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1. Ultra Deep Sequencing of Circulating Cell-Free DNA as a Potential Tool for Hepatocellular Carcinoma Management

Author

Mónica Higuera, Elena Vargas-Accarino, María Torrens, Josep Gregori, María Teresa Salcedo, Joan Martínez-Campreciós, Gloria Torres, María Bermúdez-Ramos, Itxarone Bilbao, Mercedes Guerrero-Murillo, Xavier Serres-Créixams, Xavier Merino, Francisco Rodríguez-Frías, Josep Quer, Beatriz Mínguez, Institut Català de la Salut, [Higuera M, Torrens M, Torres G] Grup de Recerca en Malalties Hepàtiques, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. [Vargas-Accarino E, Martínez-Campreciós J] Grup de Recerca en Malalties Hepàtiques, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Departament de Medicina, Universitat Autònoma de Barcelona, Bellaterra, Spain. [Gregori J] Grup de Recerca en Hepatitis Viral, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain. [Salcedo MT] Departament de Medicina, Universitat Autònoma de Barcelona, Bellaterra, Spain. Servei d’Anatomia Patològica, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Spanish Biomedical Research Network Centre in Oncology (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain. [Bermúdez-Ramos M] Grup de Recerca en Malalties Hepàtiques, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Departament de Medicina, Universitat Autònoma de Barcelona, Bellaterra, Spain. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain. [Bilbao I] Departament de Medicina, Universitat Autònoma de Barcelona, Bellaterra, Spain. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain. Servei de Cirurgia Hepatobiliopancreàtica i Trasplantaments, Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Guerrero-Murillo M] Grup de Recerca en Hepatitis Viral, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. [Serres-Créixams X, Merino X] Servei de Radiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Rodríguez-Frías F] Servei de Bioquímica, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Servei de Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, Bellaterra, Spain. [Quer J] Grup de Recerca en Hepatitis Viral, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain. Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, Bellaterra, Spain. [Mínguez B] Grup de Recerca en Malalties Hepàtiques, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Departament de Medicina, Universitat Autònoma de Barcelona, Bellaterra, Spain. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Instituto de Salud Carlos III, Madrid, Spain. Unitat Hepàtica, Vall d’Hebron Hospital Universitari, Barcelona, Spain, and Vall d'Hebron Barcelona Hospital Campus

Subjects
Cancer Research, técnicas de investigación::técnicas genéticas::análisis de secuencias::análisis de secuencias de ADN [TÉCNICAS Y EQUIPOS ANALÍTICOS, DIAGNÓSTICOS Y TERAPÉUTICOS], liquid biopsy, cfDNA, HCC, biomarkers, neoplasias::neoplasias::neoplasias por localización::neoplasias del sistema digestivo::neoplasias hepáticas::carcinoma hepatocelular [ENFERMEDADES], Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::Cell-Free Nucleic Acids [CHEMICALS AND DRUGS], Other subheadings::/therapy [Other subheadings], Fetge - Càncer - Aspectes genètics, Investigative Techniques::Genetic Techniques::Sequence Analysis::Sequence Analysis, DNA [ANALYTICAL, DIAGNOSTIC AND THERAPEUTIC TECHNIQUES, AND EQUIPMENT], Fetge - Càncer - Tractament, Àcids nucleics - Anàlisi, Neoplasms::Neoplasms::Neoplasms by Site::Digestive System Neoplasms::Liver Neoplasms::Carcinoma, Hepatocellular [DISEASES], nucleótidos y nucleósidos de ácidos nucleicos::ácidos nucleicos::ácidos nucleicos libres de células [COMPUESTOS QUÍMICOS Y DROGAS], Oncology, Otros calificadores::/terapia [Otros calificadores]
Abstract

