ISCIII - Artículoshttp://hdl.handle.net/20.500.12105/21032024-03-28T09:04:26Z2024-03-28T09:04:26ZA pilot study on the feasibility of European harmonized human biomonitoring: Strategies towards a common approach, challenges and opportunitiesCasteleyn, LDumez, BBecker, KKolossa-Gehring, MDen Hond, ESchoeters, GCastaño, ArgeliaKoch, H MAngerer, JEsteban-Lopez, MartaExley, KSepai, OBloemen, LHorvat, MKnudsen, L EJoas, AJoas, RBiot, PKoppen, GDewolf, M-CKatsonouri, AHadjipanayis, ACerná, MKrsková, ASchwedler, GFiddicke, UNielsen, J K SJensen, J FRudnai, PKözepésy, SMulcahy, MMannion, RGutleb, A CFischer, M ELigocka, DJakubowski, MReis, M FNamorado, SLupsa, I-RGurzau, A EHalzlova, KJajcaj, MMazej, DTratnik Snoj, JPosada De la Paz, ManuelLopez-Martin, EstrellaBerglund, MLarsson, KLehmann, ACrettaz, PAerts, Dhttp://hdl.handle.net/20.500.12105/175442024-02-08T12:01:18Z2015-08-01T00:00:00ZIn 2004 the European Commission and Member States initiated activities towards a harmonized approach for Human Biomonitoring surveys throughout Europe. The main objective was to sustain environmental health policy by building a coherent and sustainable framework and by increasing the comparability of data across countries. A pilot study to test common guidelines for setting up surveys was considered a key step in this process. Through a bottom-up approach that included all stakeholders, a joint study protocol was elaborated. From September 2011 till February 2012, 17 European countries collected data from 1844 mother-child pairs in the frame of DEMOnstration of a study to COordinate and Perform Human Biomonitoring on a European Scale (DEMOCOPHES).(1) Mercury in hair and urinary cadmium and cotinine were selected as biomarkers of exposure covered by sufficient analytical experience. Phthalate metabolites and Bisphenol A in urine were added to take into account increasing public and political awareness for emerging types of contaminants and to test less advanced markers/markers covered by less analytical experience. Extensive efforts towards chemo-analytical comparability were included. The pilot study showed that common approaches can be found in a context of considerable differences with respect to experience and expertize, socio-cultural background, economic situation and national priorities. It also evidenced that comparable Human Biomonitoring results can be obtained in such context. A European network was built, exchanging information, expertize and experiences, and providing training on all aspects of a survey. A key challenge was finding the right balance between a rigid structure allowing maximal comparability and a flexible approach increasing feasibility and capacity building. Next steps in European harmonization in Human Biomonitoring surveys include the establishment of a joint process for prioritization of substances to cover and biomarkers to develop, linking biomonitoring surveys with health examination surveys and with research, and coping with the diverse implementations of EU regulations and international guidelines with respect to ethics and privacy.
2015-08-01T00:00:00ZPlasma levels of α1-antitrypsin-derived C-terminal peptides in PiMM and PiZZ COPD patientsBörner, Friedemann RLechowicz, UrszulaWrenger, SabineMartinez-Delgado, BeatrizOlejnicka, BeataWelte, TobiasChorostowska-Wynimko, JoannaKiehntopf, MichaelJanciauskiene, Sabinahttp://hdl.handle.net/20.500.12105/173812024-02-01T02:01:52Z2023-11-01T00:00:00ZPlasma levels of α1-antitrypsin-derived C-terminal peptides might be valid as novel biomarkers to predict and/or characterise exacerbations in PiMM and PiZZ COPD patients, or to reflect the efficiency of augmentation therapy in PiZZ patients https://bit.ly/3rNJeLd.
