Mesoporous CeO2 nanoparticles (MCNs) are promising for biomedical applications because of their intrinsic redox activity and favorable biocompatibility. In particular, MCNs with hollow and yolk-shell nanostructures have attracted considerable interest as nanocarriers due to their large internal volume, tunable shell porosity, and ability to regulate mass transport. However, the synthesis of hollow and yolk-shell MCNs remains challenging using conventional solution-based methods. Here, we report a versatile gas-phase spray pyrolysis approach for the synthesis of MCNs with tunable structures via atomization of cerium nitrate solutions in the presence of polyvinylpyrrolidone (PVP) as a structure-directing agent. By systematically varying precursor composition, droplet size, and thermal processing conditions, the internal cavity formation, particle morphology, and mesoporous structure were precisely controlled. A custom-designed multizone furnace further enabled stepwise observation of particle formation, providing mechanistic insights into the morphological evolution during spray pyrolysis. Moreover, surface functionalization of MCNs with thiolated dextran introduced redox-responsive behavior through disulfide linkages that are cleavable under intracellular reductive conditions. Biological evaluation demonstrated excellent biocompatibility in HEK293 cells and revealed that the Rhodamine 6G loading efficiency strongly correlates with internal void volume and particle morphology. Furthermore, the functionalized MCNs significantly enhanced the efficacy of the aminoglycoside antibiotic G418, promoting translational readthrough in U2OS reporter cells and restoring CFTR-R553X activity in epithelial monolayers. These findings highlight spray-pyrolyzed mesoporous CeO2 hollow and yolk-shell nanoparticles as promising redox-responsive nanocarriers for intracellular delivery and therapeutic modulation of genes affected by nonsense mutations.

The fabrication of thin macromolecular layers containing disulfide bonds, which can undergo reversible cleavage and reformation under redox conditions, holds great promise for advanced drug delivery applications. These layers enable rapid and reversible responses to specific biological stimuli. In this study, we introduce an approach utilizing a water-soluble, thiol-containing dextran as a polythiol grafted onto a thiol-functionalized flat substrate. Subsequently, a thiolated, dye-labeled dextran was immobilized onto the substrate via disulfide bonds under mild oxidative conditions, as verified by fluorescence imaging and quartz crystal microbalance analysis. The reversible release and reattachment of the dye in response to redox stimulation were successfully demonstrated. Furthermore, the designed redox-responsive dextran layer was applied to enhance the colloidal stability of cerium oxide (CeO2) nanoparticles, which inherently exhibit poor stability in aqueous media without a coating. This modification provided precise control over the particles stability in aqueous environments. Biocompatibility assessments using HEK293 cells confirmed that dextran-modified CeO2 nanoparticles maintained cell viability. These findings highlight the potential of redox-responsive coatings for tailoring colloidal stability and advancing nanotherapeutic applications, particularly in redox-sensitive microenvironments.

Background and Purpose Cystic fibrosis (CF) is due to loss-of-function variants of the CF transmembrane conductance regulator (CFTR) channel. The most effective treatment for people with CF carrying the F508del mutation is the triple combination of elexacaftor-tezacaftor-ivacaftor (ETI). ETI can correct the underlying defect(s) in other CFTR mutants. The use of disease-relevant predictive models such as patient-derived human nasal epithelial cells allow to investigate the response to CFTR modulators of specific genotypes, possibly supporting patients' access to treatment.Experimental Approach Using computational, biochemical and functional methodologies, a detailed analysis of selected variants in the intracellular loop 4 (ICL4) to understand their impact on CFTR structure and function.Key Results Mutations affecting L1065, R1066 and L1077 compromise structural stability of CFTR. Analyses of single variants expressed heterologously in immortalized bronchial cells showed that, upon ETI, rescued activity for both L1065P and R1066C was close to 50% of the wild-type CFTR activity. Biochemical studies of ICL4 variants expression pattern in CFBE41o-cells, following treatment for 24 h, demonstrate the appearance of the mature, fully glycosylated band, with no changes in the immature band. Finally, our study provides evidence in primary nasal cells from a cohort of people with CF that L1065P and R1066C can be effectively rescued by ETI up to 25%-45% of the activity measured in non-CF epithelia.Conclusion and Implications Although the observed rescue for L1065P and R1066C was smaller than that of the F508del, it should fall in a range predicted, by various studies, to provide a clinical benefit.