Biomarkers; cfDNA; Liquid biopsy Biomarcadores; cfDNA; Biopsia líquida Biomarcadors; cfDNA; Biòpsia líquida Background: Cell-free DNA (cfDNA) concentrations have been described to be inversely correlated with prognosis in cancer. Mutations in HCC-associated driver genes in cfDNA have been reported, but their relation with patient’s outcome has not been described. Our aim was to elucidate whether mutations found in cfDNA could be representative from those present in HCC tissue, providing the rationale to use the cfDNA to monitor HCC. Methods: Tumoral tissue, paired nontumor adjacent tissue and blood samples were collected from 30 HCC patients undergoing curative therapies. Deep sequencing targeting HCC driver genes was performed. Results: Patients with more than 2 ng/µL of cfDNA at diagnosis had higher mortality (mean OS 24.6 vs. 31.87 months, p = 0.01) (AUC = 0.782). Subjects who died during follow-up, had a significantly higher number of mutated genes (p = 0.015) and number of mutations (p = 0.015) on cfDNA. Number of mutated genes (p = 0.001), detected mutations (p = 0.001) in cfDNA and ratio (number of mutations/cfDNA) (p = 0.003) were significantly associated with recurrence. However, patients with a ratio (number of mutations/cfDNA) above 6 (long-rank p = 0.0003) presented a higher risk of recurrence than those with a ratio under 6. Detection of more than four mutations in cfDNA correlated with higher risk of death (long-rank p = 0.042). Conclusions: In summary, cfDNA and detection of prevalent HCC mutations could have prognostic implications in early-stage HCC patients This research was funded by Instituto de Salud Carlos III (ISCIII) (PI18/00961) and (PI21/00714) to B.M. PI19/00301 by Instituto de Salud Carlos III (ISCIII) and Centro para el Desarrollo Tecnológico Industrial (CDTI) from the Spanish Ministry of Economy and Business, grant number IDI-20200297 to J.Q., E.V.-A. is a recipient of a predoctoral fellowship from Instituto de Salud Carlos III (ISCIII) (FI18/00027).

Published
2022
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2. Microorganisms as Shapers of Human Civilization, from Pandemics to Even Our Genomes: Villains or Friends? A Historical Approach

Author

Francisco Rodríguez-Frías, Josep Quer, David Tabernero, Maria Francesca Cortese, Selene Garcia-Garcia, Ariadna Rando-Segura, Tomas Pumarola, Institut Català de la Salut, [Rodríguez-Frías F] Grup de Recerca en Bioquímica Clínica, Servei de Bioquímica, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. Universitat Autònoma de Barcelona, Bellaterra, Spain. Unitat d’Hepatologies, Servei de Bioquímica i Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Universitat Autònoma de Barcelona, Bellaterra, Spain. [Quer J] Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, Instituto de Salud Carlos III, 28029 Madrid, Spain. Unitat del Fetge, Laboratori de Malalties Hepàtiques, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Tabernero D] Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, Instituto de Salud Carlos III, 28029 Madrid, Spain. [Cortese MF, Garcia-Garcia S] Grup de Recerca en Bioquímica Clínica, Servei de Bioquímica, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. Universitat Autònoma de Barcelona, Bellaterra, Spain. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, Instituto de Salud Carlos III, 28029 Madrid, Spain. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, Instituto de Salud Carlos III, 28029 Madrid, Spain. [Rando-Segura A] Unitat d’Hepatologies, Servei de Bioquímica i Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Universitat Autònoma de Barcelona, Bellaterra, Spain. Servei de Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Pumarola T] Servei de Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain, and Vall d'Hebron Barcelona Hospital Campus

Subjects
Microbiology (medical), humanidades::historia [HUMANIDADES], Yersinia pestis, QH301-705.5, Microorganismes, Genetic Phenomena::Genetic Structures::Genome::Genome, Microbial::Genome, Bacterial [PHENOMENA AND PROCESSES], fenómenos genéticos::estructuras genéticas::genoma::genoma microbiano::genoma bacteriano [FENÓMENOS Y PROCESOS], Review, pandemics, Microbiology, endogenous retrovirus, Virology, Plagues - Història, microbiota, Biology (General), SARS-CoV-2, virus diseases, population genetics, COVID-19, infecciones bacterianas y micosis::infecciones bacterianas::infecciones por bacterias gramnegativas::infecciones por Enterobacteriaceae::yersiniosis::peste [ENFERMEDADES], Genomes bacterians, Bacterial Infections and Mycoses::Bacterial Infections::Gram-Negative Bacterial Infections::Enterobacteriaceae Infections::Yersinia Infections::Plague [DISEASES], plague, Humanities::History [HUMANITIES], influenza, biotechnology
Abstract

