Cell-based high-content approach for SARS-CoV-2 neutralization identifies unique monoclonal antibodies and PI3K pathway inhibitors.

The sudden rise of the SARS-CoV-2 virus and the delay in development of effective therapeutics for mitigation made evident a need for ways to screen compounds that can block infection and prevent further pathogenesis and spread. However, identifying effective drugs that are efficacious against viral infection and replication with minimal toxicity for the patient can be difficult. Monoclonal antibodies were shown to be effective, but as the SARS-CoV-2 mutated, these antibodies became ineffective. Small-molecule antivirals were identified using pseudovirus constructs to recapitulate infection in nonhuman cells, such as Vero E6 cells. However, the impact was limited due to poor translation of these compounds in the clinical setting. This is partly due to the lack of similarity of screening platforms to the in vivo physiology of the patient and partly because drugs effective in vitro showed dose-limiting toxicities. In this study, we performed two high-throughput screens in human lung adenocarcinoma cells with authentic SARS-CoV-2 virus to identify both monoclonal antibodies that neutralize the virus and clinically useful kinase inhibitors to block the virus and prioritize minimal host toxicity. Using high-content imaging combined with single-cell and multidimensional analysis, we identified antibodies and kinase inhibitors that reduce viral infection without affecting the host. Our screening technique uncovered novel antibodies and overlooked kinase inhibitors (i.e., PIK3i, mTORi, and multiple RTKi) that could be effective against the SARS-CoV-2 virus. Further characterization of these molecules will streamline the repurposing of compounds for the treatment of future pandemics and uncover novel mechanisms viruses use to hijack and infect host cells

Rendimento de carcaça de três grupos genéticos de frangos de corte alimentados com rações contendo diferentes teores de proteína

Este experimento foi realizado com o objetivo de avaliar o rendimento de carcaça de três grupos genéticos de frangos de corte, produzidos na UFV, denominados UFV1, UFV2 e UFV3, alimentados com ração única na fase inicial e rações isoenergéticas contendo 16,5; 18,0; 19,5; e 21,0% de proteína bruta (PB) na fase final. Duzentos e quarenta pintos de cada grupo genético, no total de 720 pintos, foram alojados em 48 boxes até 42 dias de idade. No 43º dia, dois machos e duas fêmeas, de cada box, foram pesados e abatidos para determinação do rendimento de carcaça. O delineamento experimental usado foi o inteiramente casualizado em arranjo fatorial 3 x 4 x 2 (grupo genético, nível de proteína e sexo), com quatro repetições, num total de 192 aves. Os valores dos dados de duas aves por sexo foram usados para o cálculo da média. Houve diferença entre grupos genéticos para rendimentos de carcaça, peito e coxa, e o grupo UFV1 apresentou os melhores resultados. Entre sexo, os machos foram superiores nos rendimentos de coxa e sobre-coxa, porém observaram-se maiores rendimentos de peito e gordura abdominal para as fêmeas. À medida que o nível de PB da ração aumentou, o rendimento de carcaça e gordura abdominal reduziu e o de coxa aumentou (efeito linear). Não foi verificado efeito significativo de interação simples dos fatores grupo genético, níveis de proteína e sexo.The objective of this experiment was to evaluate the carcass yield of three genetic groups of broilers chickens, obtained at UFV, called UFV1, UFV2 and UFV3. The chicks were fed an unique diet during the initial phase period and isocaloric diet containing 16.5, 18.0, 19.5, and 21.0% of crude protein (CP) at the finishing phase. Two hundred and forty chicks of each genetic group, totaling 720 chicks, were allotted to 48 floor pens up to 42 days of age. At43rd day, two males and two females of each pen were weighed and slaughtered for the determination of the carcass yield. A completely randomized experimental design was used in a 3x4x2 factorial arrangement (genetic group, protein level and sex) with four replications, totaling 192 chicks. The values of two chickens data per sex were used in the mean calculation. There were differences among genetic groups for yields of carcass, breast and thigh, and UFV1 group showed the best results. Males had higher yields of thigh and drumstick, however, higher breast and abdominal fat yields were observed for females. As dietary CP levels increased, carcass yield and abdominal fat reduced and thigh yield increased (linear effect). No significant effect was observed for single interaction involving genetic group, protein levels and sex

Cell-free DNA profiling of metastatic prostate cancer reveals microsatellite instability, structural rearrangements and clonal hematopoiesis.

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.BACKGROUND: There are multiple existing and emerging therapeutic avenues for metastatic prostate cancer, with a common denominator, which is the need for predictive biomarkers. Circulating tumor DNA (ctDNA) has the potential to cost-efficiently accelerate precision medicine trials to improve clinical efficacy and diminish costs and toxicity. However, comprehensive ctDNA profiling in metastatic prostate cancer to date has been limited. METHODS: A combination of targeted and low-pass whole genome sequencing was performed on plasma cell-free DNA and matched white blood cell germline DNA in 364 blood samples from 217 metastatic prostate cancer patients. RESULTS: ctDNA was detected in 85.9% of baseline samples, correlated to line of therapy and was mirrored by circulating tumor cell enumeration of synchronous blood samples. Comprehensive profiling of the androgen receptor (AR) revealed a continuous increase in the fraction of patients with intra-AR structural variation, from 15.4% during first-line metastatic castration-resistant prostate cancer therapy to 45.2% in fourth line, indicating a continuous evolution of AR during the course of the disease. Patients displayed frequent alterations in DNA repair deficiency genes (18.0%). Additionally, the microsatellite instability phenotype was identified in 3.81% of eligible samples (≥ 0.1 ctDNA fraction). Sequencing of non-repetitive intronic and exonic regions of PTEN, RB1, and TP53 detected biallelic inactivation in 47.5%, 20.3%, and 44.1% of samples with ≥ 0.2 ctDNA fraction, respectively. Only one patient carried a clonal high-impact variant without a detectable second hit. Intronic high-impact structural variation was twice as common as exonic mutations in PTEN and RB1. Finally, 14.6% of patients presented false positive variants due to clonal hematopoiesis, commonly ignored in commercially available assays. CONCLUSIONS: ctDNA profiles appear to mirror the genomic landscape of metastatic prostate cancer tissue and may cost-efficiently provide somatic information in clinical trials designed to identify predictive biomarkers. However, intronic sequencing of the interrogated tumor suppressors challenges the ubiquitous focus on coding regions and is vital, together with profiling of synchronous white blood cells, to minimize erroneous assignments which in turn may confound results and impede true associations in clinical trials.The Belgian Foundation Against Cancer (grant number C/2014/227); Kom op tegen Kanker (Stand up to Cancer), the Flemish Cancer Society (grant number 00000000116000000206); Royal College of Surgeons/Cancer Research UK (C19198/A1533); The Cancer Research Funds of Radiumhemmet, through the PCM program at KI (grant number 163012); The Erling-Persson family foundation (grant number 4-2689-2016); the Swedish Research Council (grant number K2010-70X-20430-04-3), and the Swedish Cancer Foundation (grant number 09-0677)