Pharmacology (medical), Pathology and Forensic Medicine, Pediatrics
13
Scopus Publications
Scopus Publications
Development and application of a multimatrix LC–MS/MS method for quantifying elexacaftor–tezacaftor–ivacaftor: Expanding therapeutic drug monitoring in cystic fibrosis from systemic circulation to airways and sweat Matteo Mucci, Martina Colarelli, Pietro Ripani, Marta Di Nicola, Marianna Del Ciotto, Maria Di Sabatino, Francesca Collini, Mario Romano, Antonio Recchiuti Biomedicine and Pharmacotherapy, 2025 Therapeutic drug monitoring (TDM) is becoming increasingly essential in cystic fibrosis (CF), as a growing number of children and adults are now eligible for elexacaftor/tezacaftor/ivacaftor (ETI) CF Transmembrane conductance Regulator (CFTR) modulator therapies. Although plasma remains the benchmark, its invasive nature limits practical use. Analytical approaches based on dried matrices currently available still rely on venipuncture and intensive workflows. To overcome these limitations, we developed an advanced, multidimensional TDM framework for ETI quantification that incorporates truly venipuncture-free, self-collected, and quantitative dried blood spot (DBS) sampling, along with non-invasive nasal airway swabs (NAS) and sweat. We also evaluated ETI in airway and sweat fluids, offering insight into drug distribution and activity at key target tissues. Selectivity, specificity, linearity, accuracy, precision, and inter/intraday stability of the methods applied to plasma, NAS, and sweat specimens were investigated according to ICH M10 guidelines and IATDMCT for DBS. Analytical performance of the methods in all tested matrices were demonstrated and met the acceptance criteria of ICH M10 guidelines for bioanalytical-method validation, with DBS additionally fulfilling the IATDMCT DBS-specific recommendations. DBS measurements were statistically equivalent to plasma concentrations, supporting their utility as a minimally invasive surrogate for TDM. ETI levels in NAS were significantly higher than in plasma or sweat, indicating localized accumulation of the drug on the airway surface, a key site of CFTR activity. Sweat samples, while showing lower analyte amounts, contained detectable levels in all analytes within their biologically active concentrations. This integrated analytical approach provides a holistic view of systemic and local ETI distribution, increasing the potential for personalized TDM, pharmacokinetics/pharmacodynamics study, and optimization of CFTR modulator therapy. Created in BioRender. Recchiuti, A. (2025) https://BioRender.com/n2zhuv1 • Monitoring CFTR modulator drug concentrations in people with cystic fibrosis is increasingly required. • Dried blood spots enable accurate systemic monitoring without venipuncture. • Nasal swabs can be used to measure high airway concentrations of modulator drugs. • Sweat sampling offers a new window into drug exposure and CFTR activity. • The multimatrix approach advances holistic precision monitoring in cystic fibrosis.
Inflammation and epithelial–mesenchymal transition in a CFTR-depleted human bronchial epithelial cell line revealed by proteomics and human organ-on-a-chip Domenico Mattoscio, Luis A. Baeza, Haiqing Bai, Tommaso Colangelo, Simone Castagnozzi, Marta Marzotto, Maria Concetta Cufaro, Virginia Lotti, Yu‐Chieh Yuan, Matteo Mucci, Longlong Si, Mariachiara Zuccarini, Maria Tredicine, Simona D'Orazio, Damiana Pieragostino, Piero Del Boccio, Claudio Sorio, Marco Trerotola, Mario Romano, Roberto Plebani FEBS Journal, 2025 Cystic fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene, leading to chronic, unresolved inflammation of the airways due to uncontrolled recruitment of polymorphonuclear leukocytes (PMNs). Evidence indicates that CFTR loss‐of‐function, in addition to promoting a pro‐inflammatory phenotype, is associated with an increased risk of developing cancer, suggesting that CFTR can exert tumor‐suppressor functions. Three‐dimensional (3D) in vitro culture models, such as the CF lung airway‐on‐a‐chip, can be suitable for studying PMN recruitment, as well as events of cancerogenesis, that is epithelial cell invasion and migration, in CF. To gather insight into the pathobiology of CFTR loss‐of‐function, we generated CFTR‐knockout (KO) clones of the 16HBE14o‐ human bronchial cell line by CRISPR/Cas9 gene editing, and performed a comparative proteomic analysis of these clones with their wild‐type (WT) counterparts. Systematic signaling pathway analysis of CFTR‐KO clones revealed modulation of inflammation, PMN recruitment, epithelial cell migration, and epithelial–mesenchymal transition. Using a latest‐generation organ‐on‐a‐chip microfluidic platform, we confirmed that CFTR‐KO enhanced PMN recruitment and epithelial cell invasion of the endothelial layer. Thus, a dysfunctional CFTR affects multiple pathways in the airway epithelium that ultimately contribute to sustained inflammation and cancerogenesis in CF.
