The role of APOBEC3B in lung tumor evolution and targeted cancer therapy resistance

In this study, the impact of the apolipoprotein B mRNA-editing catalytic subunit-like (APOBEC) enzyme APOBEC3B (A3B) on epidermal growth factor receptor (EGFR)-driven lung cancer was assessed. A3B expression in EGFR mutant (EGFRmut) non-small-cell lung cancer (NSCLC) mouse models constrained tumorigenesis, while A3B expression in tumors treated with EGFR-targeted cancer therapy was associated with treatment resistance. Analyses of human NSCLC models treated with EGFR-targeted therapy showed upregulation of A3B and revealed therapy-induced activation of nuclear factor kappa B (NF-κB) as an inducer of A3B expression. Significantly reduced viability was observed with A3B deficiency, and A3B was required for the enrichment of APOBEC mutation signatures, in targeted therapy-treated human NSCLC preclinical models. Upregulation of A3B was confirmed in patients with NSCLC treated with EGFR-targeted therapy. This study uncovers the multifaceted roles of A3B in NSCLC and identifies A3B as a potential target for more durable responses to targeted cancer therapy.</p

Albiglutide and cardiovascular outcomes in patients with type 2 diabetes and cardiovascular disease (Harmony Outcomes): a double-blind, randomised placebo-controlled trial

Background: Glucagon-like peptide 1 receptor agonists differ in chemical structure, duration of action, and in their effects on clinical outcomes. The cardiovascular effects of once-weekly albiglutide in type 2 diabetes are unknown. We aimed to determine the safety and efficacy of albiglutide in preventing cardiovascular death, myocardial infarction, or stroke. Methods: We did a double-blind, randomised, placebo-controlled trial in 610 sites across 28 countries. We randomly assigned patients aged 40 years and older with type 2 diabetes and cardiovascular disease (at a 1:1 ratio) to groups that either received a subcutaneous injection of albiglutide (30–50 mg, based on glycaemic response and tolerability) or of a matched volume of placebo once a week, in addition to their standard care. Investigators used an interactive voice or web response system to obtain treatment assignment, and patients and all study investigators were masked to their treatment allocation. We hypothesised that albiglutide would be non-inferior to placebo for the primary outcome of the first occurrence of cardiovascular death, myocardial infarction, or stroke, which was assessed in the intention-to-treat population. If non-inferiority was confirmed by an upper limit of the 95% CI for a hazard ratio of less than 1·30, closed testing for superiority was prespecified. This study is registered with ClinicalTrials.gov, number NCT02465515. Findings: Patients were screened between July 1, 2015, and Nov 24, 2016. 10 793 patients were screened and 9463 participants were enrolled and randomly assigned to groups: 4731 patients were assigned to receive albiglutide and 4732 patients to receive placebo. On Nov 8, 2017, it was determined that 611 primary endpoints and a median follow-up of at least 1·5 years had accrued, and participants returned for a final visit and discontinuation from study treatment; the last patient visit was on March 12, 2018. These 9463 patients, the intention-to-treat population, were evaluated for a median duration of 1·6 years and were assessed for the primary outcome. The primary composite outcome occurred in 338 (7%) of 4731 patients at an incidence rate of 4·6 events per 100 person-years in the albiglutide group and in 428 (9%) of 4732 patients at an incidence rate of 5·9 events per 100 person-years in the placebo group (hazard ratio 0·78, 95% CI 0·68–0·90), which indicated that albiglutide was superior to placebo (p&lt;0·0001 for non-inferiority; p=0·0006 for superiority). The incidence of acute pancreatitis (ten patients in the albiglutide group and seven patients in the placebo group), pancreatic cancer (six patients in the albiglutide group and five patients in the placebo group), medullary thyroid carcinoma (zero patients in both groups), and other serious adverse events did not differ between the two groups. There were three (&lt;1%) deaths in the placebo group that were assessed by investigators, who were masked to study drug assignment, to be treatment-related and two (&lt;1%) deaths in the albiglutide group. Interpretation: In patients with type 2 diabetes and cardiovascular disease, albiglutide was superior to placebo with respect to major adverse cardiovascular events. Evidence-based glucagon-like peptide 1 receptor agonists should therefore be considered as part of a comprehensive strategy to reduce the risk of cardiovascular events in patients with type 2 diabetes. Funding: GlaxoSmithKline