The Cystic Fibrosis Transmembrane conductance Regulator (CFTR) modulator VX-445 (Elexacaftor) used to treat cystic fibrosis presents both corrector and potentiator activities. This drug binds to a pocket within the CFTR membrane-spanning domain (MSD) assembly, in contact with the lasso motif. We have previously shown that the corrector activity of VX-445 is modulated by mutations within MSD1 and the nucleotide binding domain NBD1. Here, we evaluate if mutations affecting VX-445's corrector activity also affect its potentiator activity. Responses to increasing concentrations of VX-445 were measured using the halide sensitive fluorescent assay after transfection of CFTR mutants in HEK293 cells. Results show that VX-445 potentiated gating mutants causing cystic fibrosis located in the NBDs (NBD1 G551D and NBD2 G1349D) and in the IntraCellular Loops (ICL1 G178R and ICL3 G970R). Mutations within the VX-445 binding site inhibited potentiation of G551D, contrary to some mutations located outside this site that affected its corrector activity, two of which (M212A and F224A) were found to induce CFTR gain-of-function. Potentiation of G551D was also observed with correctors VX-809 and VX-121. In conclusion, CFTR modulator VX-445 can promote channel activity of gating mutants, an effect which is dependent on the integrity of its binding site. Potentiation could also be observed with correctors VX-809 and VX-121, indicating that more generally, CFTR correctors can promote channel activity.

Rationale:Respiratory status of people with Cystic Fibrosis (pwCF) carrying N1303K is improved by Elexacaftor/Tezacaftor/Ivacaftor (ETI) but, contrary to other mutations, the impact on sweat test results is limited. Methods:To explore this discrepancy, we implemented new sweat gland and respiratory cell lines stably expressing Wild type (WT)-, F508del- and N1303K-CFTR. CFTR dependent chloride (Cl-) and bicarbonate (HCO3-) transport was measured by short circuit current in these new models and in primary Human Nasal Epithelial Cells (HNECs). CFTR expression was evaluated by Western blot. Results:In the airway and the sweat gland cells expressing F508del-CFTR, ETI induced maturation of CFTR and increased Cl- transport. In the respiratory cell lines and HNECs, N1303K-CFTR generated both immature and mature forms of CFTR. Correction by ETI increased CFTR amounts without promoting its maturation and improved Cl- secretion. N1303K-CFTR channel activity was markedly increased by co-potentiation of IVA with Apigenin. In the sweat gland, N1303K-CFTR was expressed as a globally misfolded protein, non-rescuable by ETI. API treatment to 2 patients improved FEV1 without lowering sweat Cl- content. Conclusion:N1303K-CFTR shows tissue specific correction and suboptimal response to ETI which can be improved by API.

In the present investigation, redox-responsive-based dextran carriers were developed for the controlled release of hydrophobic molecules via a reducing agent naturally present in cells, namely glutathione. In this sense, dextran was modified with a thiol derivative. The roles of the hydrophilic segments in the molecular self-organisation of polysaccharide derivatives into nanoparticles were investigated by varying the average dextran molar mass. Crosslinked thiolated dextran particles were good carriers of hydrophobic molecules such as Nile red dye, which were efficiently encapsulated in the hydrophobic core and then selectively released. Indeed, the disulfide linkage connecting the hydrophobic tail with dextran was found to be cleaved by glutathione under physiological conditions for the fast release of Nile red. The cytotoxicity of the dextran-based particles was examined, and it was found to be nontoxic to living cells. Finally, we explored the versatility of dextran-derived particle drug carriers for pancreatic cancer treatment. The designed carriers were loaded with the anticancer drug paclitaxel and the obtained particles exhibited redox-responsive drug release and excellent anticancer activities with up to 90 % inhibition of cancer cell viability. Thus, the developed dextran derivatives are highly promising and suitable for a wide range of biomedical applications that require the loading and release of hydrophobic compounds.

