Genètica aplicada a la clínica neurovascular /

En aquesta tesi doctoral es descriuen els resultats de l'estudi farmacogenètic Geno-tPA, en el qual es va analitzar el paper de centenars de polimorfismes (SNPs) en la resposta al tractament trombolític de l'ictus isquèmic: recanalització arterial una hora després de la infusió, reoclusión arterial després d'haver recanalitzar, aparició de transformació hemorràgica iatrogènica i mortalitat intrahospitalària. Els resultats van ser confirmats en una altra cohort independent, sempre que la variable d'estudi pogués ser determinada. Es van generar models predictius amb les dades genètiques i clínics, que van ser validats en sèries clíniques independents. A més, es van realitzar estudis funcionals amb els polimorfismes associats per determinar el mecanisme mitjançant el qual s'associaven amb la variable clínica d'estudi. Aquests assaigs van incloure RT-qPCR, western blot, ELISA, nefelometria, proves de coagulació o arrays de proteïnes Searchlight. Finalment, es va seqüenciar la regió en desequilibri de lligament amb els millors marcadors per determinar si hi havia una altra variant funcional encara més associada al fenotip clínic que la inclosa en l'anàlisi inicial. Paral·lelament, es van implementar protocols de diagnòstic genètic per a les malalties monogèniques causants d'ictus CADASIL i CARASIL. Aquestes patologies es caracteritzen per presentar infarts llacunars repetitius d'inici tardà que condueixen a la demència vascular i la mort. Així, descrivim 2 noves mutacions causants de CADASIL a la nostra població, situades a l'exó 7 i 23 de NOTCH3, localitzacions infreqüents en l'espectre de la malaltia. Paral·lelament, descrivim el primer cas de CARASIL en població caucàsica per mutació a l'exó 4 del gen HTRA1, éssent recomanable la seva inclusió en el diagnòstic diferencial de les malalties de petit vas si el pacient presenta danys cerebrals, calvície primerenca i lesions a l'esquenaThis thesis describes the results of the Geno-tPA pharmacogenetic study, which analyzed the role of hundreds of polymorphisms (SNPs) in the response to thrombolytic treatmentof ischemic stroke: recanalization one hour after t-PA infusion, arterial reocclusion of the recanalized artery, presence of iatrogenic hemorrhagic transformation and in-hospital mortality. The results were confirmed in an independent cohort, provided the study variable could be determined. Predictive models generated with genetic and clinical data, which were validated in independent clinical series. In addition, we performed functional studies with the associated polymorphisms to determine the mechanism by which they are linked with the clinical variable. These trials included RT-qPCR, western blot, ELISA, nephelometry, coagulation or protein arrays Searchlight. Finally, we sequenced the region in linkage disequilibrium with the best SNP markers to determine whether there was another functional variant even more associated with clinical phenotype that those included in the initial analysis. In parallel, I implemented genetic testing protocols for monogenic diseases causing stroke (CARASIL and CADASIL). These diseases are characterized by repetitive lacunar infarcts leading to late-onset vascular dementia and death. Thus, we describe 2 new CADASIL-causing mutations in our population, located in exon7 and 23 of NOTCH3 gene, rare locations in the spectrum of the disease. In parallel, we describe the first case of CARASIL in Caucasian population by mutation in exon 4 of HTRA1 gene, being recommended its inclusion in the differential diagnosis of small vessel diseases if the patient suffers brain damage, early baldness and back injuries

Update on the Serum Biomarkers and Genetic Factors Associated with Safety and Efficacy of rt-PA Treatment in Acute Stroke Patients

An accurate understanding of the mechanisms underlying an individual's response to rt-PA treatment is critical to improve stroke patients' management. We thus reviewed the literature in order to identify biochemical and genetic factors that have been associated with safety and efficacy of rt-PA administration after stroke

Drug-induced chromatin accessibility changes associate with sensitivity to liver tumor promotion

Liver cancer susceptibility varies amongst humans and between experimental animal models due to multiple genetic and epigenetic factors. The molecular characterization of such susceptibilities has the potential to enhance cancer risk assessment of xenobiotic exposures and disease prevention strategies. Here, using DNase I hypersensitivity mapping coupled with transcriptomic profiling, we investigate perturbations in cis-acting gene regulatory elements associated with the early stages of phenobarbital (PB)- mediated liver tumor promotion in susceptible versus resistant mouse strains (B6C3F1 versus C57BL/6J). Integrated computational analyses of strain-selective changes in liver chromatin accessibility underlying PB-response reveal differential epigenetic regulation of molecular pathways associated with PB-mediated tumor promotion, including Wnt/-catenin signalling. Complementary transcription factor motif analyses reveal mouse strain-selective gene regulatory networks and a novel role for Stat, Smad and Fox transcription factors in the early stages of PB-mediated tumor promotion. Mapping perturbations in cis-acting gene regulatory elements provides novel insights into the molecular basis for susceptibility to xenobiotic-induced rodent liver tumor promotion and has the potential to enhance mechanism-based cancer risk assessments of xenobiotic exposures