2023-11-01T00:00:00ZImmunodetection of Truncated Forms of the α6 Subunit of the nAChR in the Brain of Spinosad Resistant Ceratitis capitata PhenotypesGuillem-Amat, AnaLópez-Errasquín, ElenaGarcía-Ricote, IreneBarbero, José LuisSánchez, LucasCasas-Tinto, SergioOrtego, Félixhttp://hdl.handle.net/20.500.12105/173802024-02-01T02:01:50Z2023-11-04T00:00:00ZThe α6 subunit of the nicotinic acetylcholine receptor (nAChR) has been proposed as the target for spinosad in insects. Point mutations that result in premature stop codons in the α6 gene of Ceratitis capitata flies have been previously associated with spinosad resistance, but it is unknown if these transcripts are translated and if so, what is the location of the putative truncated proteins. In this work, we produced a specific antibody against C. capitata α6 (Ccα6) and validated it by ELISA, Western blotting and immunofluorescence assays in brain tissues. The antibody detects both wild-type and truncated forms of Ccα6 in vivo, and the protein is located in the cell membrane of the brain of wild-type spinosad sensitive flies. On the contrary, the shortened transcripts present in resistant flies generate putative truncated proteins that, for the most part, fail to reach their final destination in the membrane of the cells and remain in the cytoplasm. The differences observed in the locations of wild-type and truncated α6 proteins are proposed to determine the susceptibility or resistance to spinosad.
2023-11-04T00:00:00ZEffects of maternal modafinil treatment on fetal development and neonatal growth parameters - a multicenter case series of the European Network of Teratology Information Services (ENTIS)Onken, MarliesLohse, LukasCoulm, BénédicteBeghin, DelphineRichardson, Jonathan LBermejo-Sanchez, EvaAguilera, CristinaBosch, MontserratCassina, MatteoChouchana, LaurentDe Santis, MarcoDuman, Mine KadiogluGören, M ZaferJohnson, DianaBera, Annie Pierre JonvilleKaplan, Yusuf CKennedy, DebraKwok, SusanLacroix, IsabelleLepelley, MarionPistelli, AlessandraSchaefer, ChristofTe Winkel, BernkeUysal, NusretWinterfeld, UrsulaYakuwa, NahoDiav-Citrin, OrnaVial, ThierryDathe, Katarinahttp://hdl.handle.net/20.500.12105/173792024-02-01T02:01:33Z2023-12-18T00:00:00ZObjective: In recent years, safety concerns about modafinil exposure during pregnancy have emerged. In particular, increased risks for major congenital anomalies (MCA) and impaired fetal growth were reported, although study results were conflicting. Our investigation aims to examine previously reported safety signals. Method: Multicenter case series based on data from 18 Teratology Information Services from 12 countries. Modafinil exposed pregnancies with an estimated date of birth before August 2019 were included in this study. For prospectively ascertained pregnancies, cumulative incidences of pregnancy outcomes, rate of nonchromosomal MCA in first trimester exposed pregnancies and percentiles of neonatal/infant weight and head circumference (HC) were calculated. Potential dose-dependent effects on fetal growth were explored by linear regression models. Retrospectively ascertained cases were screened for pattern of MCA and other adverse events. Results: One hundred and seventy-five prospectively ascertained cases were included, of which 173 were exposed at least during the first trimester. Cumulative incidences for live birth, spontaneous abortion and elective termination of pregnancy were 76.9% (95% CI, 68.0%-84.8%), 9.3% (95% CI, 5.0%-16.9%), and 13.9% (95% CI, 8.1%-23.1%), respectively. Nonchromosomal MCA was present in 3/150 live births, corresponding to an MCA rate of 2.0% (95%CI, 0.6%-6.1%), none were reported in pregnancy losses. Compared to reference standards, birth weight (BW) tended to be lower and neonatal HC to be smaller in exposed newborns (data available for 144 and 73 of 153 live births, respectively). In nonadjusted linear regression models, each 100 mg increase of average dosage per pregnancy day was associated with a decrease in standard deviation score (SDS) of -0.28 SDS (95% CI, -0.45 to -0.10) for BW and of -0.28 SDS (95% CI, -0.56 to 0.01) for HC. Screening of 22 retrospectively reported cases did not reveal any specific pattern of MCA or other adverse outcomes. Conclusion: The results do not indicate an increased risk of MCA after in utero exposure to modafinil, but a tendency toward lower BW and reduced neonatal HC. However, these findings should be regarded as preliminary. Until further studies allow for a definite conclusion, modafinil should not be used during pregnancy.