The study aimed to create redox-responsive dextran carriers for controlled hydrophobic molecule release using glutathione, a natural cellular reducing agent, by modifying dextran with a thiol derivative. Investigating the impact of different hydrophobic length on the molecular self-organization of polysaccharide derivatives into nanoparticles helped to understand their roles in this process. The study demonstrated that thiolated dextran particles can be used as emulsifier and can effectively encapsulated hydrophobic molecules like Nile red dye, with the disulfide linkage being cleaved by glutathione under physiological conditions for rapid release. Additionally, the dextran-based particles were found to be non-toxic to living cells.

Nonsense mutations account for 12 % of cystic fibrosis (CF) cases. The presence of a premature termination codon (PTC) leads to gene inactivation, which can be countered by the use of drugs stimulating PTC readthrough, restoring production of the full-length protein. We recently identified a new readthrough inducer, TLN468, more efficient than gentamicin.We measured the readthrough induced by these two drugs with different cystic fibrosis transmembrane conductance regulator (CFTR) PTCs. We then determined the amino acids inserted at the S1196X, G542X, W846X and E1417X PTCs of CFTR during readthrough induced by gentamicin or TLN468. TLN468 significantly promoted the incorporation of one specific amino acid, whereas gentamicin did not greatly modify the proportions of the various amino acids incorporated relative to basal conditions. The function of the engineered missense CFTR channels corresponding to these four PTCs was assessed with and without potentiator. For the recoded CFTR, except for E1417Q and G542W, the PTC readthrough induced by TLN468 allowed the expression of CFTR variants that were correctly processed and had significant activity that was enhanced by CFTR modulators. These results suggest that it would be relevant to assess the therapeutic benefit of TLN468 PTC suppression in combination with CFTR modulators in preclinical assays.

This study presents a spray pyrolysis technique for fabricating mesoporous cerium oxide nanoparticles (MCNs) with tunable properties. The MCNs were extensively characterized by their morphology, crystallinity, optical properties, and surface area through transmission electron microscopy, scanning electron microscopy, X-ray diffraction, Raman spectroscopy, UV−vis spectroscopy, and BET adsorption isotherms. We manipulated synthesis parameters to tailor these characteristics, and subsequently evaluated the ability of MCNs to load and release the cationic probe Rhodamine 6G, demonstrating their responsiveness to pH variations. The particles showed high entrapment efficiency (up to 40 %), pH-responsive release, and pore-size-dependent release performance. Additionally, the biocompatibility of MCNs was confirmed by assessing their impact on the viability of HEK293 cells. The findings indicate that the fabricated MCNs hold promise for targeted nano-therapeutic applications, particularly within acidic microenvironments such as tumor sites.

Pulmonary arterial (PA) hypertension (PAH) is a severe cardiopulmonary disease that may be triggered by exposure to drugs such as dasatinib or facilitated by genetic predispositions. The incidence of dasatinib-associated PAH is estimated at 0.45%, suggesting individual predispositions. The mechanisms of dasatinib-associated PAH are still incomplete. We discovered a KCNK3 gene (Potassium channel subfamily K member 3; coding for outward K+ channel) variant in a patient with dasatinib-associated PAH and investigated the impact of this variant on KCNK3 function. Additionally, we assessed the effects of dasatinib exposure on KCNK3 expression. In control human PA smooth muscle cells (hPASMCs) and human pulmonary endothelial cells (hPECs), we evaluated the consequences of KCNK3 knockdown on cell migration, mitochondrial membrane potential, ATP production, and in vitro tube formation. Using mass spectrometry, we determined the KCNK3 interactome. Patch-clamp experiments revealed that the KCNK3 variant represents a loss-of-function variant. Dasatinib contributed to PA constriction by decreasing KCNK3 function and expression. In control hPASMCs, KCNK3 knockdown promotes mitochondrial membrane depolarization and glycolytic shift. Dasatinib exposure or KCNK3 knockdown reduced the number of caveolae in hPECs. Moreover, KCNK3 knockdown in control hPECs reduced migration, proliferation, and in vitro tubulogenesis. Using proximity labeling and mass spectrometry, we identified the KCNK3 interactome, revealing that KCNK3 interacts with various proteins across different cellular compartments. We identified a novel pathogenic variant in KCNK3 and showed that dasatinib downregulates KCNK3, emphasizing the relationship between dasatinib-associated PAH and KCNK3 dysfunction. We demonstrated that a loss of KCNK3-dependent signaling contributes to endothelial dysfunction in PAH and glycolytic switch of hPASMCs.

