gonçalves, Filipe; Farricha, Victor; Lopez Carvalho, Elisabete; Moura, Cecília; Passos, Maria José; Cristóvão-Ferreira, Sofia; Pereira, Patrícia M.; Cabral, Maria Guadalupe; Pojo, Marta; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); iNOVA4Health - pólo NMS; Dove Medical PressBackground: Cutaneous melanoma (CM) is an aggressive skin cancer with rising incidence, representing a growing public health concern. Despite the remarkable success of immune-checkpoint inhibitors (ICIs) in the management of advanced disease, mortality remains high due to therapy resistance. Identifying reliable prognostic and predictive biomarkers is therefore essential to improve patient stratification, optimize treatment selection, and minimize unnecessary toxicity. Methods: We comprehensively profiled the circulating immune landscape of 54 treatment-naïve CM patients by integrating flow cytometry immunophenotyping with clinicopathological data, and performed tumor gene expression analysis in a subset of 26 patients. Results: Elevated HLA-DR and CD69 expression on circulating CD4+ T cells, together with reduced circulating CD8+ T cell frequency, emerged as candidate prognostic biomarkers associated with improved survival. Prognostic models combining these immune variables with clinical covariates accurately stratified patients by overall survival (89.5% sensitivity, 72.7% specificity; AUC = 0.872, p < 0.0001) and progression/recurrence risk (75% sensitivity and 71.4% specificity; AUC = 0.763, p = 0.001). In a subset of 43 patients subsequently treated with ICIs, elevated baseline HLA-DR and CD69 expression on circulating CD4+ T cells was also associated with therapeutic benefit. A predictive model integrating these markers with clinical covariates achieved good discriminatory performance (65.2% sensitivity, 88.9% specificity; AUC = 0.775, p = 0.0027). Tumor gene expression profiling supported the role of IFN-γ-related signatures, previously linked to ICI response, as complementary prognostic and predictive tools. Conclusion: These findings highlight systemic CD4+ T cell activation status as a potential, easily measurable biomarker in CM, laying the foundation for future strategies to refine patient stratification and guiding immunotherapy decisions.
- Translational insights into extracellular vesicles in peritoneal dialysisPublication . Sequeira, Diogo; Anão, Sofia; Calça, Ana Rita; Matias, Patrícia João; de Melo Junior, Antonio Ferreira; Ferreira, João Vasco; Carvalho, Ana Sofia; Matthiesen, Rune; Branco, Patrícia; de Azeredo Pereira, Sofia; Sousa, Cátia; Teixeira-Santos, Luísa; iNOVA4Health - pólo NMS; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); Multimed Inc.Peritoneal dialysis (PD) is a cornerstone kidney replacement therapy for patients with end-stage kidney disease; however, chronic exposure to bioincompatible dialysis solutions progressively damages the peritoneal membrane, leading to mesothelial-to-mesenchymal transition, fibrosis, and ultimately ultrafiltration failure. Currently, peritoneal membrane dysfunction is detected only at advanced stages through the peritoneal equilibration test, underscoring the need for earlier and more sensitive biomarkers. Extracellular vesicles (EVs) isolated from peritoneal dialysis effluent (PDE) have emerged as promising candidates, given their capacity to carry proteins, lipids, and nucleic acids that reflect local and systemic cellular activity. To date, 12 studies have successfully isolated and characterized PDE-derived EVs, identifying canonical EV markers, such as CD9, CD63, CD81, TSG101, and HSP70, alongside disease-relevant molecules, including galectin-3 binding protein, aquaporin-1, glycoprotein 96, and integrin-linked kinase. These EVs are enriched in signaling components associated with inflammation, angiogenesis, and fibrosis, particularly through TGF-β/p38 and NF-κB pathways. Moreover, specific EV-associated microRNAs (e.g., miR-125a-5p, miR-132-3p, miR-296-3p, miR-432-5p) overlap with molecular signatures observed in kidney and cardiometabolic disorders, suggesting broader systemic relevance. As an original contribution to this review, and to address cross-study comparability, we applied EVqualityMS, a mass spectrometry-based quality assessment tool, to benchmark EV enrichment and contaminant profiles and calculate quality indices across publicly available PDE-EV proteomics datasets. Despite methodological heterogeneity and limited sample sizes, PDE-derived EVs represent a powerful “liquid biopsy” of the peritoneal environment. Their integration into PD monitoring holds promise for the early detection of membrane injury and for supporting a predictive, biomarker-guided, and personalized approach to PD management.