Immunoregulatory mechanisms of the arachidonic acid pathway in cancer Maria Tredicine, Matteo Mucci, Antonio Recchiuti, Domenico Mattoscio FEBS Letters, 2025 The arachidonic acid (AA) pathway promotes tumor progression by modulating the complex interactions between cancer and immune cells within the microenvironment. In this Review, we summarize the knowledge acquired thus far concerning the intricate mechanisms through which eicosanoids either promote or suppress the antitumor immune response. In addition, we will discuss the impact of eicosanoids on immune cells and how they affect responsiveness to immunotherapy, as well as potential strategies for manipulating the AA pathway to improve anticancer immunotherapy. Understanding the molecular pathways and mechanisms underlying the role played by AA and its metabolites in tumor progression may contribute to the development of more effective anticancer immunotherapies.
Resolvin D1 improves airway inflammation and exercise capacity in cystic fibrosis lung disease Giulia Ferri, Matteo Serano, Elisa Isopi, Matteo Mucci, Domenico Mattoscio, Romina Pecce, Feliciano Protasi, Marcus A. Mall, Mario Romano, Antonio Recchiuti FASEB Journal, 2023 Mucus plugging and non‐resolving inflammation are inherent features of cystic fibrosis (CF) that may lead to progressive lung disease and exercise intolerance, which are the main causes of morbidity and mortality for people with CF. Therefore, understanding the influence of mucus on basic mechanisms underlying the inflammatory response and identifying strategies to resolve mucus‐driven airway inflammation and consequent morbidity in CF are of wide interest. Here, we investigated the effects of the proresolving lipid mediator resolvin (Rv) D1 on mucus‐related inflammation as a proof‐of‐concept to alleviate the burden of lung disease and restore exercise intolerance in CF. We tested the effects of RvD1 on inflammatory responses of human organotypic airways and leukocytes to CF mucus and of humanized mice expressing the epithelial Na + channel (βENaC‐Tg) having CF‐like mucus obstruction, lung disease, and physical exercise intolerance. RvD1 reduced pathogenic phenotypes of CF‐airway supernatant (ASN)‐stimulated human neutrophils, including loss of L‐selectin shedding and CD16. RNASeq analysis identified select transcripts and pathways regulated by RvD1 in ASN‐stimulated CF bronchial epithelial cells that are involved in sugar metabolism, NF‐κB activation and inflammation, and response to stress. In in vivo inflammation using βENaC TG mice, RvD1 reduced total leukocytes, PMN, and interstitial Siglec‐MΦ when given at 6–8 weeks of age, and in older mice at 10–12 weeks of age, along with the decrease of pro‐inflammatory chemokines and increase of anti‐inflammatory IL‐10. Furthermore, RvD1 treatment promoted the resolution of pulmonary exacerbation caused by Pseudomonas aeruginosa infection and significantly enhanced physical activity and energy expenditure associated with mucus obstruction, which was impaired in βENaC‐Tg mice compared with wild‐type. These results demonstrate that RvD1 can rectify features of CF and offer proof‐of‐concept for its therapeutic application in this and other muco‐obstructive lung diseases.
