Contribution of Dynamic and Genetic Tests for Short Stature Diagnosing: A Case Report

Genetics; Macimorelin; Short statureGenètica; Macimorelina; Baixa estaturaGenética; Macimorelina; Baja estaturaGenetic tests have led to the discovery of many novel genetic variants related to growth failure, but the clinical significance of some results is not always easy to establish. The aim of this report is to describe both clinical phenotype and genetic characteristics in an adult patient with short stature associated with a homozygous variant in disintegrin and metalloproteinase with thrombospondin motifs type 17 gene (ADAMTS17) combined with a homozygous variant in the GH secretagogue receptor (GHS-R). The index case had severe short stature (SS) (−3.0 SD), small hands and feet, associated with eye disturbances. Genetic tests revealed homozygous compounds for ADAMTS17 responsible for Weill–Marchesani-like syndrome but a homozygous variant in GHS-R was also detected. Dynamic stimulation with an insulin tolerance test showed a normal elevation of GH, while the GH response to macimorelin stimulus was totally flattened. We show the implication of the GHS-R variant and review the molecular mechanisms of both entities. These results allowed us to better interpret the phenotypic spectrum, associated co-morbidities, its implications in dynamic tests, genetic counselling and treatment options not only to the index case but also for her relatives

Functional Insight into and Refinement of the Genomic Boundaries of the JARID2-Neurodevelopmental Disorder Episignature

DNA methylation; Episignature; Intellectual disabilityMetilació de l'ADN; Episignatura; Discapacitat intel·lectualMetilación del ADN; Epifirma; Discapacidad intelectualJARID2 (Jumonji, AT-rich interactive domain 2) haploinsufficiency is associated with a clinically distinct neurodevelopmental syndrome. It is characterized by intellectual disability, developmental delay, autistic features, behavior abnormalities, cognitive impairment, hypotonia, and dysmorphic features. JARID2 acts as a transcriptional repressor protein that is involved in the regulation of histone methyltransferase complexes. JARID2 plays a role in the epigenetic machinery, and the associated syndrome has an identified DNA methylation episignature derived from sequence variants and intragenic deletions involving JARID2. For this study, our aim was to determine whether patients with larger deletions spanning beyond JARID2 present a similar DNA methylation episignature and to define the critical region involved in aberrant DNA methylation in 6p22–p24 microdeletions. We examined the DNA methylation profiles of peripheral blood from 56 control subjects, 13 patients with (likely) pathogenic JARID2 variants or patients carrying copy number variants, and three patients with JARID2 VUS variants. The analysis showed a distinct and strong differentiation between patients with (likely) pathogenic variants, both sequence and copy number, and controls. Using the identified episignature, we developed a binary model to classify patients with the JARID2-neurodevelopmental syndrome. DNA methylation analysis indicated that JARID2 is the driver gene for aberrant DNA methylation observed in 6p22–p24 microdeletions. In addition, we performed analysis of functional correlation of the JARID2 genome-wide methylation profile with the DNA methylation profiles of 56 additional neurodevelopmental disorders. To conclude, we refined the critical region for the presence of the JARID2 episignature in 6p22–p24 microdeletions and provide insight into the functional changes in the epigenome observed when regulation by JARID2 is lost.Funding for this study is provided in part by the Government of Canada through Genome Canada and the Ontario Genomics Institute (OGI-188)

The evolution of the ventilatory ratio is a prognostic factor in mechanically ventilated COVID-19 ARDS patients

Background: Mortality due to COVID-19 is high, especially in patients requiring mechanical ventilation. The purpose of the study is to investigate associations between mortality and variables measured during the first three days of mechanical ventilation in patients with COVID-19 intubated at ICU admission. Methods: Multicenter, observational, cohort study includes consecutive patients with COVID-19 admitted to 44 Spanish ICUs between February 25 and July 31, 2020, who required intubation at ICU admission and mechanical ventilation for more than three days. We collected demographic and clinical data prior to admission; information about clinical evolution at days 1 and 3 of mechanical ventilation; and outcomes. Results: Of the 2,095 patients with COVID-19 admitted to the ICU, 1,118 (53.3%) were intubated at day 1 and remained under mechanical ventilation at day three. From days 1 to 3, PaO2/FiO2 increased from 115.6 [80.0-171.2] to 180.0 [135.4-227.9] mmHg and the ventilatory ratio from 1.73 [1.33-2.25] to 1.96 [1.61-2.40]. In-hospital mortality was 38.7%. A higher increase between ICU admission and day 3 in the ventilatory ratio (OR 1.04 [CI 1.01-1.07], p = 0.030) and creatinine levels (OR 1.05 [CI 1.01-1.09], p = 0.005) and a lower increase in platelet counts (OR 0.96 [CI 0.93-1.00], p = 0.037) were independently associated with a higher risk of death. No association between mortality and the PaO2/FiO2 variation was observed (OR 0.99 [CI 0.95 to 1.02], p = 0.47). Conclusions: Higher ventilatory ratio and its increase at day 3 is associated with mortality in patients with COVID-19 receiving mechanical ventilation at ICU admission. No association was found in the PaO2/FiO2 variation