- Development of a predictive model for postoperative body mass index and diabetes outcomes after metabolic bariatric surgeryPublication . Ochs, Vincent; Kollmann, Lars; Rosenblum, Ilan; Poljo, Adisa; Heule, Andreas; Enodien, Bassey; Chumakova-Orin, Maryna; DeMaria, Eric J.; Burri, Emanuel; Stoll, Reinhard; Kollmar, Otto; Rosenberg, Robert; Probst, Pascal; Muller, Markus K.; Taha-Mehlitz, Stephanie; Müller, Beat P.; Frey, Daniel M.; Kalinowski, Piotr; Przybysz, Marta; Bartkowiak, Mateusz; Dalkilic, Muhammed Said; Sisik, Abdullah; Oliveira, Rodrigo Otavio Carvalho de; Martins, Fatima; Stenberg, Erik; Andersson, Ellen; Olbers, Torsten; Flemming, Sven; Seyfried, Florian; Ponholzer, Florian; Weissenbacher, Annemarie; Öfner, Dietmar; Betzler, Johanna; Otto, Mirko; Peterli, Ralph; Cattin, Philippe C.; Taha, Anas; iNOVA4Health - pólo NMS; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); Oxford University PressBackground: Predicting postoperative body mass index (BMI) trajectories and long-term type 2 diabetes (T2D) remission after bariatric surgery remains challenging. Existing models often rely on baseline variables only and fail to incorporate dynamic postoperative changes. This study aimed to develop and validate a multicentre machine-learning framework that predicts individualized BMI trajectories and T2D remission using routinely available preoperative data and time-dependent weight evolution. Methods: This multicentre retrospective cohort study included adult patients who underwent Roux-en-Y gastric bypass or sleeve gastrectomy across 11 European centres (2012–2023). Variables with > 30% missing data were excluded; remaining missing values were imputed iteratively. A two-stage approach was used: a regression model predicting postoperative BMI at 3–60 months using an autoregressive design; and a classification model predicting T2D remission using baseline features and predicted BMI trajectories. Internal performance was evaluated with ten-fold and leave-one-clinic-out cross-validation; external validation used an independent cohort from Linköping, Sweden. Results: Of the 11 457 patients initially identified, 9652 patients with complete baseline and follow-up information were used for the analysis. The best BMI model (HistGradientBoosting) achieved a root mean square error (RMSE) of 1.11 kg/m2 (95% confidence interval 1.07 to 1.14) and a mean absolute error (MAE) of 0.62 kg/m2 across clinics; external testing showed an RMSE of 1.12 kg/m2 (95% confidence interval 1.11 to 1.12) and an MAE of 0.63 kg/m2. The T2D remission classifier (XGBoost) obtained a Macro F1 score of 0.88 (precision 0.87, recall 0.88), with an external F1 score of 0.89. Incorporating predicted BMI trajectories improved discrimination compared with baseline-only models (C-index 0.95 versus 0.93). Conclusion: A two-stage machine-learning framework has high predictive performance for postoperative BMI and T2D remission up to 5 years after bariatric surgery. Dynamic incorporation of predicted weight trajectories enhances metabolic risk prediction and supports individualized counselling and postoperative management.