Specialized pro-resolving lipid mediators and resolution of viral diseases Giulia Ferri, Matteo Mucci, Domenico Mattoscio, Antonio Recchiuti Prostaglandins and Other Lipid Mediators, 2023 The COVID-19 pandemics has made sparkly evident the importance of acute inflammation and its timely resolution to protect humans from pathogenic viruses while sparing them from collateral damages due to an uncontrolled immune response. It is clear now that resolution of inflammation is an active process regulated by endogenous specialized proresolving lipid mediators (SPM) biosynthesized from essential polyunsaturated fatty acids. Accruing evidence indicates that SPM are produced during viral infections and play key roles in controlling the magnitude and duration of the inflammatory response and in regulating adaptive immunity. Here, we reviewed biosynthesis and bioactions of SPM in virus-mediated human diseases. Harnessing SPM and their proresolutive actions can help in providing new therapeutic approaches to current and future human viral diseases by controlling infection, stimulating host immunity, and protecting from organ damage.
Biomarkers of Response to Low-Dose Aspirin in Familial Adenomatous Polyposis Patients Angel Lanas, Stefania Tacconelli, Annalisa Contursi, Elena Piazuelo, Annalisa Bruno, Maurizio Ronci, Simone Marcone, Melania Dovizio, Federico Sopeña, Lorenza Falcone, Cristina Milillo, Matteo Mucci, Patrizia Ballerini, Paola Patrignani Cancers, 2023 Background: The results of Aspirin prevention of colorectal adenomas in patients with familial adenomatous polyposis (FAP) are controversial. Methods: We conducted a biomarker-based clinical study in eight FAP patients treated with enteric-coated low-dose Aspirin (100 mg daily for three months) to explore whether the drug targets mainly platelet cyclooxygenase (COX)-1 or affects extraplatelet cellular sources expressing COX-isozymes and/or off-target effects in colorectal adenomas. Results: In FAP patients, low-dose Aspirin-acetylated platelet COX-1 at Serine529 (>70%) was associated with an almost complete inhibition of platelet thromboxane (TX) B2 generation ex vivo (serum TXB2). However, enhanced residual urinary 11-dehydro-TXB2 and urinary PGEM, primary metabolites of TXA2 and prostaglandin (PG)E2, respectively, were detected in association with incomplete acetylation of COX-1 in normal colorectal biopsies and adenomas. Proteomics of adenomas showed that Aspirin significantly modulated only eight proteins. The upregulation of vimentin and downregulation of HBB (hemoglobin subunit beta) distinguished two groups with high vs. low residual 11-dehydro-TXB2 levels, possibly identifying the nonresponders and responders to Aspirin. Conclusions: Although low-dose Aspirin appropriately inhibited the platelet, persistently high systemic TXA2 and PGE2 biosynthesis were found, plausibly for a marginal inhibitory effect on prostanoid biosynthesis in the colorectum. Novel chemotherapeutic strategies in FAP can involve blocking the effects of TXA2 and PGE2 signaling with receptor antagonists.
Microvascular and Macrovascular Endothelial Cell Isolation and Purification from Lung-Derived Samples Roberto Plebani, Alessandra D'Alessandro, Paola Lanuti, Pasquale Simeone, Massimiliano Cinalli, Ilaria Righi, Alessandro Palleschi, Matteo Mucci, Marco Marchisio, Francesco Cappabianca, Marina Camera, Felice Mucilli, Mario Romano Journal of Visualized Experiments, 2023 The availability of cells isolated from healthy and diseased tissues and organs represents a key element for personalized medicine approaches. Although biobanks can provide a wide collection of primary and immortalized cells for biomedical research, these do not cover all experimental needs, particularly those related to specific diseases or genotypes. Vascular endothelial cells (ECs) are key components of the immune inflammatory reaction and, thus, play a central role in the pathogenesis of a variety of disorders. Notably, ECs from different sites display different biochemical and functional properties, making the availability of specific EC types (i.e., macrovascular, microvascular, arterial, and venous) essential for designing reliable experiments. Here, simple procedures to obtain high-yield, virtually pure human macrovascular and microvascular endothelial cells from the pulmonary artery and lung parenchyma are illustrated in detail. This methodology can be easily reproduced at a relatively low cost by any laboratory to achieve independence from commercial sources and obtain EC phenotypes/genotypes that are not yet available.