2023-12-18T00:00:00ZBronchoalveolar cytokine profile differentiates Pulmonary Langerhans cell histiocytosis patients from other smoking-related interstitial lung diseasesBarril, SilviaAcebo, PalomaMillan-Billi, PalomaLuque, AlfonsoSibila, OriolTarín, CarlosTazi, AbdellatifCastillo, DiegoHortelano, Sonsoleshttp://hdl.handle.net/20.500.12105/173772024-01-26T02:01:11Z2023-12-18T00:00:00ZBackground: Pulmonary Langerhans cell histiocytosis (PLCH) is a rare interstitial lung disease (ILD) associated with smoking, whose definitive diagnosis requires the exclusion of other forms of ILD and a compatible surgical lung biopsy. Bronchoalveolar lavage (BAL) is commonly proposed for the diagnosis of ILD, including PLCH, but the diagnostic value of this technique is limited. Here, we have analyzed the levels of a panel of cytokines and chemokines in BAL from PLCH patients, in order to identify a distinct immune profile to discriminate PLCH from other smoking related-ILD (SR-ILD), and comparing the results with idiopathic pulmonary fibrosis (IPF) as another disease in which smoking is considered a risk factor. Methods: BAL samples were collected from thirty-six patients with different ILD, including seven patients with PLCH, sixteen with SR-ILD and thirteen with IPF. Inflammatory profiles were analyzed using the Human Cytokine Membrane Antibody Array. Principal component analysis (PCA) was performed to reduce dimensionality and protein-protein interaction (PPI) network analysis using STRING 11.5 database were conducted. Finally, Random forest (RF) method was used to build a prediction model. Results: We have found significant differences (p < 0.05) on thirty-two cytokines/chemokines when comparing BAL from PLCH patients with at least one of the other ILD. Four main groups of similarly regulated cytokines were established, identifying distinct sets of markers for each cluster. Exploratory analysis using PCA (principal component analysis) showed clustering and separation of patients, with the two first components capturing 69.69% of the total variance. Levels of TARC/CCL17, leptin, oncostatin M (OSM) and IP-10/CXCL10 were associated with lung function parameters, showing positive correlation with FVC. Finally, random forest (RF) algorithm demonstrates that PLCH patients can be differentiated from the other ILDs based solely on inflammatory profile (accuracy 96.25%). Conclusions: Our results show that patients with PLCH exhibit a distinct BAL immune profile to SR-ILD and IPF. PCA analysis and RF model identify a specific immune profile useful for discriminating PLCH.
2023-12-18T00:00:00ZPhenolic and quinone methide nor-triterpenes as selective NLRP3 inflammasome inhibitorsGonzález-Cofrade, LauraGreen, Jack PCuadrado, IreneAmesty, ÁngelOramas-Royo, SandraBrough, DavidEstévez-Braun, AnaHortelano, Sonsolesde Las Heras, Beatrizhttp://hdl.handle.net/20.500.12105/173762024-01-26T02:01:26Z2023-03-01T00:00:00ZDysregulated inflammasome activity, particularly of the NLRP3 inflammasome, is associated with the development of several inflammatory diseases. The study of molecules directly targeting NLRP3 is an emerging field in the discovery of new therapeutic compounds for the treatment of inflammatory disorders. Friedelane triterpenes are biologically active phytochemicals having a wide range of activities including anti-inflammatory effects. In this work, we evaluated the potential anti-inflammatory activity of phenolic and quinonemethide nor-triterpenes (1-11) isolated from Maytenus retusa and some semisynthetic derivatives (12-16) through inhibition of the NLRP3 inflammasome in macrophages. Among them, we found that triterpenes 6 and 14 were the most potent, showing markedly reduced caspase-1 activity, IL-1β secretion (IC50 = 1.15 µM and 0.19 µM, respectively), and pyroptosis (IC50 = 2.21 µM and 0.13 µM, respectively). Further characterization confirmed their selective inhibition of NLRP3 inflammasome in both canonical and non-canonical activation pathways with no effects on AIM2 or NLRC4 inflammasome activation.