Nesting Behavior of Acapulco damselfish (Stegastes acapulcoensis): pairs defend nests more vigorously than solitary males

Unique pair behavior was observed in Stegastes acapulcoensis, the Acapulco damselfish, in the reefs of Santa Elena Bay of Costa Rica. Observations and video were collected on aggression and territoriality demonstrated by both male-female pairs and solitary males guarding eggs. Agonistic chases and chase distance were recorded over four days during snorkeling expeditions. Results compared parental care behavior (pairs versus solitary) with aggressive chases and chase distance. Solitary male Acapulco damselfish chase significantly less times than paired. Additionally, the paired males chase significantly farther than solitary. Males were observed to defend territory and eggs using aggressive chases, while paired females remained within the nest. These data and behavioral differences may suggest that pair behavior is an effective defensive strategy, likely motivated by a common goal of ensuring survival of offspring. Furthermore, pair defense may be influenced by differences in parental investment and contribution to reproductive success. Un comportamiento único en pareja de Stegastes acapulcoensis, el pez damisela de Acapulco, se observó en los arrecifes de la Bahía de Santa Elena de Costa Rica. Se tomaron observaciones y videos del comportamiento agresivo y territorialidad demostrados por parejas de machos-hembras y también machos solitarios cuidando masas de huevos. Las persecuciones agonísticas y la distancia de persecución en el mar se registraron durante cuatro días de buceo. Los resultados compararon el comportamiento de cuidado parental (parejas contra machos solitarios) con respecto a persecuciones agresivas y distancia de persecución. Los machos solitarios persiguieron significativamente menos veces que los machos emparejados. Además, los machos emparejados persiguieron significativamente más lejos que los machos solitarios. Se observó que los machos defendían el territorio y los huevos usando persecuciones agresivas, mientras que sus parejas hembras quedaban cerca del nido. Estos datos y las diferencias de comportamiento demuestran que el comportamiento de las parejas es una estrategia defensiva, y es probable que sea motivada por un objetivo común de asegurar la sobrevivencia de la progenie. Finalmente, la defensa proporcionada por las parejas puede estar influenciada por el costo de cuidar sus huevos y la contribución positiva al éxito reproductivo.https://digitalcommons.usf.edu/tropical_ecology/1427/thumbnail.jp

Nesting Behavior of Acapulco damselfish (Stegastes acapulcoensis): pairs defend nests more vigorously than solitary males

Unique pair behavior was observed in Stegastes acapulcoensis, the Acapulco damselfish, in the reefs of Santa Elena Bay of Costa Rica. Observations and video were collected on aggression and territoriality demonstrated by both male-female pairs and solitary males guarding eggs. Agonistic chases and chase distance were recorded over four days during snorkeling expeditions. Results compared parental care behavior (pairs versus solitary) with aggressive chases and chase distance. Solitary male Acapulco damselfish chase significantly less times than paired. Additionally, the paired males chase significantly farther than solitary. Males were observed to defend territory and eggs using aggressive chases, while paired females remained within the nest. These data and behavioral differences may suggest that pair behavior is an effective defensive strategy, likely motivated by a common goal of ensuring survival of offspring. Furthermore, pair defense may be influenced by differences in parental investment and contribution to reproductive success. Un comportamiento único en pareja de Stegastes acapulcoensis, el pez damisela de Acapulco, se observó en los arrecifes de la Bahía de Santa Elena de Costa Rica. Se tomaron observaciones y videos del comportamiento agresivo y territorialidad demostrados por parejas de machos-hembras y también machos solitarios cuidando masas de huevos. Las persecuciones agonísticas y la distancia de persecución en el mar se registraron durante cuatro días de buceo. Los resultados compararon el comportamiento de cuidado parental (parejas contra machos solitarios) con respecto a persecuciones agresivas y distancia de persecución. Los machos solitarios persiguieron significativamente menos veces que los machos emparejados. Además, los machos emparejados persiguieron significativamente más lejos que los machos solitarios. Se observó que los machos defendían el territorio y los huevos usando persecuciones agresivas, mientras que sus parejas hembras quedaban cerca del nido. Estos datos y las diferencias de comportamiento demuestran que el comportamiento de las parejas es una estrategia defensiva, y es probable que sea motivada por un objetivo común de asegurar la sobrevivencia de la progenie. Finalmente, la defensa proporcionada por las parejas puede estar influenciada por el costo de cuidar sus huevos y la contribución positiva al éxito reproductivo.https://digitalcommons.usf.edu/tropical_ecology/1427/thumbnail.jp

Deep learning-based characterization of the drug tolerant persister cell state in lung cancer.