Maternal, newborn and breast milk concentrations of elexacaftor/tezacaftor/ivacaftor in a F508del heterozygous woman with cystic fibrosis following successful pregnancy Pietro Ripani, Matteo Mucci, Stefano Pantano, Maria Di Sabatino, Francesca Collini, Giulia Ferri, Mario Romano, Antonio Recchiuti Frontiers in Medicine, 2023 With the introduction of elexacaftor/tezacaftor/ivacaftor (ETI), more women with cystic fibrosis (CF) are likely to grow families. Hence, an understanding long-term safety and effects of CFTR modulators on fertile women and children while monitoring their concentrations is crucial. Here, we report on the development of an improved LC–MS/MS methodology to measure ETI concentrations in maternal and child blood and breastmilk, applied in one case of successful pregnancy of a 30-year-old woman with CF (F508del/R334W). We observed that ETI remains stable in breastmilk, is absorbed by the infant and can be detected in child plasma. Our results confirm accumulating evidence of a successful pregnancy in women treated with CFTR modulators without significant side effects on the child and provide valuable analytical procedures suitable for the post-marketing evaluation of CFTR modulators in pregnant and lactating women, as well as in their infants.
Characterization of the acetylation of cyclooxygenase-isozymes and targeted lipidomics of eicosanoids in serum and colon cancer cells by the new aspirin formulation IP1867B versus aspirin in vitro Ulrika Hofling, Stefania Tacconelli, Annalisa Contursi, Annalisa Bruno, Matteo Mucci, Patrizia Ballerini, Simon Cohen, Paola Patrignani Frontiers in Pharmacology, 2022 Background: Aspirin(acetylsalicylic acid, ASA) is recommended for the secondary prevention of atherothrombotic events and has shown anticancer effects. The current enteric-coated drug formulation may reduce aspirin bioavailability. Liquid formulations could improve aspirin pharmacokinetics and pharmacodynamics. IP1867B is a liquid-aspirin formulation that combines three ingredients, ASA/triacetin/saccharin.Methods: ASA and IP1867B(L-ASA) were assessed in human serum(obtained by allowing to clot human whole blood at 37 °C for 1h), washed platelets, and colonic adenocarcinoma HCA7 cells on eicosanoid generation and COX-isozyme acetylation at Serine529 and 516 by LC-MS/MS.Results: In serum, ASA and L-ASA acted by selectively affecting COX-1-derived eicosanoids, including thromboxane(TX)B2. L-ASA was more potent in inhibiting serum TXB2, a known biomarker of aspirin antiplatelet effect, than ASA. However, ASA and L-ASA were equipotent to acetylate COX-1 in washed platelets and COX-2 in HCA7 cells. In HCA7 cells, ASA and L-ASA acted by inhibiting prostaglandin(PG)E2(the most abundant prostanoid) and TXB2 biosynthesis. In the presence of a high arachidonic acid concentration(100 μM), 15R-hydroxyeicosatetraenoic acid(HETE) was generated at baseline by cancer cell COX-2 and was only slightly enhanced by supratherapeutic concentrations of ASA(1 mM). In whole blood and HCA7 cells treated with ASA or L-ASA, 15-epi-lipoxin(LX)A4 were undetectable.Conclusion: IP1867B was more potent in affecting serum TXB2 generation than ASA. The relevance of this finding deserves evaluation in vivo in humans. In cancer cells, ASA and IP1867B acted by inhibiting PGE2 and TXB2 generation via the acetylation of COX-2. ASA and IP867B at clinically relevant concentrations did not substantially induce the biosynthesis of 15R-HETE and 15-epi-LXA4.