Avec le temps, la mucoviscidose est devenue un exemple de synergie entre la recherche en biologie cellulaire et les progrès cliniques. Les thérapies protéiques ont enfin apporté l’espoir d’une vie normale aux patients, bouleversant ainsi les statistiques épidémiologiques établies. Néanmoins, les patients ne guérissent pas, et l’évolution épidémiologique de la maladie ouvre de nouveaux défis pour la prise en charge des malades. Par ailleurs, environ 10 % des patients demeurent sans solution thérapeutique. De nouvelles stratégies sont ainsi envisagées et la communauté des chercheurs, industriels, patients et autorités de santé reste mobilisée pour suivre les effets à long terme de ces nouveaux traitements et explorer de nouvelles approches pharmacologiques.

Introduction N1303K is the fourth most frequent Cystic Fibrosis (CF) causing mutation. People with CF (pwCF) clinical status can be improved by Elexacaftor(ELX)/Tezacaftor(TEZ)/Ivacaftor (ETI) combotherapy. We investigated the mechanism underlying N1303K-CFTR rescue.Methods N1303K-CFTR expression and maturation was evaluated by Western Blot in cell lines and Human Nasal Epithelial Primary Cells (HNECs). Cell surface expression was studied by nanoluciferase complementation assay and TurboID proximity labeling. Functional rescue was tested in vitro by YFP-Based Assay and Short Circuit Current.Results Correction by ELX/TEZ increases N1303K-CFTR amounts, but not its maturation in CFTR-expressing HEK and 16HBEge cell lines and in HNECs. In control conditions, N1303K-CFTR is more distributed at the cell surface and significantly more surface partners are identified in the N1303K-CFTR interactome as compared to F508del-CFTR in HEK cells. ELX/TEZ induces a global stabilization of N1303K-CFTR without favoring its plasma membrane relocation in contrast to F508del-CFTR which is redistributed to the membrane. ETI increases N1303K-CFTR activity in HNECs and can be increased by API co-potentiation with a predicted increase in Forced Expiratory Volume in 1 second (ppFEV1) by respectively 13([2][1])% and 18%([3][2]). This is consistent with a gain in ppFEV1 reported in pwCF carrying the N1303K mutation and additional improvement by API in a patient.Conclusion These results support the expansion of ETI approval to N1303K mutation but highlight different mechanisms of action than for F508del.### Competing Interest StatementStefano Pantano declares Vertex pharmaceuticals support in sample collection without financial contribution. Stefano Costa declares payment or honoraria for speakers bureaus from Vertex pharmaceuticals. Sonia Volpi declares payment of honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from Vertex pharmaceuticals and DMF Pharma FoodAR and support for attending meetings and/or travel from Chiesi. Stephanie Bui declares participation in the protocoles of Vertex pharmaceuticals studies as principal investigator. Clemence Martin reports payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from Chiesi, Astra Zeneca, Boehringer Ingelheim, GSK. Support for attending meetings and/or travel from Chiesi, Boehringer Ingelheim. Nicoletta Pedemonte declares payment or honoraria from Vertex Pharmaceuticals for lectures, presentations, speakers bureaus, manuscript writing or educational events - Speaker for a lecture at the 45th European Cystic Fibrosis Society Conference, Rotterdam, June 2022. Pierre Regis Burgel reports grants from Vertex pharmaceuticals, GSK, outside the submitted work. Luis J.V. Galietta declares Patents planned, issued or pending. Compounds described are not present in the submitted paper. Isabelle Sermet-Gaudelus reports support for the present manuscript from Vaincre La Mucoviscidose and Mucoviscidose ABCF2. Isabelle Sermet-Gaudelus also reports, outside the submitted work, grants from Agence Nationale pour la Recherche, Assistance Publique Hopitaux de Paris, Vertex Innovation Award; consulting fees and travel support from Vertex therapeutics. [1]: #ref-2 [2]: #ref-3