SARS-CoV-2; Influenza; Microbiota SARS-CoV-2; Influenza; Microbiota SARS-CoV-2; Influenza; Microbiota Universal history is characterized by continuous evolution, in which civilizations are born and die. This evolution is associated with multiple factors, among which the role of microorganisms is often overlooked. Viruses and bacteria have written or decisively contributed to terrible episodes of history, such as the Black Death in 14th century Europe, the annihilation of pre-Columbian American civilizations, and pandemics such as the 1918 Spanish flu or the current COVID-19 pandemic caused by the coronavirus SARS-CoV-2. Nevertheless, it is clear that we could not live in a world without these tiny beings. Endogenous retroviruses have been key to our evolution and for the regulation of gene expression, and the gut microbiota helps us digest compounds that we could not otherwise process. In addition, we have used microorganisms to preserve or prepare food for millennia and more recently to obtain drugs such as antibiotics or to develop recombinant DNA technologies. Due to the enormous importance of microorganisms for our survival, they have significantly influenced the population genetics of different human groups. This paper will review the role of microorganisms as “villains” who have been responsible for tremendous mortality throughout history but also as “friends” who help us survive and evolve.

Published
2021
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3. Utility of the Cobas® Plasma Separation Card as a Sample Collection Device for Serological and Virological Diagnosis of Hepatitis C Virus Infection

Author

Fernando Velásquez-Orozco, Paula Salmerón, Josep Quer, Joan Martínez-Campreciós, Adrián Najarro-Centeno, Francisco Rodriguez-Frias, Maria Buti, Àngels Esteban, Ariadna Rando-Segura, Tomas Pumarola-Sune, Institut Català de la Salut, [Velásquez-Orozco F, Salmeron P, Pumarola-Suñe T] Servei de Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Rando-Segura A] Servei de Microbiologia, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, Spain. Servei d’Hepatologia. Servei de Microbiologia i Bioquímica, Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Martínez-Camprecios J] Servei de Medicina Interna, Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Najarro-Centeno A, Esteban À] Servei d’Hepatologia. Servei de Microbiologia i Bioquímica, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Unitat del Fetge, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. Servei de Bioquímica, Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Quer J] Unitat del Fetge, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. [Buti M] Servei de Medicina Interna, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Unitat del Fetge, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. CIBER de Enfermedades Hepáticas y Digestivas (CIBEREHD), Instituto de Salud Carlos III, Madrid, Spain. [Rodríguez-Frías F] Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, Spain. Servei d’Hepatologia. Servei de Microbiologia i Bioquímica, Vall d’Hebron Hospital Universitari, Barcelona, Spain. Unitat del Fetge, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Vall d’Hebron Hospital Universitari, Barcelona, Spain. Servei de Bioquímica, Vall d’Hebron Hospital Universitari, Barcelona, Spain. CIBER de Enfermedades Hepáticas y Digestivas (CIBEREHD), Instituto de Salud Carlos III, Madrid, Spain, and Vall d'Hebron Barcelona Hospital Campus

