Identification of thyroid tumor cell vulnerabilities through a siRNA-based functional screening

The incidence of thyroid carcinoma is rapidly increasing. Although generally associated with good prognosis, a fraction of thyroid tumors are not cured by standard therapy and progress to aggressive forms for which no effective treatments are currently available. In order to identify novel therapeutic targets for thyroid carcinoma, we focused on the discovery of genes essential for sustaining the oncogenic phenotype of thyroid tumor cells, but not required to the same degree for the viability of normal cells (non-oncogene addiction paradigm). We screened a siRNA oligonucleotide library targeting the human druggable genome in thyroid cancer BCPAP cell line in comparison with immortalized normal human thyrocytes (Nthy-ori 3-1). We identified a panel of hit genes whose silencing interferes with the growth of tumor cells, while sparing that of normal ones. Further analysis of three selected hit genes, namely Cyclin D1, MASTL and COPZ1, showed that they represent common vulnerabilities for thyroid tumor cells, as their inhibition reduced the viability of several thyroid tumor cell lines, regardless the histotype or oncogenic lesion. This work identified non-oncogenes essential for sustaining the phenotype of thyroid tumor cells, but not of normal cells, thus suggesting that they might represent promising targets for new therapeutic strategies

Severity and prognosis in stroke: scoping review

Introdução: O Acidente Vascular Encefálico (AVE) tem vindo a apresentar uma taxa de incidência estável e um considerável declínio na taxa de mortalidade, o que corresponde a um aumento na prevalência de sobreviventes. O conhecimento das alterações funcionais que podem surgir após o AVE, da sua severidade e das estratégias disponíveis para avaliar a disfunção, facilita a construção de um plano de reabilitação, com objetivos para os profissionais de saúde, para os indivíduos e para a família dentro do potencial de recuperação. A severidade surge, como um conceito abrangente associado à presença de défices neurológicos, motores ou funcionais e às alterações das atividades da vida diária. Alterações neurológicas, motoras ou funcionais mais severas fazem prever uma recuperação mais difícil e mais prolongada. A determinação do prognóstico em indivíduos com AVE engloba não só o risco de morte a curto prazo como também a probabilidade de recuperar a função a longo prazo. Objetivo: Avaliar o panorama acerca da informação existente sobre o nível de severidade e prognóstico em AVE’s. Métodos: A Revisão Scoping baseou-se na metodologia de Arksey & O’Malley (2005), sendo constituída por seis passos: 1) Identificação da questão; 2) Identificação da literatura relevante; 3) Seleção da literatura; 4) Mapeamento dos dados; 5) Recolha, sumário e transcrição dos resultados; 6) Consultoria (opcional). Resultados: Foram analisados 47 estudos observacionais. 95% dos autores referem-se à severidade como sendo a quantidade de défices neurológicos apresentados pelos indivíduos após o AVE e avaliam-na através de instrumentos de medida específicos para a avaliação de défices neurológicos (76% dos autores utilizaram a NIHSS na sua metodologia). O prognóstico no AVE surge associado à funcionalidade alcançada (89%); probabilidade/índice de mortalidade (54%); e encaminhamento após a alta (15%). O prognóstico pode ser influenciado por fatores sociodemográficos, fatores clínicos e por algumas comorbilidades, entre outros. Conclusão: Os estudos de severidade e prognóstico em AVE’s poderão não refletir a condição real do indivíduo e induzir em erro a aplicação destes conceitos na prática clínica, influenciando o prognóstico esperado.ABSTRACT: Background – Stroke has shown a stable incidence rate and an important decrease in mortality rate, which corresponds to an increase in the survival prevalence. Knowledge of functional changes, stroke severity, and strategies to evaluate dysfunction after stroke, ease the conception of a rehabilitation plan, with objectives for health professionals, stroke patients, and their families. Stroke severity is related to neurologic, motor, function and daily activities changes. More severe neurological, motor or functional abnormalities predict a more difficult and longer recovery. Prognosis determination in stroke patients encompasses not only the risk of death in the short term but also the probability of recovering function in the long term. Aim(s) – To evaluate the existing information about stroke severity and prognosis. Methods – The scoping review was based on six steps Arksey & O’Malley (2005) methodology: 1) identifying the research question; 2) systematic search; 3) selection of publications; 4) charting the data; 5) collating, summarizing and reporting the results; 6) consultation (optional). Results – A total of 47 observational studies were analyzed. For 95% of authors, severity is the number of neurological deficits presented by individuals after stroke and was assessed through specific measurement instruments for neurological deficits (76% of authors used NIHSS in their methodology). Stroke prognosis is related to the functionality affected (89%); the probability of dead/mortality rate (54%); and referral after discharge (15%). Prognosis may be influenced by socio-demographic factors, clinical factors and by some comorbidity, among others. Conclusion – Severity and prognostic studies in stroke may not reflect the individual’s actual condition and mislead the use of these concepts, in reality, influencing the expected prognosis.info:eu-repo/semantics/publishedVersio