2023-03-01T00:00:00ZDehydrohispanolone Derivatives Attenuate the Inflammatory Response through the Modulation of Inflammasome ActivationGonzález-Cofrade, LauraOramas-Royo, SandraCuadrado, IreneAmesty, ÁngelHortelano, SonsolesEstevez-Braun, Anade Las Heras, Beatrizhttp://hdl.handle.net/20.500.12105/173752024-01-25T19:30:29Z2020-07-24T00:00:00ZThe NLRP3 inflammasome plays a critical role in inflammation-mediated human diseases and represents a promising drug target for novel anti-inflammatory therapies. Hispanolone is a labdane diterpenoid isolated from the aerial parts of Ballota species. This diterpenoid and some derivatives have demonstrated anti-inflammatory effects in classical inflammatory pathways. In the present study, a series of dehydrohispanolone derivatives (1-19) was synthesized, and their anti-inflammatory activities toward NLRP3 inflammasome activation were evaluated. The structures of the dehydrohispanolone analogues produced were elucidated by NMR spectroscopy and mass spectrometry. Four derivatives significantly inhibited IL-1β secretion, with 15 and 18 being the most active (IC50 = 18.7 and 13.8 μM, respectively). Analysis of IL-1β and caspase-1 expression revealed that the new diterpenoids 15 and 18 are selective inhibitors of the NLRP3 inflammasome, reinforcing the previously demonstrated anti-inflammatory properties of hispanolone derivatives.
2020-07-24T00:00:00ZCurrent status of terpenoids as inflammasome inhibitorsHortelano, SonsolesGonzález-Cofrade, LauraCuadrado, Irenede Las Heras, Beatrizhttp://hdl.handle.net/20.500.12105/173742024-01-26T02:01:31Z2020-02-01T00:00:00ZIncreasing evidence supports NLRP3 inflammasome as a new target to control inflammation. Dysregulation of NLRP3 inflammasome has been reported to be involved in the pathogenesis of several human inflammatory diseases. However, no NLRP3 inflammasome inhibitors are available in clinic. Terpenoids are natural products with multi-target activities against inflammation. Recent studies have revealed that these compounds are capable of inhibiting the activation of NLRP3 inflammasome in several mouse models of NLRP3 inflammasome-related pathogenesis. Thus, terpenoids represent an interesting pharmacological approach for the treatment of inflammatory diseases as they are endowed with a dual mechanism of inhibition of NF-KB transcription factor and inflammasome activation, both critically involved in their anti-inflammatory effects. This work provides an overview of the current knowledge on the therapeutic potential of terpenoids as NLRP3 inflammasome inhibitors.
2020-02-01T00:00:00ZA hispanolone-derived diterpenoid inhibits M2-Macrophage polarization in vitro via JAK/STAT and attenuates chitin induced inflammation in vivoJimenez-Garcia, LidiaHigueras, María ÁngelesHerranz, SandraHernández-López, MartaLuque, Alfonsode Las Heras, BeatrizHortelano, Sonsoleshttp://hdl.handle.net/20.500.12105/173722024-02-01T12:51:34Z2018-08-01T00:00:00ZMacrophages are highly plastic cells that adopt different functional phenotypes in response to environmental signals. Classically activated macrophages (M1) exhibit a pro-inflammatory role, mediating host defense against microorganisms or tumor cells; whereas alternatively activated macrophages (M2) perform a range of physiological processes, including inflammation, wound repair and tissue remodeling. Interestingly, M2 macrophages have been involved in pathological settings such as tumor progression, parasitic infection and respiratory disorders. Consequently, the search of new agents able to control macrophage polarization is on the basis of new therapeutic strategies. In the present study, we have evaluated the effect of the hispanolone derivative 8,9-dehydrohispanolone-15,16-lactol (DHHL) on M2 macrophage polarization. Our results reveal that DHHL significantly inhibited IL-4- or IL-13-stimulated M2 macrophage activation, as showed by reduced expression of M2 markers. In addition, DHHL suppressed IL-4-induced STAT-6 and JAK-1 tyrosine phosphorylation, suggesting that this compound inhibited M2 polarization by suppressing the JAK-STAT signaling pathway. Finally, DHHL prevented eosinophil recruitment and the presence of F4/80+-CD206+ M2-like macrophages in an in vivo model of M2 polarization via administration of chitin. Collectively, these results confirm DHHL as a novel regulator of macrophage polarization suitable to design future therapies towards M2-macrophages mediated pathologies.
