The transcriptome of the Oncogenic HOXA9 homeoprotein in human glioblastoma : functional and clinical relevance

Dissertação de mestrado em Genética MolecularGliomas are a heterogeneous group of neoplasias that account for the majority of primary tumors of the central nervous system, of which glioblastoma (GBM) is the most common and malignant subtype. These are dramatic diseases for which no curative therapies are yet available. The clinical responses of GBM patients are particularly poor and vary greatly among individuals, especially due to the heterogeneity of their molecular alterations. Regardless, these patients are equally treated with a standardized therapeutic approach, mainly due to the lack of wellestablished molecular prognostic markers. Recently, the reactivation of HOXA9 expression in GBM has been implicated as a poor prognostic marker. As HOXA9 is a transcription factor, we hypothesized that a set of HOXA9-transcriptionally regulated genes may be its true biological effectors. In this sense, we aimed to characterize the HOXA9 transcriptome in GBM to identify novel prognostic biomarkers and putative therapeutic targets. By analyzing expression microarrays in HOXA9-negative and positive U87MG and hTERT/E6/E7 cells, we found a vast number of genes regulated by HOXA9 that are involved in important hallmarks of cancer as proliferation, invasion, and therapy resistance. Interestingly, we found high expression of the long non-coding RNA HOTAIR (HOX transcript antisense intergenic RNA) in HOXA9-positive GBM cell lines, consistent with a significant co-expression between HOTAIR and HOXA9 in highgrade gliomas, particularly GBMs. Mechanistically, using chromatin immunoprecipitation and quantitative PCR analysis in GBM cell lines, we found that HOXA9 directly interacts with the promoter region of HOTAIR and induces its transcription. Importantly, GBM patients with high expression of HOTAIR had a relatively shorter overall survival, independently of other putative prognostic factors. Using in silico analysis we found other putative direct targets of HOXA9, and ChIP analysis proved the direct regulation of WNT6 (Wingless-Type MMTV Integration Site Family Member 6) by HOXA9. Our study provides the first characterization of HOXA9 target genes in the context of GBM, and identifies new clinically-relevant prognostic biomarkers, which may be new therapeutic targets to treat this aggressive malignancy.Os gliomas são um grupo heterogéneo de neoplasias que constituem a maioria dos tumores primários do sistema nervoso central, dos quais o glioblastoma (GBM) é o simultaneamente o mais comum e o mais maligno. Actualmente não existem ainda terapias eficazes para estas neoplasias altamente agressivas. A resposta clínica dos pacientes de GBM é extremamente insatisfatória e variável entre indivíduos, sobretudo devido à heterogeneidade das alterações moleculares presente nestes tumores. Devido à falta de factores de prognóstico bem estabelecidos, todos os pacientes são tratados com a mesma abordagem terapêutica. Recentemente, a reactivação da expressão do HOXA9 em GBM foi implicada como um marcador de pior prognóstico. Como o HOXA9 é um factor de transcrição, colocamos a hipótese de que um grupo de genes regulados pelo HOXA9 poderão ser os seus efetores biológicos. Assim, tivemos como objectivo caracterizar o transcriptoma do HOXA9 em GBM, para identificar novos biomarcadores de prognóstico e potenciais alvos terapêuticos. Pela análise de microarrays de expressão em linhas celulares U87MG e hTERT/E6/E7, com e sem a expressão de HOXA9, verificámos que muitos dos genes-alvo do HOXA9 regulam características críticas no cancro, como a proliferação, a invasão e a resistência à terapia. Verificámos ainda uma expressão aumentada do RNA não-codificante HOTAIR (HOX transcript antisense intergenic RNA) em células de GBM HOXA9-positivas, um resultado consistente com a co-expressão entre HOTAIR e HOXA9 em gliomas primários de alto-grau. Usando imunoprecipitação da cromatina (ChIP) e PCR quantitativo em linhas celulares de GBM verificámos que o HOXA9 interage directamente com a região promotora do HOTAIR, induzindo a sua expressão. Encontrámos também uma relevante associação entre altos níveis de expressão do HOTAIR e uma pior sobrevida dos pacientes de GBM. Por análises in silico, encontrámos outros potenciais alvos directos do HOXA9, tendo validado o WNT6 (Wingless-Type MMTV Integration Site Family Member 6) por ChIP. Resumindo, o nosso estudo providencia a primeira caracterização dos alvos do HOXA9 em GBM, e identifica novos biomarcadores de prognóstico, que poderão auxiliar a racionalização das decisões terapêuticas, bem como vir a ser testados como novos alvos terapêuticos