Haplotype Affinities Resolve a Major Component of Goat (Capra hircus) MtDNA D-Loop Diversity and Reveal Specific Features of the Sardinian Stock

Goat mtDNA haplogroup A is a poorly resolved lineage absorbing most of the overall diversity and is found in locations as distant as Eastern Asia and Southern Africa. Its phylogenetic dissection would cast light on an important portion of the spread of goat breeding. The aims of this work were 1) to provide an operational definition of meaningful mtDNA units within haplogroup A, 2) to investigate the mechanisms underlying the maintenance of diversity by considering the modes of selection operated by breeders and 3) to identify the peculiarities of Sardinian mtDNA types. We sequenced the mtDNA D-loop in a large sample of animals (1,591) which represents a non-trivial quota of the entire goat population of Sardinia. We found that Sardinia mirrors a large quota of mtDNA diversity of Western Eurasia in the number of variable sites, their mutational pattern and allele frequency. By using Bayesian analysis, a distance-based tree and a network analysis, we recognized demographically coherent groups of sequences identified by particular subsets of the variable positions. The results showed that this assignment system could be reproduced in other studies, capturing the greatest part of haplotype diversity

Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

Background: Many patients with COVID-19 have been treated with plasma containing anti-SARS-CoV-2 antibodies. We aimed to evaluate the safety and efficacy of convalescent plasma therapy in patients admitted to hospital with COVID-19. Methods: This randomised, controlled, open-label, platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]) is assessing several possible treatments in patients hospitalised with COVID-19 in the UK. The trial is underway at 177 NHS hospitals from across the UK. Eligible and consenting patients were randomly assigned (1:1) to receive either usual care alone (usual care group) or usual care plus high-titre convalescent plasma (convalescent plasma group). The primary outcome was 28-day mortality, analysed on an intention-to-treat basis. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936. Findings: Between May 28, 2020, and Jan 15, 2021, 11558 (71%) of 16287 patients enrolled in RECOVERY were eligible to receive convalescent plasma and were assigned to either the convalescent plasma group or the usual care group. There was no significant difference in 28-day mortality between the two groups: 1399 (24%) of 5795 patients in the convalescent plasma group and 1408 (24%) of 5763 patients in the usual care group died within 28 days (rate ratio 1·00, 95% CI 0·93–1·07; p=0·95). The 28-day mortality rate ratio was similar in all prespecified subgroups of patients, including in those patients without detectable SARS-CoV-2 antibodies at randomisation. Allocation to convalescent plasma had no significant effect on the proportion of patients discharged from hospital within 28 days (3832 [66%] patients in the convalescent plasma group vs 3822 [66%] patients in the usual care group; rate ratio 0·99, 95% CI 0·94–1·03; p=0·57). Among those not on invasive mechanical ventilation at randomisation, there was no significant difference in the proportion of patients meeting the composite endpoint of progression to invasive mechanical ventilation or death (1568 [29%] of 5493 patients in the convalescent plasma group vs 1568 [29%] of 5448 patients in the usual care group; rate ratio 0·99, 95% CI 0·93–1·05; p=0·79). Interpretation: In patients hospitalised with COVID-19, high-titre convalescent plasma did not improve survival or other prespecified clinical outcomes. Funding: UK Research and Innovation (Medical Research Council) and National Institute of Health Research

Tocilizumab in patients admitted to hospital with COVID-19 (RECOVERY): a randomised, controlled, open-label, platform trial

Background: In this study, we aimed to evaluate the effects of tocilizumab in adult patients admitted to hospital with COVID-19 with both hypoxia and systemic inflammation. Methods: This randomised, controlled, open-label, platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]), is assessing several possible treatments in patients hospitalised with COVID-19 in the UK. Those trial participants with hypoxia (oxygen saturation <92% on air or requiring oxygen therapy) and evidence of systemic inflammation (C-reactive protein ≥75 mg/L) were eligible for random assignment in a 1:1 ratio to usual standard of care alone versus usual standard of care plus tocilizumab at a dose of 400 mg–800 mg (depending on weight) given intravenously. A second dose could be given 12–24 h later if the patient's condition had not improved. The primary outcome was 28-day mortality, assessed in the intention-to-treat population. The trial is registered with ISRCTN (50189673) and ClinicalTrials.gov (NCT04381936). Findings: Between April 23, 2020, and Jan 24, 2021, 4116 adults of 21 550 patients enrolled into the RECOVERY trial were included in the assessment of tocilizumab, including 3385 (82%) patients receiving systemic corticosteroids. Overall, 621 (31%) of the 2022 patients allocated tocilizumab and 729 (35%) of the 2094 patients allocated to usual care died within 28 days (rate ratio 0·85; 95% CI 0·76–0·94; p=0·0028). Consistent results were seen in all prespecified subgroups of patients, including those receiving systemic corticosteroids. Patients allocated to tocilizumab were more likely to be discharged from hospital within 28 days (57% vs 50%; rate ratio 1·22; 1·12–1·33; p<0·0001). Among those not receiving invasive mechanical ventilation at baseline, patients allocated tocilizumab were less likely to reach the composite endpoint of invasive mechanical ventilation or death (35% vs 42%; risk ratio 0·84; 95% CI 0·77–0·92; p<0·0001). Interpretation: In hospitalised COVID-19 patients with hypoxia and systemic inflammation, tocilizumab improved survival and other clinical outcomes. These benefits were seen regardless of the amount of respiratory support and were additional to the benefits of systemic corticosteroids. Funding: UK Research and Innovation (Medical Research Council) and National Institute of Health Research