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Anàlisi molecular dels gens "BRCA1" i "BRCA2" en el càncer de mama hereditari. Caracterització de mutacions recurrents i estudi de variants d'efecte biològic desconegut

El càncer de mama és la neoplàsia més freqüent en la dona a Catalunya, amb una prevalença de 45.000 casos l'any 2005. Entre un 5% i un 10% de tots els casos presenta un component hereditari degut a mutacions germinals en gens de susceptibilitat, fonamentalment BRCA1 i BRCA2, els quals s'associen a la síndrome de càncer de mama i ovari hereditari (CMOH), caracteritzada per la presència de casos de càncer de mama i ovari en una mateixa família o de múltiples casos de càncer de mama precoç. L'objectiu principal d'aquest treball és l'estudi molecular dels gens BRCA1 i BRCA2 en pacients amb sospita de síndrome de CMOH. La identificació de mutacions en aquests pacients facilita l'assessorament genètic i l'aplicació de mesures de prevenció, detecció precoç i terapèutiques adequades.Els mètodes utilitzats per a la detecció de mutacions en BRCA1 i BRCA2 són l'anàlisi de conformacions de cadena senzilla (SSCP), l'anàlisi de la proteïna truncada (PTT) i la seqüenciació dels fragments amb mobilitat electroforètica anormal. Els nostres resultats mostren que les famílies espanyoles amb CMOH presenten una gran diversitat al·lèlica en els gens BRCA1 i BRCA2, amb les mutacions distribuïdes al llarg de tota la seqüència. La freqüència global de mutacions identificades (propera al 30%) és similar a la trobada en altres poblacions i, en general, el percentatge de mutacions augmenta amb el nombre de casos de càncer en la família. A més, s'evidencia la relació entre càncer d'ovari i BRCA1 i càncer de mama en homes i BRCA2. Addicionalment, s'ha analitzat l'haplotip associat a tres mutacions que apareixen de forma recurrent en diverses famílies, mitjançant l'estudi de marcadors microsatèl·lits polimòrfics. La identificació d'haplotips específics associats a la mutació 330A>G en BRCA1, present en famílies d'origen gallec, i a les mutacions 9254del5 i 6857delAA en BRCA2, presents en famílies de l'àrea mediterrània, suggereix l'existència de subpoblacions espanyoles amb mutacions característiques.Finalment, s'ha avaluat la possible patogenicitat de les variants d'efecte biològic desconegut identificades durant el procés de detecció de mutacions. Aquestes variants no es poden classificar, únicament amb la informació que ens proporciona l'estudi del DNA, com a mutacions associades a la malaltia o com a polimorfismes innocus, de manera que dificulten l'assessorament genètic dels pacients i requereixen anàlisis addicionals. Per exemple, l'anàlisi de l'RNA missatger de portadors permet establir possibles efectes en el procés de maduració de l'RNA (splicing). Malgrat que existeixen diversos mètodes teòrics per predir potencials alteracions en l'splicing, els resultats observats amb l'estudi de l'RNA no coincideixen totalment amb els obtinguts amb aquests mètodes, pel que s'han de considerar amb prudència i es recomana l'anàlisi de l'RNA sempre que sigui possible.Breast cancer is the most common malignancy among woman in Catalonia, with a prevalence of 45.000 cases in 2005. It is currently estimated that 5-10% of all breast cancers are hereditary and attributable to mutations in several highly penetrant susceptibility genes of which only two have been identified: BRCA1 and BRCA2. BRCA1 and BRCA2 mutations are associated with the hereditary breast and ovarian cancer (HBOC)syndrome, characterized for the presence of both breast and ovarian cases in the same family or for several cases of early onset breast cancer. The principal aim of this thesis is the molecular study of BRCA1 and BRCA2 genes in patients with HBOC syndrome. The identification of mutations in these patients facilitates the genetic assessment and the application of preventive measures, early detection and appropriate therapeutics. Analyses of the entire coding and flanking sequences were carried out by a combination of SSCP (single strand conformation polymorphism) and PTT (protein truncation test), followed by direct sequencing of abnormal bands. Our results show that HBOC Spanish families present a high allelic diversity in BRCA genes, with mutations distributed all over the sequence. The global frequency of mutations detected (of about 30%) is similar to that founded in other populations and, in general, the rate of mutations increases with the number of cancer cases in the family. Furthermore, ovarian cancer associates to BRCA1 and male breast cancer to BRCA2.Additionally, we analysed the haplotype associated to three mutations which appear recurrently in several families, employing microsatellite markers. Identification of a specific haplotype associated to the 330A>G mutation in BRCA1, present in Galician families, and to the 9254del5 and 6857delAA mutations in BRCA2, present in Mediterranean families, suggests the existence of Spanish subpopulations with characteristic mutations.Finally, we evaluated the pathological effect of variants of unknown pathological significance by RNA analysis. This information is essential for providing efficient counselling for breast/ovarian cancer families