2018-08-01T00:00:00ZMice inflammatory responses to inhaled aerosolized LPS: effects of various forms of human alpha1-antitrypsinSivaraman, KokilavaniWrenger, SabineLiu, BinSchaudien, DirkHesse, ChristinaGomez-Mariano, Gema MariaPerez-Luz, SaraSewald, KatherinaDeLuca, DavidWurm, Maria JPino, PacoWelte, TobiasMartinez-Delgado, BeatrizJanciauskiene, Sabinahttp://hdl.handle.net/20.500.12105/172692024-01-24T08:03:32Z2023-01-10T00:00:00ZRodent models of lipopolysaccharide (LPS)-induced pulmonary inflammation are used for anti-inflammatory drug testing. We aimed to characterize mice responses to aerosolized LPS alone or with intraperitoneal (i.p.) delivery of alpha1-antitrypsin (AAT). Balb/c mice were exposed to clean air or aerosolized LPS (0.21 mg/mL) for 10 min per day, for 3 d. One hour after each challenge, animals were treated i.p. with saline or with (4 mg/kg body weight) one of the AAT preparations: native (AAT), oxidized (oxAAT), recombinant (recAAT), or peptide of AAT (C-36). Experiments were terminated 6 h after the last dose of AATs. Transcriptome data of mice lungs exposed to clean air versus LPS revealed 656 differentially expressed genes and 155 significant gene ontology terms, including neutrophil migration and toll-like receptor signaling pathways. Concordantly, mice inhaling LPS showed higher bronchoalveolar lavage fluid neutrophil counts and levels of myeloperoxidase, inducible nitric oxide synthase, IL-1β, TNFα, KC, IL-6, and granulocyte-macrophage colony-stimulating factor (GM-CSF). Plasma inflammatory markers did not increase. After i.p. application of AATs, about 1% to 2% of proteins reached the lungs but, except for GM-CSF, none of the proteins significantly influenced inflammatory markers. All AATs and C-36 significantly inhibited LPS-induced GM-CSF release. Surprisingly, only oxAAT decreased the expression of several LPS-induced inflammatory genes, such as Cxcl3, Cd14, Il1b, Nfkb1, and Nfkb2, in lung tissues. According to lung transcriptome data, oxAAT mostly affected genes related to transcriptional regulation while native AAT or recAAT affected genes of inflammatory pathways. Hence, we present a feasible mice model of local lung inflammation induced via aerosolized LPS that can be useful for systemic drug testing.
2023-01-10T00:00:00ZDe novo small deletion affecting transcription start site of short isoform of AUTS2 gene in a patient with syndromic neurodevelopmental defectsMartinez-Delgado, BeatrizLopez-Martin, EstrellaLara-Herguedas, JuliánMonzon, SaraCuesta, IsabelJuliá, MiguelAquino, VirginiaRodriguez-Martin, CarlosDamian, AlejandraGonzalo, IreneGomez-Mariano, Gema MariaBaladron-Jimenez, Beatriz IsabelCazorla, RosarioIglesias, GemaRoman, EnriquetaRos, PurificacionTutor, PabloMellor, SusanaJimenez, CarlosCabrejas, Maria JoseGonzalez-Vioque, EmilianoAlonso, JavierBermejo-Sanchez, EvaPosada De la Paz, Manuelhttp://hdl.handle.net/20.500.12105/172682024-01-23T14:46:46Z2021-03-01T00:00:00ZDisruption of the autism susceptibility candidate 2 (AUTS2) gene through genomic rearrangements, copy number variations (CNVs), and intragenic deletions and mutations, has been recurrently involved in syndromic forms of developmental delay and intellectual disability, known as AUTS2 syndrome. The AUTS2 gene plays an important role in regulation of neuronal migration, and when altered, associates with a variable phenotype from severely to mildly affected patients. The more severe phenotypes significantly correlate with the presence of defects affecting the C-terminus part of the gene. This article reports a new patient with a syndromic neurodevelopmental disorder, who presents a deletion of 30 nucleotides in the exon 9 of the AUTS2 gene. Importantly, this deletion includes the transcription start site for the AUTS2 short transcript isoform, which has an important role in brain development. Gene expression analysis of AUTS2 full-length and short isoforms revealed that the deletion found in this patient causes a remarkable reduction in the expression level, not only of the short isoform, but also of the full AUTS2 transcripts. This report adds more evidence for the role of mutated AUTS2 short transcripts in the development of a severe phenotype in the AUTS2 syndrome.