Influence of HOTAIR rs920778 and rs12826786 genetic variants on prostate cancer risk and progression-free survival

Aim: Evaluate the impact of the single nucleotide polymorphisms rs920778 and rs12826786 in the long noncoding RNA HOTAIR in the susceptibility and prognosis of prostate cancer (PCa) patients. Patients & methods: HOTAIR single nucleotide polymorphisms were genotyped by restriction fragment length polymorphism in 151 PCa cases and 180 cancer-free controls. Odds ratio, 95% CIs and prognostic significance were calculated. Results: Our data showed no statistically significant associations between HOTAIR polymorphic variants in rs920778 and rs12826786 and PCa susceptibility. However, the CC genotype in rs12826786 was significantly associated with shorter biochemical recurrence-free survival in pT3-stage PCa patients. Conclusion: Our results indicate that HOTAIR rs12826786 CC genotype may be an independent prognostic biomarker in a particular subset of PCa tumors.Fundação para a Ciência e Tecnologia (IF/00601/2012 to BM Costa; SFRH/BD/52287/2013 to AI Oliveira; SFRH/BD/88220/2012 to AX Magalhaes), Fundação Calouste Gulbenkian (BM Costa), Liga Portuguesa Contra o Cancro (BM Costa) and Inter-University Doctoral Programme in Ageing and Chronic Disease (PhDOC; to AI Oliveira and AX Magalhaes). Project co-financed by Programa Operacional Regional do Norte (ON.2-O Novo Norte), Quadro de Referencia Estratégico Nacional (QREN), Fundo Europeu de Desenvolvimento Regional (FEDER) and a grant (74-CI-IPOP) from Research Center of Portuguese Oncology Institute of Porto (C Jeronimo).info:eu-repo/semantics/publishedVersio

A novel molecular link between HOXA9 and WNT6 in glioblastoma identifies a subgroup of patients with particular poor prognosis

Despite much effort to improve treatments, patients with malignant glioma still present a very poor prognosis that has not changed significantly in the last decades. In this context, it is crucial to better understand glioma pathogenesis to identify new molecular prognostic subgroups and therapeutic targets. WNT6 was recently identified as a new oncogenic molecule in glioblastoma (GBM), with prognostic value in patients, but the mechanisms underlying WNT6 aberrant expression in glioma are still unknown. WNT6 was overexpressed in a subset of gliomas independently of IDH mutations, 1p/19q codeletion status, and WNT6 gene copy number. Interestingly, WNT6 expression is associated with the DNA methylation levels of particular CpG regions at both the WNT6 promoter and the gene body in glioma patient samples. HOXA9, a transcription factor previously associated with poorer clinical outcome in GBM, was identified as a novel transcriptional regulator of WNT6, activating the WNT/β-catenin pathway in vitro and in vivo. In various cohorts of glioma patients, mRNA levels of WNT6 and HOXA9 were significantly correlated, extending our in vitro and in vivo findings into the clinical setting. Interestingly, this novel molecular link between WNT6 and HOXA9 was not limited to glioma, as they were co-expressed also in patients with other tumor types. Clinically, WNT6 was a prognostic biomarker of shorter survival in GBM, independently of HOXA9 expression. Concomitant high expression of both WNT6 and HOXA9 identified a subgroup of patients with particularly dismal survival. These findings describe novel WNT6 regulatory mechanisms in GBM, establishing particular DNA methylation patterns and HOXA9 as critical regulators of WNT6 expression in glioma. This HOXA9-WNT6 molecular link supports WNT signaling in GBM cells and is a powerful prognostic biomarker, highlighting the clinical relevance of this axis in patients. Novel therapies targeting WNT6-HOXA9 signaling may thus be useful for this deadly disease.info:eu-repo/semantics/publishedVersio