Genètica aplicada a la clínica neurovascular

En aquesta tesi doctoral es descriuen els resultats de l'estudi farmacogenètic Geno-tPA, en el qual es va analitzar el paper de centenars de polimorfismes (SNPs) en la resposta al tractament trombolític de l'ictus isquèmic: recanalització arterial una hora després de la infusió, reoclusión arterial després d'haver recanalitzar, aparició de transformació hemorràgica iatrogènica i mortalitat intrahospitalària. Els resultats van ser confirmats en una altra cohort independent, sempre que la variable d'estudi pogués ser determinada. Es van generar models predictius amb les dades genètiques i clínics, que van ser validats en sèries clíniques independents. A més, es van realitzar estudis funcionals amb els polimorfismes associats per determinar el mecanisme mitjançant el qual s'associaven amb la variable clínica d'estudi. Aquests assaigs van incloure RT-qPCR, western blot, ELISA, nefelometria, proves de coagulació o arrays de proteïnes Searchlight. Finalment, es va seqüenciar la regió en desequilibri de lligament amb els millors marcadors per determinar si hi havia una altra variant funcional encara més associada al fenotip clínic que la inclosa en l'anàlisi inicial. Paral.lelament, es van implementar protocols de diagnòstic genètic per a les malalties monogèniques causants d'ictus CADASIL i CARASIL. Aquestes patologies es caracteritzen per presentar infarts llacunars repetitius d'inici tardà que condueixen a la demència vascular i la mort. Així, descrivim 2 noves mutacions causants de CADASIL a la nostra població, situades a l'exó 7 i 23 de NOTCH3, localitzacions infreqüents en l'espectre de la malaltia. Paral·lelament, describim el primer cas de CARASIL en població caucàsica per mutació a l’exó 4 del gen HTRA1, éssent recomanable la seva inclusió en el diagnòstic diferencial de les malalties de petit vas si el pacient presenta danys cerebrals, calvície primerenca i lesions a l’esquena.This thesis describes the results of the Geno-tPA pharmacogenetic study, which analyzed the role of hundreds of polymorphisms (SNPs) in the response to thrombolytic treatmentof ischemic stroke: recanalization one hour after t-PA infusion, arterial reocclusion of the recanalized artery, presence of iatrogenic hemorrhagic transformation and in-hospital mortality. The results were confirmed in an independent cohort, provided the study variable could be determined. Predictive models generated with genetic and clinical data, which were validated in independent clinical series. In addition, we performed functional studies with the associated polymorphisms to determine the mechanism by which they are linked with the clinical variable. These trials included RT-qPCR, western blot, ELISA, nephelometry, coagulation or protein arrays Searchlight. Finally, we sequenced the region in linkage disequilibrium with the best SNP markers to determine whether there was another functional variant even more associated with clinical phenotype that those included in the initial analysis. In parallel, I implemented genetic testing protocols for monogenic diseases causing stroke (CARASIL and CADASIL). These diseases are characterized by repetitive lacunar infarcts leading to late-onset vascular dementia and death. Thus, we describe 2 new CADASIL-causing mutations in our population, located in exon7 and 23 of NOTCH3 gene, rare locations in the spectrum of the disease. In parallel, we describe the first case of CARASIL in Caucasian population by mutation in exon 4 of HTRA1 gene, being recommended its inclusion in the differential diagnosis of small vessel diseases if the patient suffers brain damage, early baldness and back injuries

Update on the Serum Biomarkers and Genetic Factors Associated with Safety and Efficacy of rt-PA Treatment in Acute Stroke Patients

An accurate understanding of the mechanisms underlying an individual's response to rt-PA treatment is critical to improve stroke patients' management. We thus reviewed the literature in order to identify biochemical and genetic factors that have been associated with safety and efficacy of rt-PA administration after stroke

A predictive clinical-genetic model of tissue plasminogen activator response in acute ischemic stroke

Wide interindividual variability exists in response to tissue plasminogen activator (t-PA) treatment in the acute phase of ischemic stroke. We aimed to find genetic variations associated with hemorrhagic transformation (HT) and mortality rates after t-PA. We then generated a clinical-genetic model for predicting t-PA response