2021-03-01T00:00:00ZmiR-320c Regulates SERPINA1 Expression and Is Induced in Patients With Pulmonary DiseaseMatamala, NereaLara, BeatrizGomez-Mariano, Gema MariaMartinez Rodríguez, SeleneVázquez-Domínguez, IreneOtero-Sobrino, ÁlvaroMuñoz-Callejas, AntonioSánchez, ElenaEsquinas, CristinaBustamante, AnaCadenas, SergioCuri, SergioLázaro, LourdesMartínez, María TeresaRodríguez, EstherMiravitlles, MarcTorres-Duran, MaríaHerrero, InésMichel, Francisco JavierCastillo, SilviaHernández-Pérez, José MªBlanco, IgnacioCasas, FranciscoMartinez-Delgado, Beatrizhttp://hdl.handle.net/20.500.12105/172672024-01-24T14:27:49Z2021-07-01T00:00:00ZIntroduction: Alpha-1 antitrypsin deficiency (AATD) is a genetic condition resulting in lung and liver disease with a great clinical variability. MicroRNAs have been identified as disease modifiers; therefore miRNA deregulation could play an important role in disease heterogeneity. Members of miR-320 family are involved in regulating of multiple processes including inflammation, and have potential specific binding sites in the 3'UTR region of SERPINA1 gene. In this study we explore the involvement of miR-320c, a member of this family, in this disease. Methods: Firstly in vitro studies were carried out to demonstrate regulation of SERPINA1 gene by miR-320. Furthermore, the expression of miR-320c was analyzed in the blood of 98 individuals with different AAT serum levels by using quantitative PCR and expression was correlated to clinical parameters of the patients. Finally, HL60 cells were used to analyze induction of miR-320c in inflammatory conditions. Results: Overexpression of miR-320 members in human HepG2 cells led to inhibition of SERPINA1 expression. Analysis of miR-320c expression in patient's samples revealed significantly increased expression of miR-320c in individuals with pulmonary disease. Additionally, HL60 cells treated with the pro-inflammatory factor lipopolysaccharide (LPS) showed increase in miR-320c expression, suggesting that miR-320c responds to inflammation. Conclusion: Our findings demonstrate that miR-320c inhibits SERPINA1 expression in a hepatic cell line and its levels in blood are associated with lung disease in a cohort of patients with different AAT serum levels. These results suggest that miR-320c can play a role in AAT regulation and could be a biomarker of inflammatory processes in pulmonary diseases.