CFTR is an anion channel that has evolved from the mold of an ABC transporter. It possesses specific structural features, including a lateral portal between the cytoplasmic extensions of its transmembrane helices TM4 and TM6. This TM4-TM6 portal is lined by basic residues attracting anions from the cytosol towards the intracellular vestibule. Even though a symmetric, open portal is not observed at the level of the TM10/TM12 interface, basic amino acids are also present at this level, exposed to solvent in the vicinity of the regulatory R region, whose phosphorylation enables channel activation. Here, using all-atom molecular dynamics simulations in combination with functional and biochemical assays, we investigate the importance of these basic amino acids (R1158 and R1030), and of a neighboring aromatic amino acid (W846) in the regulation of CFTR activity. Results indicate that mutation of these amino acids globally increased channel activity and enabled channel opening by potentiators without the need to elevate cAMP levels. These effects (i) were observed even when the binding site of the potentiator VX-770 was mutated, revealing a probable independent mechanism, and (ii) were additive to one gain-of-function mutant within the selectivity filter. Taken together, our results indicate that the region of the membrane-spanning domain 2 (MSD2), symmetric to the lateral portal located between MSD1 TM4 and TM6, is a novel critical actor of CFTR regulation.

Avec le temps, la mucoviscidose est devenue un exemple de synergie entre la recherche en biologie cellulaire et les progres cliniques. Les therapies proteiques ont enfin apporte l'espoir d'une vie normale aux patients, bouleversant ainsi les statistiques epidemiologiques etablies. Neanmoins, les patients ne guerissent pas, et l'evolution epidemiologique de la maladie ouvre de nouveaux defis pour la prise en charge des malades. Par ailleurs, environ 10 % des patients demeurent sans solution therapeutique. De nouvelles strategies sont ainsi envisagees et la communaute des chercheurs, industriels, patients et autorites de sante reste mobilisee pour suivre les effets a long terme de ces nouveaux traitements et explorer de nouvelles approches pharmacologiques.

The ATP-binding cassette (ABC) and solute carrier (SLC) transporters play pivotal roles in cellular transport mechanisms, influencing a wide range of physiological processes and impacting various medical conditions. Recent advancements in structural biology and computational modeling have provided significant insights into their function and regulation. This review provides an overview of the current knowledge of human ABC and SLC transporters, emphasizing their structural and functional relationships, transport mechanisms, and the contribution of computational approaches to their understanding. Current challenges and promising future research and methodological directions are also discussed.

We have previously shown that the CBb subunit of crotoxin, a β-neurotoxin with phospholipase A2 (PLA2) activity, targets the human ΔF508CFTR chloride channel implicated in cystic fibrosis (CF). By direct binding to the nucleotide binding domain 1 (NBD1) of ΔF508CFTR, this neurotoxic PLA2 acts as a potentiator increasing chloride channel current and corrects the trafficking defect of misfolded ΔF508CFTR inside the cell. Here, for a therapeutics development of new anti-cystic fibrosis agents, we use a structure-based in silico approach to design peptides mimicking the CBb-ΔF508NBD1 interface. Combining biophysical and electrophysiological methods, we identify several peptides that interact with the ΔF508NBD1 domain and reveal their effects as potentiators on phosphorylated ΔF508CFTR. Moreover, protein-peptide interactions and electrophysiological studies allowed us to identify key residues of ΔF508NBD1 governing the interactions with the novel potentiators. The designed peptides bind to the same region as CBb phospholipase A2 on ΔF508NBD1 and potentiate chloride channel activity. Certain peptides also show an additive effect towards the clinically approved VX-770 potentiator. The identified CF therapeutics peptides represent a novel class of CFTR potentiators and illustrate a strategy leading to reproducing the effect of specific protein-protein interactions.