Subjects
Virus de l'hepatitis C - Diagnòstic, 0301 basic medicine, Plasma separation card, Hepatitis C virus, 030106 microbiology, Clinical Biochemistry, Otros calificadores::/diagnóstico [Otros calificadores], medicine.disease_cause, Article, virosis::virosis::infecciones por virus ARN::infecciones por Flaviviridae::hepatitis C [ENFERMEDADES], Serology, 03 medical and health sciences, 0302 clinical medicine, Other subheadings::/diagnosis [Other subheadings], dried plasma spot, medicine, Otros calificadores::/instrumentación [Otros calificadores], lcsh:R5-920, medicine.diagnostic_test, business.industry, Sang - Anàlisi, Antibody titer, Virus Diseases::Virus Diseases::RNA Virus Infections::Flaviviridae Infections::Hepatitis C [DISEASES], Hepatitis C, medicine.disease, Virology, Other subheadings::/instrumentation [Other subheadings], Titer, diagnóstico::técnicas y procedimientos diagnósticos::técnicas de laboratorio clínico::pruebas de bioquímica clínica::análisis químico de la sangre::análisis de manchas de sangre seca [TÉCNICAS Y EQUIPOS ANALÍTICOS, DIAGNÓSTICOS Y TERAPÉUTICOS], Immunoassay, 030211 gastroenterology & hepatology, Sample collection, Dried plasma spot, hepatitis C, lcsh:Medicine (General), business, Diagnosis::Diagnostic Techniques and Procedures::Clinical Laboratory Techniques::Clinical Chemistry Tests::Blood Chemical Analysis::Dried Blood Spot Testing [ANALYTICAL, DIAGNOSTIC AND THERAPEUTIC TECHNIQUES, AND EQUIPMENT], plasma separation card, Viral load
Abstract

Taca de plasma sec; Hepatitis C; Targeta de separació de plasma Mancha de plasma seca; Hepatitis C; Tarjeta de separación de plasma Dried plasma spot; Hepatitis C; Plasma separation card Diagnosis and clinical management of people infected with hepatitis C virus (HCV) relies on results from a combination of serological and virological tests. The aim of this study was to compare the performance of dried plasma spots (DPS), prepared using the cobas® Plasma Separation Card (PSC), to plasma and serum from venipuncture, for HCV diagnosis. We carried out a prospective study using DPS and paired plasma or serum samples. Serum and DPS samples were analyzed by immunoassay using Elecsys® Anti-HCV II (Roche). Plasma and DPS samples were analyzed using the cobas® HCV viral load and cobas® HCV genotyping tests (Roche). All DPS samples that had high anti-HCV antibody titers in serum were also antibody-positive, as were five of eight samples with moderate titers. Eight samples with low titers in serum were negative with DPS. Among 80 samples with plasma HCV viral loads between 61.5 and 2.2 × 108 IU/mL, 74 were RNA-positive in DPS. The mean viral load difference between plasma and DPS was 2.65 log10 IU/mL. The performance of DPS for detection of serological and virological markers of hepatitis C virus infection was comparable to that of the conventional specimen types. However, the limits of detection were higher for DPS. This work was supported by Roche and a microelimiation grant of Gilead Science (GLD19-0104).

Published
2021
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4. Prevalence and Risk Factors of MASLD and Liver Fibrosis amongst the Penitentiary Population in Catalonia: The PRISONAFLD Study.

Author

Rivera-Esteban J, Jiménez-Masip A, Muñoz-Martínez S, Augustin S, Guerrero RA, Gabriel-Medina P, Ferrer-Costa R, Rodríguez-Frías F, Turu E, Marco A, Pericàs JM, and On Behalf Of The Prisonafld Study Group Collaborators

Abstract

Background and Aims: The prevalence of chronic non-communicable diseases, particularly metabolic syndrome (MetS), has increased among the prison population. Nevertheless, we have limited data on metabolic dysfunction-associated steatotic liver disease (MASLD), the hepatic manifestation of this syndrome. We aimed to investigate the prevalence and risk factors of MASLD and MASLD-associated liver fibrosis in the penitentiary population in Catalonia, Spain., Method: A cross-sectional observational study involving eight penitentiary centers. Participants had at least one metabolic disorder and were at a closed-regimen penitentiary. Individuals with concomitant liver diseases and/or alcohol risk consumption were excluded. Significant fibrosis and MASLD were defined as liver stiffness ≥8 kPa and a controlled attenuation parameter ≥275 dB/m by vibration-controlled transient elastography (VCTE), respectively. After exclusions, metabolic inmates with VCTE were analyzed. Logistic regression analysis was performed to identify predictors of MASLD and MASLD-associated significant fibrosis., Results: Out of the 4338 inmates studied, 1290 (29.7%) had metabolic disorders, and 646 (14.9%) underwent VCTE. The mean age was 48.0 years (SD 12.1), and 89.5% were male. MASLD prevalence was 33.9%. Significant fibrosis and MASLD-associated significant fibrosis were found in 16.4% and 9.4% of inmates, respectively. In the multivariate analysis, T2D, waist circumference, MetS, and higher ALT values were identified as independent risk factors for MASLD and MASLD-associated significant fibrosis amongst the prison population., Conclusions: Metabolic disorders including MASLD are highly prevalent among inmates. The prevalence of significant fibrosis seems notably higher than that of the general population, underscoring the need for targeted screening programs and therapeutic interventions in the incarcerated population.