2021-07-01T00:00:00ZGenome-wide profiling of non-smoking-related lung cancer cells reveals common RB1 rearrangements associated with histopathologic transformation in EGFR-mutant tumorsPros, ESaigi, MAlameda, DGomez-Mariano, Gema MariaMartinez-Delgado, BeatrizAlburquerque-Bejar, J JCarretero, JTonda, REsteve-Codina, ACatala, IPalmero, RJove, MLazaro, CPatiño-Garcia, AGil-Bazo, IVerdura, STeulé, ATorres-Lanzas, JSidransky, DReguart, NPio, RJuan-Vidal, ONadal, EFelip, EMontuenga, L MSanchez-Cespedes, Mhttp://hdl.handle.net/20.500.12105/172662024-01-24T14:21:10Z2020-02-01T00:00:00ZBackground: The etiology and the molecular basis of lung adenocarcinomas (LuADs) in nonsmokers are currently unknown. Furthermore, the scarcity of available primary cultures continues to hamper our biological understanding of non-smoking-related lung adenocarcinomas (NSK-LuADs). Patients and methods: We established patient-derived cancer cell (PDC) cultures from metastatic NSK-LuADs, including two pairs of matched EGFR-mutant PDCs before and after resistance to tyrosine kinase inhibitors (TKIs), and then performed whole-exome and RNA sequencing to delineate their genomic architecture. For validation, we analyzed independent cohorts of primary LuADs. Results: In addition to known non-smoker-associated alterations (e.g. RET, ALK, EGFR, and ERBB2), we discovered novel fusions and recurrently mutated genes, including ATF7IP, a regulator of gene expression, that was inactivated in 5% of primary LuAD cases. We also found germline mutations at dominant familiar-cancer genes, highlighting the importance of genetic predisposition in the origin of a subset of NSK-LuADs. Furthermore, there was an over-representation of inactivating alterations at RB1, mostly through complex intragenic rearrangements, in treatment-naive EGFR-mutant LuADs. Three EGFR-mutant and one EGFR-wild-type tumors acquired resistance to EGFR-TKIs and chemotherapy, respectively, and histology on re-biopsies revealed the development of small-cell lung cancer/squamous cell carcinoma (SCLC/LuSCC) transformation. These features were consistent with RB1 inactivation and acquired EGFR-T790M mutation or FGFR3-TACC3 fusion in EGFR-mutant tumors. Conclusions: We found recurrent alterations in LuADs that deserve further exploration. Our work also demonstrates that a subset of NSK-LuADs arises within cancer-predisposition syndromes. The preferential occurrence of RB1 inactivation, via complex rearrangements, found in EGFR-mutant tumors appears to favor SCLC/LuSCC transformation under growth-inhibition pressures. Thus RB1 inactivation may predict the risk of LuAD transformation to a more aggressive type of lung cancer, and may need to be considered as a part of the clinical management of NSK-LuADs patients.
2020-02-01T00:00:00ZNew cis-Acting Variants in PI*S Background Produce Null Phenotypes Causing Alpha-1 Antitrypsin DeficiencyMatamala, NereaGomez-Mariano, Gema MariaPerez, Jose AntonioBaladron-Jimenez, Beatriz IsabelTorres-Durán, MaríaMichel, Francisco JavierSaez, RaquelHernández-Pérez, Jose MaríaBelmonte, IreneRodriguez-Frias, FranciscoBlanco, IgnacioStrnad, PavelJanciauskiene, SabinaMartinez-Delgado, Beatrizhttp://hdl.handle.net/20.500.12105/172642024-01-24T08:02:59Z2020-10-01T00:00:00ZAlpha-1 antitrypsin deficiency (AATD) is an inherited condition characterized by reduced levels of serum AAT due to mutations in the SERPINA1 (Serpin family A member 1) gene. The Pi*S (Glu264Val) is one of the most frequent deficient alleles of AATD, showing high incidence in the Iberian Peninsula. Herein, we describe two new alleles carrying an S mutation but producing a null phenotype: QOVigo and QOAachen. The new alleles were identified by sequencing the SERPINA1 gene in three patients who had lower AAT serum levels than expected for the initial genotype. These alleles are the result of combined mutations in cis in a PI*S allele. Sequencing detected the S mutation in cis with Tyr138Cys (S+Tyr138Cys) in two patients, whereas a third one had the S mutation in cis with Pro391Thr variant (S+Pro391Thr). When expressed in a cellular model, these variants caused strong AAT polymerization and very low AAT secretion to almost undetectable levels. The isoelectric focusing method for plasma AAT phenotyping did not show AAT protein encoded by the novel mutant alleles, behaving as null. We called these alleles PI*S-plus because the S variant was phased with another variant conferring more aggressive characteristics to the allele. The current data demonstrate that the clinical variability observed in AATD can be explained by additional genetic variation, such as dual cis-acting variants in the SERPINA1 gene. The possible existence of other unrevealed variants combined in the PI*S alleles should be considered to improve the genetic diagnosis of the patients.