BACKGROUND:Around 20% of people with cystic fibrosis (pwCF) do not have access to the triple combination elexacaftor/tezacaftor/ivacaftor (ETI) in Europe because they do not carry the F508del allele on the CF transmembrane conductance regulator (CFTR) gene. Considering that pwCF carrying rare variants may benefit from ETI, including variants already validated by the US Food and Drug Administration (FDA), a compassionate use programme was launched in France. PwCF were invited to undergo a nasal brushing to investigate whether the pharmacological rescue of CFTR activity by ETI in human nasal epithelial cell (HNEC) cultures was predictive of the clinical response. METHODS:CFTR activity correction was studied by short-circuit current in HNEC cultures at basal state (dimethyl sulfoxide (DMSO)) and after ETI incubation and expressed as percentage of normal (wild-type (WT)) CFTR activity after sequential addition of forskolin and Inh-172 (ΔI ETI/DMSO%WT). RESULTS:11 pwCF carried variants eligible for ETI according to the FDA label and 28 carried variants not listed by the FDA. ETI significantly increased CFTR activity of FDA-approved CFTR variants (I601F, G85E, S492F, M1101K, R347P, R74W;V201M;D1270N and H1085R). We point out ETI correction of non-FDA-approved variants, including N1303K, R334W, R1066C, Q552P and terminal splicing variants (4374+1G>A and 4096-3C>G). ΔI ETI/DMSO%WT was significantly correlated to change in percentage predicted forced expiratory volume in 1 s and sweat chloride concentration (p<0.0001 for both). G85E, R74W;V201M;D1270N, Q552P and M1101K were rescued more efficiently by other CFTR modulator combinations than ETI. CONCLUSIONS:Primary nasal epithelial cells hold promise for expanding the prescription of CFTR modulators in pwCF carrying rare mutants. Additional variants should be discussed for ETI indication.

Cystic fibrosis (CF) is caused by defective Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein. CFTR controls chloride (Cl-) and bicarbonate (HCO3-) transport into the Airway Surface Liquid (ASL).We investigated the impact of F508del-CFTR correction on HCO3- secretion by studying transepithelial HCO3- fluxes.HCO3- secretion was measured by pH-stat techniquein primary human respiratory epithelial cells from healthy subjects (WT) and people with CF (pwCF)carrying at least oneF508del variant.Its changes after CFTR modulation by the triple combination VX445/661/770 and in the context of TNF-α+IL-17 induced inflammation were related to ASL pH and transcriptionnal levels of CFTRand other HCO3- transporters ofairway epithelia such asSLC26A4 (Pendrin), SLC26A9 and NBCe1.CFTR-mediated HCO3-secretion was not detected in F508del primary human respiratory epithelial cells. It was rescued up to ∼ 80% of the WT levelby VX-445/661/770. In contrast,TNF-α+IL-17 normalized transepithelial HCO3-transportand ASL acidic pH. This was related to anincrease in SLC26A4 and CFTR transcript levels.VX-445/661/770 induced an increase in pH only in the context of inflammation.Effects on HCO3- transport werenot differentbetween F508del homozygous and F508del heterozygous CF airway epithelia.Our studies show that correction of F508del-CFTRHCO3- is not sufficient to buffer acidic ASL and that inflammation is a key regulator of HCO3-secretion in CF airways. Prediction of the response to CFTR modulators by theratyping should take into account airway inflammation.### Competing Interest StatementI Sermet-Gaudelus (ISG) is principal investigator of Vertex initiated studies ISG, Luis Galietta, Gilles Crambert and Gabrielle Planelles declare academic grant funded by Vertex Innovation Award. ### Funding StatementThis research was funded by Vertex Innovation Award-2017 cycle. Association ABCF, Vaincre La Mucoviscidose - RC20200502648-2020 Award, Miroslaw Zajac was financed by the Polish National Agency for Academic Exchange within Bekker Program no. BPN/BEK/2021/1/00284 and by National Science Center (NCN) Poland no. 2019/35/B/NZ1/02546.### Author DeclarationsI confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.YesThe details of the IRB/oversight body that provided approval or exemption for the research described are given below:The study was approved by the Ile de France 2 Ethics Committee, and written, informed consent was obtained from each adult and from both parents for participants below 18 (AFSSAPS (ANSM) B1005423-40, Eudract 2010-A00392-37; CPP IDF2: 2010-05-03-3).I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals.YesI understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance).YesI have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable.YesAll data produced in the present study are available upon reasonable request to the authors