- Pre-Transplant Serum FTIRS Signatures as Predictive Biomarkers of Early Transient Pancreatic Graft Dysfunction in Simultaneous Pancreas-Kidney TransplantationPublication . Vigia, Emanuel; Ramalhete, Luís; Araújo, Rúben; Corado, Sofia; Barros, Inês; Chumbinho, Beatriz; Nobre, Ana; Carrelha, Sofia; Pico, Paula; Rodrigues, Fernando; Vieira, Miguel Bigotte; Magriço, Rita; Cotovio, Patrícia; Caeiro, Fernando; Aires, Inês; Silva, Cecília; Pena, Ana; Bicho, Luís; Jorge, Cristina; Calado, Cecília R.C.; Pereira, Jorge P.; Ferreira, Aníbal; Marques, Hugo P.; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); iNOVA4Health - pólo NMS; MDPI - Multidisciplinary Digital Publishing InstituteBackground/Objectives: Early transient endocrine dysfunction after simultaneous pancreas-kidney transplantation (SPK) frequently triggers urgent investigations to exclude thrombosis, pancreatitis, or rejection, yet many recipients recover during the index admission. We tested whether pre-transplant day zero (D0) serum Fourier-transform infrared spectroscopy (FTIRS) captures a biochemical fingerprint associated with a Start&Stop trajectory (initial insulin independence followed by transient dysfunction with recovery). Methods: In a single-center retrospective case-control study nested within 104 consecutive SPK recipients with available D0 serum, 12 Start&Stop cases were matched 1:1 to 12 No-Stop controls. Serum FTIR spectra went through structured quality control and standardized preprocessing. A Naïve Bayes classifier with Fast Correlation-Based Filter (FCBF) feature selection was evaluated using leave-one-out cross-validation (LOOCV) and label-permutation analysis. Results: Under LOOCV, the primary FTIRS model (Savitzky-Golay second derivative; 600–900 and 2800–3400 cm−1) achieved excellent discrimination (ROC-AUC 1.00) with accuracy 0.958 and F1 score 0.958. Discrimination collapsed under label permutation (ROC-AUC 0.461), supporting a non-random label-spectrum association. Discriminant information mapped mainly to carbohydrate/glycoprotein-associated bands (~946–1161 cm−1), protein structural contributions near the amide III region (~1300 cm−1), and lipid/protein stretching modes (~2865–3163 cm−1), consistent with a multicomponent systemic biochemical state. Conclusions: In this exploratory matched case-control cohort, pre-transplant D0 serum FTIRS signatures were associated with the subsequent Start&Stop phenotype after SPK. These findings should be interpreted as recipient-side exploratory risk-stratification signals rather than clinically actionable decision tools. Larger multicenter validation in unselected cohorts, with standardized endpoint adjudication, preanalytical control, fully nested model development and inter-instrument harmonization, is required before clinical implementation or population-level risk calibration.
- Metabolic drivers of MASLD and MASHPublication . Gastaldelli, Amalia; Scoditti, Egeria; Macedo, Maria Paula; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); iNOVA4Health - pólo NMS; SpringerMetabolic dysfunction-associated steatotic liver disease (MASLD) and its more severe and aggressive form, metabolic dysfunction-associated steatohepatitis (MASH), represent liver-specific consequences of systemic metabolic dysfunction. This review synthesises current evidence on the metabolic and hormonal determinants of MASLD progression, with a particular focus on the mechanisms linking systemic dysfunction across multiple tissues to hepatic inflammation and fibrosis. MASLD pathogenesis is driven by complex interactions between insulin resistance, impaired glucose metabolism, dysfunctional adipose tissue lipolysis and altered hepatic de novo lipogenesis. Central to this process is the dysregulation of key metabolic hormones, including insulin and glucagon, whose imbalance disrupts both glucose and lipid fluxes, leading not only to hepatic steatosis but also to altered hepatic glucose handling, increased substrate delivery and reduced metabolic flexibility. This promotes lipid accumulation as well as glucotoxic and lipotoxic stress. These metabolic disturbances trigger hepatocellular injury and activate inflammatory signalling pathways, including adipokine-mediated crosstalk between adipose tissue and the liver, promoting hepatic stellate cell activation and ultimately leading to a state of chronic low-grade inflammation (metaflammation) that integrates hepatocyte stress, immune cell activation and hepatic stellate cell-driven fibrogenesis. This review also highlights sex-specific hormonal regulation and the gut–liver–adipose axis as determinants of disease heterogeneity and immunometabolic injury. By integrating mechanistic insights across glucose and lipid metabolism, tissues and hormonal pathways, this review underscores how systemic metabolic dysfunction is translated into progressive liver injury and fibrosis. Understanding these interconnected mechanisms of disease progression is essential for identifying therapeutic targets and advancing mechanism-based and precision medicine approaches for MASLD and MASH.