141 Background: Lung cancer is the leading cause of cancer-related lethality globally. Targeted therapies improve the clinical outcome of cancer treatment; however, a subpopulation of cancer cells survive during initial therapy and evolve into drug tolerant persister cells (DTPs) that maintain a residual disease reservoir. Residual disease preludes acquired resistance and tumor progression; therefore, identifying and eliminating DTPs could benefit future treatment paradigms. We have shown that Hippo pathway effector YAP1 (Yes Associated Protein-1) is activated in oncogene-driven lung cancers when cancer cells are exposed to various targeted therapies, such as EGFR, ALK, and RAS inhibitors. YAP1 transcriptional activation during targeted therapy is characterized by its increased nuclear localization and interaction with transcription factors. This activation promotes the expression of genes involved in cell survival, cellular plasticity, and metabolic reprogramming at the residual disease state. We hypothesized that detection of intrinsic or acquired persister cells may aid the development of optimized treatment regimens. In this study, we are building image-based deep learning models to identify YAP1 activation-mediated DTPs from histologically stained slides. Methods: H&amp;E or YAP-stained immunohistochemistry (IHC) images from clinical lung cancer and patient-derived tumor xenograft samples were collected throughout targeted therapy and were annotated with semi-automation using high performance computing clusters. A deep learning model (U-Net algorithm) was used for image segmentation, training, validation, and testing. Results: 1638 images were annotated and over 80,000 patches from these images for YAP positive cells (or regions of interest) comprised the training dataset with semi-supervised automation. Subsequently, we built a customized deep-learning model to detect YAP-mediated DTP cell states from whole histopathological image slides. The deep learning-based model achieved excellent accuracy of 0.8238 and 0.9091 in training, and 0.8040 and 0.8949 in validation datasets for two different annotations, respectively. For a test dataset, the model obtained 0.81 and 0.902 accuracy for the two annotations, respectively. Conclusions: We have constructed a deep learning convolutional neural network model to infer the presence of the YAP1 activation-mediated drug tolerant persister cell state prior to or during targeted treatment of lung cancer. Implementing our AI-based model into routine lung cancer care in the future could identify patient subpopulations with YAP1 activated tumors who would most benefit from receiving YAP1-targeted small molecule inhibitors

Modulation of <i>in Vitro</i> SARS-CoV‑2 Infection by <i>Stephania tetrandra</i> and Its Alkaloid Constituents

Botanical natural products have been widely consumed for their purported usefulness against COVID-19. Here, six botanical species from multiple sources and 173 isolated natural product compounds were screened for blockade of wild-type (WT) SARS-CoV-2 infection in human 293T epithelial cells overexpressing ACE-2 and TMPRSS2 protease (293TAT). Antiviral activity was demonstrated by an extract from Stephania tetrandra. Extract fractionation, liquid chromatography–mass spectrometry (LC-MS), antiviral assays, and computational analyses revealed that the alkaloid fraction and purified alkaloids tetrandrine, fangchinoline, and cepharanthine inhibited WT SARS-CoV-2 infection. The alkaloids and alkaloid fraction also inhibited the delta variant of concern but not WT SARS-CoV-2 in VeroAT cells. Membrane permeability assays demonstrate that the alkaloids are biologically available, although fangchinoline showed lower permeability than tetrandrine. At high concentrations, the extract, alkaloid fractions, and pure alkaloids induced phospholipidosis in 293TAT cells and less so in VeroAT cells. Gene expression profiling during virus infection suggested that alkaloid fraction and tetrandrine displayed similar effects on cellular gene expression and pathways, while fangchinoline showed distinct effects on cells. Our study demonstrates a multifaceted approach to systematically investigate the diverse activities conferred by complex botanical mixtures, their cell-context specificity, and their pleiotropic effects on biological systems

Modulation of <i>in Vitro</i> SARS-CoV‑2 Infection by <i>Stephania tetrandra</i> and Its Alkaloid Constituents