Abeta peptides accelerate the senescence of endothelial cells in vitro and in vivo, impairing angiogenesis

Cerebral amyloid angiopathy (CAA) caused by amyloid beta (Abeta) deposition around brain microvessels results in vascular degenerative changes. Antiangiogenic Abeta properties are known to contribute to the compromised cerebrovascular architecture. Here we hypothesize that Abeta peptides impair angiogenesis by causing endothelial cells to enter senescence at an early stage of vascular development. Wild-type (WT) Abeta and its mutated variant E22Q peptide, endowed with marked vascular tropism, were used in this study. In vivo, in zebrafish embryos, the WT or E22Q peptides reduced embryo survival with an IC(50) of 6.1 and 4.7 microM, respectively. The 2.5 microM concentration, showing minimal toxicity, was chosen. Alkaline phosphatase staining revealed disorganized vessel patterning, narrowing, and reduced branching of vessels. Beta-galactosidase staining and the cyclin-dependent kinase inhibitor p21 expression, indicative of senescence, were increased. In vitro, WT and E22Q reduced endothelial cell survival with an IC(50) of 12.3 and 8.8 microM, respectively. The 5 microM concentration, devoid of acute effects on the endothelium, was applied chronically to long-term cultured human umbilical vein endothelial cells (HUVECs). We observed reduced cumulative population doubling, which coincided with beta-galactosidase accumulation, down-regulation of telomerase reverse-transcriptase mRNA expression, decreased telomerase activity, and p21 activation. Senescent HUVECs showed marked angiogenesis impairment, as Abeta treatment reduced tube sprouting. The endothelial injuries caused by the E22Q peptide were much more aggressive than those induced by the WT peptide. Premature Abeta-induced senescence of the endothelium, producing progressive alterations of microvessel morphology and functions, may represent one of the underlying mechanisms for sporadic or heritable CAA

A\u3b2 peptides accelerate the senescence of endothelial cells in vitro and in vivo, impairing angiogenesis

Cerebral amyloid angiopathy (CAA) caused by amyloid beta (A beta) deposition around brain microvessels results in vascular degenerative changes. Antiangiogenic A beta properties are known to contribute to the compromised cerebrovascular architecture. Here we hypothesize that A beta peptides impair angiogenesis by causing endothelial cells to enter senescence at an early stage of vascular development. Wild-type (WT) A beta and its mutated variant E22Q peptide, endowed with marked vascular tropism, were used in this study. In vivo, in zebrafish embryos, the WT or E22Q peptides reduced embryo survival with an IC50 of 6.1 and 4.7 mu M, respectively. The 2.5 mu M concentration, showing minimal toxicity, was chosen. Alkaline phosphatase staining revealed disorganized vessel patterning, narrowing, and reduced branching of vessels. beta-Galactosidase staining and the cyclin-dependent kinase inhibitor p21 expression, indicative of senescence, were increased. In vitro, WT and E22Q reduced endothelial cell survival with an IC50 of 12.3 and 8.8 mu M, respectively. The 5 mu M concentration, devoid of acute effects on the endothelium, was applied chronically to long-term cultured human umbilical vein endothelial cells (HUVECs). We observed reduced cumulative population doubling, which coincided with beta-galactosidase accumulation, down-regulation of telomerase reverse-transcriptase mRNA expression, decreased telomerase activity, and p21 activation. Senescent HUVECs showed marked angiogenesis impairment, as A beta treatment reduced tube sprouting. The endothelial injuries caused by the E22Q peptide were much more aggressive than those induced by the WT peptide. Premature A beta-induced senescence of the endothelium, producing progressive alterations of microvessel morphology and functions, may represent one of the underlying mechanisms for sporadic or heritable CAA