The Barretos Cancer Hospital Animal Facility: implementation and results of a dedicated platform for preclinical oncology models

The Barretos Cancer Hospital Animal Facility (BCHAF) is a unique facility in Brazil exclusively dedicated to working with animal models for cancer research. In this article, we briefly present our modern facility and the main experiments performed, focusing on mutant strains of mice (PTCH-knockout and ApcMin mice), xenograft models, and patient-derived xenografts (PDXs). Our results show the progress and challenges in establishing these models and the need for having an appropriate representation of our cancer population to better understand tumor biology and to identify cancer biomarkers, which could be putatively targeted, allowing for personalized therapy.This study was funded by the Public Ministry of Labor Campinas (Research, Prevention and Education of Occupational Cancer) and by Pio XII Foundation, Barretos Cancer Hospital internal funds, Grant Number: 13/2021

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

Eficácia do [10]-gingerol contra metástases de câncer de mama : estudos in vitro e in vivo em camundongos

Breast cancer is the leading cause of death in Brazil and worldwide, between the types of cancer, triple negative TNBC (Triple Negative Breast Cancer), which do not express hormone receptors, represents 20% of the cases and presents relapses within 3 years. However, the major cause of death in cancer is due the formation of metastasis. The TNBC has a greater propensity to form lung and brain metastasis. Furthermore, currently few treatments are available. Search for new target treatments, with fewer side effects is very important to treat this disease. Natural products are rich sources of molecules with antitumoral activity; among them are the gingerols, from ginger and resveratrol, from grapes. Nevertheless, not much is known about antitumor activity of [10]-gingerol (10G). Therefore, the aim of this work was to investigate the potential use of 10G as an antimetastatic molecule in in vitro and in vivo experiments. In this work, 10G had antiproliferative activity on several breast cancer cell lines (4T1BM2, 4T1BM2 shRNAβ3, 4T1Br4, MDA-MB-231, MDA-MB-231 BrM) and on a normal cell line, bEnd.3. The natural compounds affected tumor cell morphology and RSVT was able to inhibit cell migration and adhesion through vitronectin. 10G induced apoptosis via the extrinsic pathway through the increase of caspase-9, -3 and -7 expression and decrease of Bcl-2 expression, induced cell cycle arrest in G1 and subG0 phase. In in vivo models, 10G inhibited primary tumor growth and lung metastasis, as well as bone and brain metastasis. Therefore, this study was able to provide new data as 10G can be acting in tumor cells and its possible clinical application.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)O câncer de mama é o que mais afeta as mulheres no Brasil e no mundo, entre os tipos de câncer de mama, o triplo negativo TNBC (Triple Negative Breast Cancer), que não expressa nenhum receptor hormonal, corresponde a 20% dos casos e apresenta recidivas em três anos. Entretanto, a maior causa de morte no câncer é devido a formação de metástases. O TNBC possui uma maior propensão a metástases pulmonares e cerebrais. Além disso, atualmente poucos tratamentos estão disponíveis. Buscas para novos medicamentos alvo para o TNBC e com menos efeitos colaterais são de grande importância para o tratamento dessa doença. Produtos naturais são fontes ricas em moléculas com atividades antitumorais, dentre elas os gingeróis, presentes no gengibre e o resveratrol (RSVT), presente nas uvas. Entretanto, pouco se sabe da atividade antitumoral do [10]-gingerol (10G). Este trabalho teve como objetivo investigar o potencial uso do 10G como molécula antimetastática em experimentos in vitro e in vivo. Neste trabalho verificou-se a atividade antiproliferativa do 10G e do RSVT em diversas linhagens de câncer de mama (4T1BM2, 4T1BM2 shRNAβ3, 4T1Br4, MDA-MB- 231, MDA-MB-231 BrM) e um controle de célula normal, a bEnd.3. Os produtos naturais (PNs) usados afetam a morfologia celular nas células tumorais e o RSVT é capaz de inibir a adesão e migração via vitronectina. O 10G induziu apoptose da linhagem 4T1Br4 através da via extrínseca, com o aumento da expressão de casspase-9, -3 e -7; induziu a parada do ciclo celular nas fases G1 e sub G0. Em modelos de metástase in vivo o 10G inibiu o crescimento do tumor primário e de metástases pulmonares, além de inibir metástases ósseas e cerebrais. Portanto, este trabalho foi capaz de fornecer dados de como o 10G atua nas células tumorais e sua possível aplicação clínica

ADAM9 silencing inhibits breast tumor cell invasion in vitro

AbstractADAM9 (A Disintegrin And Metalloproteinase 9) is a member of the ADAM protein family which contains a disintegrin domain. This protein family plays key roles in many physiological processes, including fertilization, migration, and cell survival. The ADAM proteins have also been implicated in various diseases, including cancer. Specifically, ADAM9 has been suggested to be involved in metastasis. To address this question, we generated ADAM9 knockdown clones of MDA-MB-231 breast tumor cells using silencing RNAs that were tested for cell adhesion, proliferation, migration and invasion assays. In RNAi-mediated ADAM9 silenced MDA-MB-231 cells, the expression of ADAM9 was lower from the third to the sixth day after silencing and inhibited tumor cell invasion in matrigel by approximately 72% when compared to control cells, without affecting cell adhesion, proliferation or migration. In conclusion, the generation of MDA-MB-231 knockdown clones lacking ADAM9 expression inhibited tumor cell invasion in vitro, suggesting that ADAM9 is an important molecule in the processes of invasion and metastasis