Efficacy and safety of the CVnCoV SARS-CoV-2 mRNA vaccine candidate in ten countries in Europe and Latin America (HERALD): a randomised, observer-blinded, placebo-controlled, phase 2b/3 trial

Background: Additional safe and efficacious vaccines are needed to control the COVID-19 pandemic. We aimed to analyse the efficacy and safety of the CVnCoV SARS-CoV-2 mRNA vaccine candidate. Methods: HERALD is a randomised, observer-blinded, placebo-controlled, phase 2b/3 clinical trial conducted in 47 centres in ten countries in Europe and Latin America. By use of an interactive web response system and stratification by country and age group (18–60 years and ≥61 years), adults with no history of virologically confirmed COVID-19 were randomly assigned (1:1) to receive intramuscularly either two 0·6 mL doses of CVnCoV containing 12 μg of mRNA or two 0·6 mL doses of 0·9% NaCl (placebo) on days 1 and 29. The primary efficacy endpoint was the occurrence of a first episode of virologically confirmed symptomatic COVID-19 of any severity and caused by any strain from 15 days after the second dose. For the primary endpoint, the trial was considered successful if the lower limit of the CI was greater than 30%. Key secondary endpoints were the occurrence of a first episode of virologically confirmed moderate-to-severe COVID-19, severe COVID-19, and COVID-19 of any severity by age group. Primary safety outcomes were solicited local and systemic adverse events within 7 days after each dose and unsolicited adverse events within 28 days after each dose in phase 2b participants, and serious adverse events and adverse events of special interest up to 1 year after the second dose in phase 2b and phase 3 participants. Here, we report data up to June 18, 2021. The study is registered at ClinicalTrials.gov, NCT04652102, and EudraCT, 2020–003998–22, and is ongoing. Findings: Between Dec 11, 2020, and April 12, 2021, 39 680 participants were enrolled and randomly assigned to receive either CVnCoV (n=19 846) or placebo (n=19 834), of whom 19 783 received at least one dose of CVnCoV and 19 746 received at least one dose of placebo. After a mean observation period of 48·2 days (SE 0·2), 83 cases of COVID-19 occurred in the CVnCoV group (n=12 851) in 1735·29 person-years and 145 cases occurred in the placebo group (n=12 211) in 1569·87 person-years, resulting in an overall vaccine efficacy against symptomatic COVID-19 of 48·2% (95·826% CI 31·0–61·4; p=0·016). Vaccine efficacy against moderate-to-severe COVID-19 was 70·7% (95% CI 42·5–86·1; CVnCoV 12 cases in 1735·29 person-years, placebo 37 cases in 1569·87 person-years). In participants aged 18–60 years, vaccine efficacy against symptomatic disease was 52·5% (95% CI 36·2–64·8; CVnCoV 71 cases in 1591·47 person-years, placebo, 136 cases in 1449·23 person-years). Too few cases occurred in participants aged 61 years or older (CVnCoV 12, placebo nine) to allow meaningful assessment of vaccine efficacy. Solicited adverse events, which were mostly systemic, were more common in CVnCoV recipients (1933 [96·5%] of 2003) than in placebo recipients (1344 [67·9%] of 1978), with 542 (27·1%) CVnCoV recipients and 61 (3·1%) placebo recipients reporting grade 3 solicited adverse events. The most frequently reported local reaction after any dose in the CVnCoV group was injection-site pain (1678 [83·6%] of 2007), with 22 grade 3 reactions, and the most frequently reported systematic reactions were fatigue (1603 [80·0%] of 2003) and headache (1541 [76·9%] of 2003). 82 (0·4%) of 19 783 CVnCoV recipients reported 100 serious adverse events and 66 (0·3%) of 19 746 placebo recipients reported 76 serious adverse events. Eight serious adverse events in five CVnCoV recipients and two serious adverse events in two placebo recipients were considered vaccination-related. None of the fatal serious adverse events reported (eight in the CVnCoV group and six in the placebo group) were considered to be related to study vaccination. Adverse events of special interest were reported for 38 (0·2%) participants in the CVnCoV group and 31 (0·2%) participants in the placebo group. These events were considered to be related to the trial vaccine for 14 (<0·1%) participants in the CVnCoV group and for five (<0·1%) participants in the placebo group. Interpretation: CVnCoV was efficacious in the prevention of COVID-19 of any severity and had an acceptable safety profile. Taking into account the changing environment, including the emergence of SARS-CoV-2 variants, and timelines for further development, the decision has been made to cease activities on the CVnCoV candidate and to focus efforts on the development of next-generation vaccine candidates. Funding: German Federal Ministry of Education and Research and CureVac