- Socioeconomic burden of osteoarthritis in Sub-Saharan Africa and the determinants of healthcare inequitiesPublication . Andela, Leonel; Andrade, Carlos Augusto Ferreira de; Souza, Breno Augusto Bormann de; Filho, Augusto Bormann de Souza; Pimentel-Santos, Fernando; Ferreira, Daniele Masteson Tavares Pereira; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); iNOVA4Health - pólo NMS; BMJ Publishing GroupINTRODUCTION: Access to musculoskeletal healthcare services in Sub-Saharan Africa is inadequate. As osteoarthritis is the most prevalent chronic osteoarticular disease globally, it's essential to understand its social and economic impact, as well as the determinants of inequities in access to healthcare services in Sub-Saharan Africa. The absence of systematised knowledge on this topic makes this review pertinent. However, due to data scarcity, assessing this burden is challenging. The objective of this scoping review is to map and summarise the available literature up to 2025 on the socioeconomic burden and health inequity determinants among the Sub-Saharan African population with osteoarthritis. METHODS AND ANALYSIS: A predefined search strategy will be applied to MEDLINE (via PubMed), Embase, African Journals Online and African Index Medicus to incorporate articles relevant to adults diagnosed with osteoarthritis who are residents of sub-Saharan Africa. We will also include grey literature sources such as Google Scholar, Research Square, manuals, books, medical society websites, secondary databases, theses and dissertation repositories and conference proceedings. Study selection will be conducted in two stages by a pair of reviewers who will independently screen titles and abstracts according to the eligibility criteria, followed by a full-text review of the selected studies. The search period was from October 2025 to January 2026. Data extraction will be performed using a standardised charting form developed by the review team. ETHICS AND DISSEMINATION: This scoping review maps evidence on OA-related socioeconomic impacts and healthcare inequities in Sub-Saharan Africa. As a secondary data analysis, ethical approval is not required. Findings will be disseminated via peer-reviewed journals and academic conferences to clinicians and policymakers.
- Protocol to isolate endosomal and small extracellular vesicles from cultured cells through ultracentrifugationPublication . Domingues, Maria Nolasco; Palhinhas, Luís; Pereira, Paulo; Ferreira, João Vasco; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); iNOVA4Health - pólo NMS; Cell PressHere, we present a protocol to isolate endosomal fractions using sucrose-density gradient ultracentrifugation and to recover small extracellular vesicles (sEVs), enriched in exosomes, using sequential ultracentrifugation from mammalian cell lines. This combined approach enables the separation and analysis of early endosome (EE), late endosome (LE), and sEV fractions. We provide detailed procedures for cell culture preparation, conditioned media collection, differential centrifugation, gradient layering, and fraction purification, facilitating downstream characterization and functional assays. For complete details on the use and execution of this protocol, please refer to Ferreira et al.