Identification of brain metastasis genes and therapeutic evaluation of histone deacetylase inhibitors in a clinically relevant model of breast cancer brain metastasis

Breast cancer brain metastasis remains largely incurable. While several mouse models have been developed to investigate the genes and mechanisms regulating breast cancer brain metastasis, these models often lack clinical relevance since they require the use of immune-compromised mice and/or are poorly metastatic to brain from the mammary gland. We describe the development and characterisation of an aggressive brain metastatic variant of the 4T1 syngeneic model (4T1Br4) that spontaneously metastasises to multiple organs, but is selectively more metastatic to the brain from the mammary gland than parental 4T1 tumours. By immunohistochemistry, 4T1Br4 tumours and brain metastases display a triple negative phenotype, consistent with the high propensity of this breast cancer subtype to spread to brain. assays indicate that 4T1Br4 cells have an enhanced ability to adhere to or migrate across a brain-derived endothelial monolayer and greater invasive response to brain-derived soluble factors compared to 4T1 cells. These properties are likely to contribute to the brain-selectivity of 4T1Br4 tumours. Expression profiling and gene set enrichment analyses demonstrate the clinical relevance of the 4T1Br4 model at the transcriptomic level. Pathway analyses implicate tumour-intrinsic immune regulation and vascular interactions in successful brain colonisation, revealing potential therapeutic targets. Evaluation of two histone deacetylase inhibitors, SB939 and 1179.4b, shows partial efficacy against 4T1Br4 metastasis to brain and other sites and potent radio-sensitising properties The 4T1Br4 model provides a clinically relevant tool for mechanistic studies and to evaluate novel therapies against brain metastasis

Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries

Background Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks. Methods The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned. Results A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P < 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31). Conclusion Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)

Measurement of Ξc+\Xi_{c}^{+} production in ppPb collisions at sNN=8.16\sqrt{s_{NN}}=8.16 TeV at LHCb

International audienceA study of prompt $\Xi_{c}^{+}$ production in proton-lead collisions is performed with the LHCb experiment at a centre-of-mass energy per nucleon pair of 8.16 TeV in 2016 in $p$Pb and Pb$p$ collisions with an estimated integrated luminosity of approximately 12.5 and 17.4 nb$^{-1}$, respectively. The $\Xi_{c}^{+}$ production cross-section, as well as the $\Xi_{c}^{+}$ to $\Lambda_{c}^{+}$ production cross-section ratio, are measured as a function of the transverse momentum and rapidity and compared to latest theory predictions. The forward-backward asymmetry is also measured as a function of the $\Xi_{c}^{+}$ transverse momentum

Measurement of the ratios of branching fractions R(D∗)\mathcal{R}(D^{*}) and R(D0)\mathcal{R}(D^{0})

International audienceThe ratios of branching fractions $\mathcal{R}(D^{*})\equiv\mathcal{B}(\bar{B}\to D^{*}\tau^{-}\bar{\nu}_{\tau})/\mathcal{B}(\bar{B}\to D^{*}\mu^{-}\bar{\nu}_{\mu})$ and $\mathcal{R}(D^{0})\equiv\mathcal{B}(B^{-}\to D^{0}\tau^{-}\bar{\nu}_{\tau})/\mathcal{B}(B^{-}\to D^{0}\mu^{-}\bar{\nu}_{\mu})$ are measured, assuming isospin symmetry, using a sample of proton-proton collision data corresponding to 3.0 fb${ }^{-1}$ of integrated luminosity recorded by the LHCb experiment during 2011 and 2012. The tau lepton is identified in the decay mode $\tau^{-}\to\mu^{-}\nu_{\tau}\bar{\nu}_{\mu}$. The measured values are $\mathcal{R}(D^{*})=0.281\pm0.018\pm0.024$ and $\mathcal{R}(D^{0})=0.441\pm0.060\pm0.066$, where the first uncertainty is statistical and the second is systematic. The correlation between these measurements is $\rho=-0.43$. Results are consistent with the current average of these quantities and are at a combined 1.9 standard deviations from the predictions based on lepton flavor universality in the Standard Model

Measurement of Ξc+\Xi_{c}^{+} production in ppPb collisions at sNN=8.16\sqrt{s_{NN}}=8.16 TeV at LHCb

International audienceA study of prompt $\Xi_{c}^{+}$ production in proton-lead collisions is performed with the LHCb experiment at a centre-of-mass energy per nucleon pair of 8.16 TeV in 2016 in $p$Pb and Pb$p$ collisions with an estimated integrated luminosity of approximately 12.5 and 17.4 nb$^{-1}$, respectively. The $\Xi_{c}^{+}$ production cross-section, as well as the $\Xi_{c}^{+}$ to $\Lambda_{c}^{+}$ production cross-section ratio, are measured as a function of the transverse momentum and rapidity and compared to latest theory predictions. The forward-backward asymmetry is also measured as a function of the $\Xi_{c}^{+}$ transverse momentum