Molecular analysis of BRCA1 and BRCA2 genes in the hereditary breast cancer. Characterization of recurrent mutations and study of unclassified variants

[cat] El càncer de mama és la neoplàsia més freqüent en la dona a Catalunya, amb una prevalença de 45.000 casos l'any 2005. Entre un 5% i un 10% de tots els casos presenta un component hereditari degut a mutacions germinals en gens de susceptibilitat, fonamentalment BRCA1 i BRCA2, els quals s'associen a la síndrome de càncer de mama i ovari hereditari (CMOH), caracteritzada per la presència de casos de càncer de mama i ovari en una mateixa família o de múltiples casos de càncer de mama precoç. L'objectiu principal d'aquest treball és l'estudi molecular dels gens BRCA1 i BRCA2 en pacients amb sospita de síndrome de CMOH. La identificació de mutacions en aquests pacients facilita l'assessorament genètic i l'aplicació de mesures de prevenció, detecció precoç i terapèutiques adequades. Els mètodes utilitzats per a la detecció de mutacions en BRCA1 i BRCA2 són l'anàlisi de conformacions de cadena senzilla (SSCP), l'anàlisi de la proteïna truncada (PTT) i la seqüenciació dels fragments amb mobilitat electroforètica anormal. Els nostres resultats mostren que les famílies espanyoles amb CMOH presenten una gran diversitat al·lèlica en els gens BRCA1 i BRCA2, amb les mutacions distribuïdes al llarg de tota la seqüència. La freqüència global de mutacions identificades (propera al 30%) és similar a la trobada en altres poblacions i, en general, el percentatge de mutacions augmenta amb el nombre de casos de càncer en la família. A més, s'evidencia la relació entre càncer d'ovari i BRCA1 i càncer de mama en homes i BRCA2. Addicionalment, s'ha analitzat l'haplotip associat a tres mutacions que apareixen de forma recurrent en diverses famílies, mitjançant l'estudi de marcadors microsatèl·lits polimòrfics. La identificació d'haplotips específics associats a la mutació 330A>G en BRCA1, present en famílies d'origen gallec, i a les mutacions 9254del5 i 6857delAA en BRCA2, presents en famílies de l'àrea mediterrània, suggereix l'existència de subpoblacions espanyoles amb mutacions característiques. Finalment, s'ha avaluat la possible patogenicitat de les variants d'efecte biològic desconegut identificades durant el procés de detecció de mutacions. Aquestes variants no es poden classificar, únicament amb la informació que ens proporciona l'estudi del DNA, com a mutacions associades a la malaltia o com a polimorfismes innocus, de manera que dificulten l'assessorament genètic dels pacients i requereixen anàlisis addicionals. Per exemple, l'anàlisi de l'RNA missatger de portadors permet establir possibles efectes en el procés de maduració de l'RNA (splicing). Malgrat que existeixen diversos mètodes teòrics per predir potencials alteracions en l'splicing, els resultats observats amb l'estudi de l'RNA no coincideixen totalment amb els obtinguts amb aquests mètodes, pel que s'han de considerar amb prudència i es recomana l'anàlisi de l'RNA sempre que sigui possible.[eng] Breast cancer is the most common malignancy among woman in Catalonia, with a prevalence of 45.000 cases in 2005. It is currently estimated that 5-10% of all breast cancers are hereditary and attributable to mutations in several highly penetrant susceptibility genes of which only two have been identified: BRCA1 and BRCA2. BRCA1 and BRCA2 mutations are associated with the hereditary breast and ovarian cancer (HBOC) syndrome, characterized for the presence of both breast and ovarian cases in the same family or for several cases of early onset breast cancer. The principal aim of this thesis is the molecular study of BRCA1 and BRCA2 genes in patients with HBOC syndrome. The identification of mutations in these patients facilitates the genetic assessment and the application of preventive measures, early detection and appropriate therapeutics. Analyses of the entire coding and flanking sequences were carried out by a combination of SSCP (single strand conformation polymorphism) and PTT (protein truncation test), followed by direct sequencing of abnormal bands. Our results show that HBOC Spanish families present a high allelic diversity in BRCA genes, with mutations distributed all over the sequence. The global frequency of mutations detected (of about 30%) is similar to that founded in other populations and, in general, the rate of mutations increases with the number of cancer cases in the family. Furthermore, ovarian cancer associates to BRCA1 and male breast cancer to BRCA2. Additionally, we analysed the haplotype associated to three mutations which appear recurrently in several families, employing microsatellite markers. Identification of a specific haplotype associated to the 330A>G mutation in BRCA1, present in Galician families, and to the 9254del5 and 6857delAA mutations in BRCA2, present in Mediterranean families, suggests the existence of Spanish subpopulations with characteristic mutations. Finally, we evaluated the pathological effect of variants of unknown pathological significance by RNA analysis. This information is essential for providing efficient counselling for breast/ovarian cancer families