- mRNA therapy improves the composition and motility in CCDC40-deficient cilia in vitro and in vivoPublication . Wohlgemuth, Kai; Rasteiro, Margarida; Aneja, Manish; Bota, Catarina; Cindric, Sandra; Freischem, Stefanie; George, Sebastian; Günsel, Gizem Günes; Ishola, Seun; Koenig, Julia; Kubisch-Dohmen, Rebekka; Langenickel, Thomas; Loges, Niki Tomas; Lopes, Miguel; Mummert, Verena; Olbrich, Heike; Pennekamp, Petra; Pereira, Telmo; Pinto, Andreia L; Raidt, Johanna; Rudolph, Carsten; Ter Steege, Adrian; Valecha, Drishti; Lopes, Susana S; Omran, Heymut; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); iNOVA4Health - pólo NMS; Faculdade de Ciências e Tecnologia (FCT); DCV - Departamento de Ciências da Vida; American Thoracic SocietyPrimary Ciliary Dyskinesia (PCD) is a genetically heterogeneous disorder leading to destructive airway disease with severe bronchiectasis and chronic lung failure in adulthood. Pathogenic variants in CCDC40 are associated with more severe reduction of lung function compared to most other PCD types. Currently, no therapies correcting the underlying disease mechanism are available. Here we investigate the efficacy of lipidoid nanoparticle-formulated mRNA encoding human CCDC40 (LNP-CCDC40-mRNA) as a corrective measure for structural and functional defects in vitro (human cells) and in vivo (zebrafish). Human nasal respiratory epithelial cells cultured at air-liquid-interface from five CCDC40-deficient individuals and a newly generated vertebrate animal model (ccdc40-/- zebrafish) were treated with LNP-CCDC40-mRNA. CCDC40-deficient cells were analyzed by high-speed video microscopy and immunofluorescence microscopy. ccdc40-/- zebrafish olfactory pit cilia were analyzed by high-speed video microscopy and fluid flow assays. Topical application of exogenous LNP-CCDC40-mRNA to CCDC40-deficient cells results in endogenous CCDC40 expression (10-74% of ciliated cells), enabling axonemal integration of CCDC40-associated proteins (CCDC39, GAS8/DRC4, DNALI1). Consistently, ciliary beat frequencies were significantly increased in treated CCDC40-deficient cells and comparable to healthy control cells. Further, we showed improved ciliary transport of fluorescent particles. Injection or topical application of human LNP-CCDC40-mRNA to ccdc40-/- zebrafish significantly increased ciliary motility and established directional flow in olfactory pits. We provide structural and functional evidence in vitro and in vivo for the biological efficacy of LNP-CCDC40-mRNA in CCDC40-deficient respiratory cells and zebrafish. Based on our results, an in vivo human study (Phase 1 trial) is planned in individuals with pathogenic variants in CCDC40.
- Urinary bowel stonePublication . Santos, Diana; Navarro, David; Jorge, Cristina; Fonseca, Nuno Moreira; iNOVA4Health - pólo NMS; NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM); BioMed Central (BMC)BACKGROUND: Urinary diversion with an ileal conduit is associated with an increased risk of urinary tract calculi, driven by metabolic disturbances, recurrent infections, urinary stasis, and foreign bodies. Although stone formation is a known complication, calculi developing around migrated surgical material remain infrequently reported. CASE PRESENTATION: An 81-year-old male patient with a history of radical cystectomy and ileal conduit formation for invasive urothelial carcinoma presented with flank pain and urinary tract infection four years after surgery. Morphological analysis of two spontaneously expelled urinary stones retrieved from the conduit drainage bag classified both calculi as type IVc, with a central nucleus composed of surgical staples. Fourier-transform infrared spectroscopy revealed a mixed composition of struvite (80%) and sodium hydrogenurate (20%), consistent with infection-related lithiasis. The findings supported stone formation around migrated foreign material in the context of chronic bacteriuria. CONCLUSION: Surgical staple migration can act as a nidus for infection-related stone formation in patients with ileal conduits, even several years after cystectomy. Stone analysis plays a crucial role in identifying foreign-body-associated calculi and possible underlying mechanisms. Clinicians should maintain a high index of suspicion for urolithiasis in patients with urinary diversions presenting with flank pain or recurrent infections. CLINICAL TRIAL REGISTRATION: Not applicable.