Published
2023
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5. Soluble Angiotensin-Converting Enzyme 2 as a Prognostic Biomarker for Disease Progression in Patients Infected with SARS-CoV-2.

Author

Díaz-Troyano N, Gabriel-Medina P, Weber S, Klammer M, Barquín-DelPino R, Castillo-Ribelles L, Esteban A, Hernández-González M, Ferrer-Costa R, Pumarola T, and Rodríguez-Frías F

Abstract

Predicting disease severity in patients infected with SARS-CoV-2 is difficult. Soluble angiotensin-converting enzyme 2 (sACE2) arises from the shedding of membrane ACE2 (mACE2), which is a receptor for SARS-CoV-2 spike protein. We evaluated the predictive value of sACE2 compared with known biomarkers of inflammation and tissue damage (CRP, GDF-15, IL-6, and sFlt-1) in 850 patients with and without SARS-CoV-2 with different clinical outcomes. For univariate analyses, median differences between biomarker levels were calculated for the following patient groups (classified by clinical outcome): RT-PCR-confirmed SARS-CoV-2 positive (Groups 1−4); RT-PCR-confirmed SARS-CoV-2 negative following previous SARS-CoV-2 infection (Groups 5 and 6); and ‘SARS-CoV-2 unexposed’ patients (Group 7). Median levels of CRP, GDF-15, IL-6, and sFlt-1 were significantly higher in hospitalized patients with SARS-CoV-2 compared with discharged patients (all p < 0.001), whereas levels of sACE2 were significantly lower (p < 0.001). ROC curve analysis of sACE2 provided cut-offs for predicting hospital admission (≤0.05 ng/mL (positive predictive value: 89.1%) and ≥0.42 ng/mL (negative predictive value: 84.0%)). These findings support further investigation of sACE2, as a single biomarker or as part of a panel, to predict hospitalization risk and disease severity in patients with SARS-CoV-2 infection.

Published
2022
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6. Utility of Transient Elastography for the Screening of Liver Disease in Patients with Alpha1-Antitrypsin Deficiency.

Author

Pons M, Núñez A, Esquinas C, Torres-Durán M, Rodríguez-Hermosa JL, Calle M, Tubio-Pérez R, Belmonte I, Rodríguez-Frías F, Rodríguez E, Genescà J, Miravitlles M, and Barrecheguren M

Abstract

Screening of liver disease in alpha-1 antitrypsin deficiency (AATD) is usually carried out with liver enzymes, with low sensitivity. We conducted a multicenter cross-sectional study aiming to describe the utility of transient elastography for the identification of liver disease in patients with AATD. A total of 148 AATD patients were included. Among these, 54.7% were Pi*ZZ and 45.3% were heterozygous for the Z allele. Between 4.9% and 16.5% of patients had abnormal liver enzymes, without differences among genotypes. Liver stiffness measurement (LSM) was significantly higher in Pi*ZZ individuals than in heterozygous Z (5.6 vs. 4.6 kPa; p = 0.001). In total, in 8 (5%) individuals LSM was >7.5 kPa, considered significant liver fibrosis, and ≥10 kPa in 3 (1.9%) all being Pi*ZZ. Elevated liver enzymes were more frequently observed in patients with LSM > 7.5 kPa, but in 5 out of 8 of these patients all liver enzymes were within normal range. In patients with AATD, the presence of abnormal liver enzymes is frequent; however, most of these patients do not present significant liver fibrosis. Transient elastography can help to identify patients with liver fibrosis even with normal liver enzymes and should be performed in all Z-allele carriers to screen for liver disease.

Published
2021
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