2020-10-01T00:00:00ZFamilial retinoblastoma due to intronic LINE-1 insertion causes aberrant and noncanonical mRNA splicing of the RB1 geneRodriguez-Martin, CarlosCidre, FlorenciaFernández-Teijeiro, AnaGomez-Mariano, Gema Mariade la Vega, LeticiaRamos, PatriciaZaballos, ÁngelMonzon-Fernandez, SaraAlonso, Javierhttp://hdl.handle.net/20.500.12105/172632024-01-24T07:59:59Z2016-05-01T00:00:00ZRetinoblastoma (RB, MIM 180200) is the paradigm of hereditary cancer. Individuals harboring a constitutional mutation in one allele of the RB1 gene have a high predisposition to develop RB. Here, we present the first case of familial RB caused by a de novo insertion of a full-length long interspersed element-1 (LINE-1) into intron 14 of the RB1 gene that caused a highly heterogeneous splicing pattern of RB1 mRNA. LINE-1 insertion was inferred by mRNA studies and full-length sequenced by massive parallel sequencing. Some of the aberrant mRNAs were produced by noncanonical acceptor splice sites, a new finding that up to date has not been described to occur upon LINE-1 retrotransposition. Our results clearly show that RNA-based strategies have the potential to detect disease-causing transposon insertions. It also confirms that the incorporation of new genetic approaches, such as massive parallel sequencing, contributes to characterize at the sequence level these unique and exceptional genetic alterations.
2016-05-01T00:00:00ZIdentification of genetic variants in pharmacokinetic genes associated with Ewing Sarcoma treatment outcomeRuiz-Pinto, SPita, GPatiño-García, AGarcía-Miguel, PAlonso, JavierPérez-Martínez, ASastre, AGomez-Mariano, Gema MariaLissat, AScotlandi, KSerra, MLadenstein, RLapouble, EPierron, GKontny, UPicci, PKovar, HDelattre, OGonzález-Neira, Ahttp://hdl.handle.net/20.500.12105/172622024-01-24T08:00:34Z2016-09-01T00:00:00ZBackground: Despite the effectiveness of current treatment protocols for Ewing sarcoma (ES), many patients still experience relapse, and survival following recurrence is <15%. We aimed to identify genetic variants that predict treatment outcome in children diagnosed with ES. Patients and methods: We carried out a pharmacogenetic study of 384 single-nucleotide polymorphisms (SNPs) in 24 key transport or metabolism genes relevant to drugs used to treat in pediatric patients (<30 years) with histologically confirmed ES. We studied the association of genotypes with tumor response and overall survival (OS) in a discovery cohort of 106 Spanish children, with replication in a second cohort of 389 pediatric patients from across Europe. Results: We identified associations with OS (P < 0.05) for three SNPs in the Spanish cohort that were replicated in the European cohort. The strongest association observed was with rs7190447, located in the ATP-binding cassette subfamily C member 6 (ABCC6) gene [discovery: hazard ratio (HR) = 14.30, 95% confidence interval (CI) = 1.53-134, P = 0.020; replication: HR = 9.28, 95% CI = 2.20-39.2, P = 0.0024] and its correlated SNP rs7192303, which was predicted to have a plausible regulatory function. We also replicated associations with rs4148737 in the ATP-binding cassette subfamily B member 1 (ABCB1) gene (discovery: HR = 2.96, 95% CI = 1.08-8.10, P = 0.034; replication: HR = 1.60, 95% CI = 1.05-2.44, P = 0.029), which we have previously found to be associated with poorer OS in pediatric osteosarcoma patients, and rs11188147 in cytochrome P450 family 2 subfamily C member 8 gene (CYP2C8) (discovery : HR = 2.49, 95% CI = 1.06-5.87, P = 0.037; replication: HR = 1.77, 95% CI = 1.06-2.96, P = 0.030), an enzyme involved in the oxidative metabolism of the ES chemotherapeutic agents cyclophosphamide and ifosfamide. None of the associations with tumor response were replicated. Conclusion: Using an integrated pathway-based approach, we identified polymorphisms in ABCC6, ABCB1 and CYP2C8 associated with OS. These associations were replicated in a large independent cohort, highlighting the importance of pharmacokinetic genes as prognostic markers in ES.
2016-09-01T00:00:00Z