EPIDEMIOLOGICAL PROFILE OF CASES OF SEXUAL VIOLENCE IN ANÁPOLIS - GOIÁS - BRAZIL, IN THE YEARS 2017 TO 2020

Objective: to identify the epidemiological profile of victims who suffered sexual violence, in Anápolis - Goiás - Brazil, between the years 2017 to 2020.Methods: descriptive study of quantitative nature. Data from the notification form of sexual violence registered in the Department of Epidemiological Surveillance were used. Chi-square test was used with a significance level of 5% (p<0.05). Results: 850 cases were notified, with the peak in 2020, 82.1% were female, between the age group of two to 10 years (42.2%), brown (61.2%) and single (37.1%). The main conduct carried out after sexual violence was blood collection (21.6%); the main aggressors were the parents (20.9%), the main type of sexual violence was rape (68.7%) and the victims were mainly referred to the Guardianship Council (48.2%).Conclusion: The results point to the need to strengthen interventions to expand the social protection network for children and adolescents

Round multiprofissional com checklist: associação com a melhoria na segurança do paciente em terapia intensiva

Objetivo: Verificar a associação entre round multiprofissional com uso de checklist e práticas de segurança do paciente por profissionais de saúde de uma unidade de terapia intensiva.Método: Estudo de método misto, delineado pela abordagem sequencial explanatória, realizado em um hospital do sul do Brasil. Os dados quantitativos foram analisados por meio de regressão de Poisson e os dados qualitativos, pela análise de conteúdo. Fez-se a análise integrada por meio da combinação explicada/conectada.Resultados: No período pós-implementação dos rounds com uso sistemático de checklist houve melhora significativa da profilaxia de tromboembolia venosa, sedação leve, redução dos dias de uso de ventilação mecânica, cateter venoso central e de sonda vesical de demora.Conclusão: O round multiprofissional com uso sistemático de checklist, associado com a melhoria nas práticas de segurança do paciente, foi considerado como uma estratégia que assegura melhores cuidados em terapia intensiva e favorece a satisfação no trabalho. Palavras-chave: Lista de checagem. Visitas com preceptor. Unidades de terapia intensiva. Segurança do paciente. Equipe de assistência ao paciente

Permanent health education in the context of obesity: a scoping review

OBJETIVE: To map the international literature on Permanent Health Education initiatives to care for people with obesity. METHODS: In total, six databases were searched without any language or publication period restriction according to the Joana Briggs Institute manual for evidence synthesis and the Prisma extension for scoping reviews (Prisma-ScR). Articles were independently analyzed by four reviewers and data, by two authors, which were then analyzed and discussed with our research team. RESULTS: After screening 8,780 titles/abstracts and 26 full texts, 10studies met our eligibility criteria. We extracted data on methodologies, themes, definitions of obesity, outcomes, and gaps. Most initiatives came from North American countries without free or universal health systems and lasted a short period of time (70%), had multidisciplinary teams (70%), and addressed sub-themes on obesity approaches (90%). Results included changes in participants’ understanding, attitude, and procedures (80%) and gaps which pointed to the sustainability of these changes (80%). CONCLUSION: This review shows the scarce research in the area and a general design of poorly effective initiatives, with traditional teaching methodologies based on information transmission techniques, the understanding of obesity as a disease and a public health problem, punctual actions, disciplinary fragmentation alien to the daily work centrality, and failure to recognize problems and territory as knowledge triggers and to focus on health care networks, line of care, the integrality of care, and food and body cultures