Botanical natural products have been widely consumed for their purported usefulness against COVID-19. Here, six botanical species from multiple sources and 173 isolated natural product compounds were screened for blockade of wild-type (WT) SARS-CoV-2 infection in human 293T epithelial cells overexpressing ACE-2 and TMPRSS2 protease (293TAT). Antiviral activity was demonstrated by an extract from Stephania tetrandra. Extract fractionation, liquid chromatography–mass spectrometry (LC-MS), antiviral assays, and computational analyses revealed that the alkaloid fraction and purified alkaloids tetrandrine, fangchinoline, and cepharanthine inhibited WT SARS-CoV-2 infection. The alkaloids and alkaloid fraction also inhibited the delta variant of concern but not WT SARS-CoV-2 in VeroAT cells. Membrane permeability assays demonstrate that the alkaloids are biologically available, although fangchinoline showed lower permeability than tetrandrine. At high concentrations, the extract, alkaloid fractions, and pure alkaloids induced phospholipidosis in 293TAT cells and less so in VeroAT cells. Gene expression profiling during virus infection suggested that alkaloid fraction and tetrandrine displayed similar effects on cellular gene expression and pathways, while fangchinoline showed distinct effects on cells. Our study demonstrates a multifaceted approach to systematically investigate the diverse activities conferred by complex botanical mixtures, their cell-context specificity, and their pleiotropic effects on biological systems

Modulation of <i>in Vitro</i> SARS-CoV‑2 Infection by <i>Stephania tetrandra</i> and Its Alkaloid Constituents

Botanical natural products have been widely consumed for their purported usefulness against COVID-19. Here, six botanical species from multiple sources and 173 isolated natural product compounds were screened for blockade of wild-type (WT) SARS-CoV-2 infection in human 293T epithelial cells overexpressing ACE-2 and TMPRSS2 protease (293TAT). Antiviral activity was demonstrated by an extract from Stephania tetrandra. Extract fractionation, liquid chromatography–mass spectrometry (LC-MS), antiviral assays, and computational analyses revealed that the alkaloid fraction and purified alkaloids tetrandrine, fangchinoline, and cepharanthine inhibited WT SARS-CoV-2 infection. The alkaloids and alkaloid fraction also inhibited the delta variant of concern but not WT SARS-CoV-2 in VeroAT cells. Membrane permeability assays demonstrate that the alkaloids are biologically available, although fangchinoline showed lower permeability than tetrandrine. At high concentrations, the extract, alkaloid fractions, and pure alkaloids induced phospholipidosis in 293TAT cells and less so in VeroAT cells. Gene expression profiling during virus infection suggested that alkaloid fraction and tetrandrine displayed similar effects on cellular gene expression and pathways, while fangchinoline showed distinct effects on cells. Our study demonstrates a multifaceted approach to systematically investigate the diverse activities conferred by complex botanical mixtures, their cell-context specificity, and their pleiotropic effects on biological systems

Modulation of <i>in Vitro</i> SARS-CoV‑2 Infection by <i>Stephania tetrandra</i> and Its Alkaloid Constituents

Botanical natural products have been widely consumed for their purported usefulness against COVID-19. Here, six botanical species from multiple sources and 173 isolated natural product compounds were screened for blockade of wild-type (WT) SARS-CoV-2 infection in human 293T epithelial cells overexpressing ACE-2 and TMPRSS2 protease (293TAT). Antiviral activity was demonstrated by an extract from Stephania tetrandra. Extract fractionation, liquid chromatography–mass spectrometry (LC-MS), antiviral assays, and computational analyses revealed that the alkaloid fraction and purified alkaloids tetrandrine, fangchinoline, and cepharanthine inhibited WT SARS-CoV-2 infection. The alkaloids and alkaloid fraction also inhibited the delta variant of concern but not WT SARS-CoV-2 in VeroAT cells. Membrane permeability assays demonstrate that the alkaloids are biologically available, although fangchinoline showed lower permeability than tetrandrine. At high concentrations, the extract, alkaloid fractions, and pure alkaloids induced phospholipidosis in 293TAT cells and less so in VeroAT cells. Gene expression profiling during virus infection suggested that alkaloid fraction and tetrandrine displayed similar effects on cellular gene expression and pathways, while fangchinoline showed distinct effects on cells. Our study demonstrates a multifaceted approach to systematically investigate the diverse activities conferred by complex botanical mixtures, their cell-context specificity, and their pleiotropic effects on biological systems