HOXA9 promotes glioblastoma initiation, aggressiveness and resistance to therapy

Glioblastoma is the most common and malignant subtype of glioma, exhibiting remarkable resistance to treatment. Here we investigated the oncogenic potential of HOXA9 in gliomagenesis, the molecular and cellular mechanisms by which HOXA9 may render glioblastoma more aggressive, and how HOXA9 affects response to chemotherapy and prognosis. Expression microarrays were used to identify HOXA9 target genes. Stable glioblastoma cell lines with ectopic HOXA9 overexpression or shRNA-­mediated knockdown of HOXA9 were established to evaluate the roles of HOXA9 in cell viability, death, invasion, and response to temozolomide. Subcutaneous and orthotopic intracranial xenograft models of glioblastoma were established to evaluate the oncogenic potential of HOXA9 in vivo, and its role in response to temozolomide and overall survival. Transcriptomic analyses identified novel HOXA9-­target genes that have key roles in critical cancer processes, including cell proliferation, adhesion, DNA metabolism and repair, and stem cell maintenance. Functional assays with a variety of glioblastoma cells revealed that HOXA9 promotes cell viability, stemness, and invasion; conversely, HOXA9 displayed anti-­apoptotic functions. Additionally, ectopic expression of HOXA9 promoted the malignant transformation of human immortalized astrocytes in an intracranial orthotopic mouse model of glioblastoma, and caused tumor-­associated death. HOXA9 also mediated resistance to temozolomide treatment both in vitro and in vivo. Mechanistically, BCL2 was identified as a novel HOXA9 target that may be therapeutically targeted. Indeed, the pharmacological inhibition of BCL2 with ABT-­737 specifically reverted temozolomide resistance in HOXA9-­positive cells. These data establish HOXA9 as a critical driver of glioma initiation, aggressiveness and resistance to therapy

Perfil clínico-epidemiológico dos casos de violência sexual em Anápolis-Goiás, entre os anos de 2016 a 2020

A violência sexual pode ser caracterizada como toda ação na qual uma pessoa, numa relação de poder, por meio de força física, coerção, sedução ou intimidação psicológica, obriga a outra pessoa a praticar ou submeter-se à relação sexual ou a situações que possam ferir a integridade física e/ou moral da vítima. No Brasil, a violência sexual é considerada uma situação alarmante, apresentando alto número de casos anualmente e, por esse motivo, é um evento de notificação compulsória nos serviços de saúde, sejam públicos ou privados. Essa notificação é realizada por meio do preenchimento de uma ficha de notificação, cujos dados são inseridos no Sistema de Informação de Agravos de Notificação (SINAN), um sistema de vigilância epidemiológica. O preenchimento dessa ficha é considerado essencial à análise epidemiológica, operacional e na construção do perfil do caso. Por isso, objetiva-se com esse estudo descrever o perfil clínico epidemiológico de vítimas que sofreram violência sexual, notificadas no SINAN, no município de Anápolis-GO, entre os anos 2016 a 2020. Trata-se de um estudo epidemiológico, observacional, descritivo, transversal e retrospectivo que tomará como fonte de informação os casos de violência sexual contra mulheres, crianças e adolescentes, de ambos os sexos vinculados no Departamento de Vigilância Epidemiológica da Secretaria Municipal de Saúde de Anápolis, o trabalho será aprovado pelo Comitê de Ética em Pesquisa– UniEVANGÉLICA seguindo a Resolução 466/2012 do Conselho Nacional de Saúde (CNS). A partir disso, esperase contribuir com a difusão de conhecimentos sobre a violência sexual, principais fatores influenciadores e suas principais consequências, com o planejamento e a implementação de políticas públicas e programas estratégicos de prevenção e intervenção deste agravo