Adaphostin toxicity in a sensitive non-small cell lung cancer model is mediated through Nrf2 signaling and heme oxygenase 1

<p>Abstract</p> <p>Background</p> <p>Preclinical toxicity of adaphostin has been related to oxidative stress. This study investigated the regulatory mechanism underlying adaphostin induction of heme oxygenase 1 (HMOX1) which plays a significant role in modulation of drug-induced toxicity in the non-small cell lung cancer cell line model, NCI-H522.</p> <p>Methods</p> <p>The transcriptional response of NCI-H522 to adaphostin prominently involved oxidative stress genes, particularly HMOX1. Reactive oxygen species (ROS) involvement was additionally established by generation of ROS prior to modulation of adaphostin-toxicity with antioxidants. To identify up-stream regulatory elements of HMOX1, immunofluorescence was used to evaluate nuclear translocation of the transcription factor, NF-E2-related factor 2 (Nrf2), in the presence of adaphostin. The PI3-kinase inhibitor, wortmannin, was employed as a pharmacological inhibitor of this process.</p> <p>Results</p> <p>Generation of ROS provided a substantial foundation for the sensitivity of NCI-H522 to adaphostin. However, in contrast to leukemia cell lines, transcriptional response to oxidative stress was associated with induction of HMOX1, which was dependent on nuclear translocation of the transcription factor, Nrf2. Pretreatment of cells with wortmannin inhibited translocation of Nrf2 and induction of HMOX1. Wortmannin pretreatment was also able to diminish adaphostin induction of HMOX1, and as a consequence, enhance the toxicity of adaphostin to NCI-H522.</p> <p>Conclusions</p> <p>Adaphostin-induced oxidative stress in NCI-H522 was mediated through nuclear translocation of Nrf2 leading to upregulation of HMOX1. Inhibition of Nrf2 translocation by wortmannin inhibited this cytoprotective response, and enhanced the toxicity of adaphostin, suggesting that inhibitors of the PI3K pathway, such as wortmannin, might augment the antiproliferative effects of adaphostin in solid tumors that depend on the Nrf2/ARE pathway for protection against oxidative stress.</p

Data Availability Principles and Practice

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Evaluation of the selectivity and sensitivity of isoform- and mutation-specific RAS antibodies

Researchers rely largely on antibodies to measure the abundance, activity, and localization of a protein, information that provides critical insight into both normal and pathological cellular functions. However, antibodies are not always reliable or universally valid for the methods in which they are used; in particular, the reliability of commercial antibodies against RAS is highly variable. Waters et al . rigorously assessed 22 commercially available RAS antibodies for their utility to detect the distinct RAS isoforms in various cell types and for their use in specific analytical methods. Their findings show how reliably one can interpret the data acquired from each reagent

Classical RAS proteins are not essential for paradoxical ERK activation induced by RAF inhibitors.

RAF inhibitors unexpectedly induce ERK signaling in normal and tumor cells with elevated RAS activity. Paradoxical activation is believed to be RAS dependent. In this study, we showed that LY3009120, a pan-RAF inhibitor, can unexpectedly cause paradoxical ERK activation in KRASG12C-dependent lung cancer cell lines, when KRAS is inhibited by ARS1620, a KRASG12C inhibitor. Using H/N/KRAS-less mouse embryonic fibroblasts, we discovered that classical RAS proteins are not essential for RAF inhibitor-induced paradoxical ERK signaling. In their absence, RAF inhibitors can induce ERK phosphorylation, ERK target gene transcription, and cell proliferation. We further showed that the MRAS/SHOC2 complex is required for this process. This study highlights the complexity of the allosteric RAF regulation by RAF inhibitors, and the importance of other RAS-related proteins in this process

Turbulent diapycnal fluxes as a pilot Essential Ocean Variable

Peer reviewed: TrueAcknowledgements: All co-authors, and especially the lead authors acknowledge the tremendous input from J. Moum who helped re-oriented the main topic of this white paper toward what the ocean community is now considering the best option for turbulence data to become an EOV. We are also in debt of all the pioneers who participated in the development of today’s turbulence technology.We contend that ocean turbulent fluxes should be included in the list of Essential Ocean Variables (EOVs) created by the Global Ocean Observing System. This list aims to identify variables that are essential to observe to inform policy and maintain a healthy and resilient ocean. Diapycnal turbulent fluxes quantify the rates of exchange of tracers (such as temperature, salinity, density or nutrients, all of which are already EOVs) across a density layer. Measuring them is necessary to close the tracer concentration budgets of these quantities. Measuring turbulent fluxes of buoyancy (Jb ), heat (Jq ), salinity (JS ) or any other tracer requires either synchronous microscale (a few centimeters) measurements of both the vector velocity and the scalar (e.g., temperature) to produce time series of the highly correlated perturbations of the two variables, or microscale measurements of turbulent dissipation rates of kinetic energy (ϵ) and of thermal/salinity/tracer variance (χ), from which fluxes can be derived. Unlike isopycnal turbulent fluxes, which are dominated by the mesoscale (tens of kilometers), microscale diapycnal fluxes cannot be derived as the product of existing EOVs, but rather require observations at the appropriate scales. The instrumentation, standardization of measurement practices, and data coordination of turbulence observations have advanced greatly in the past decade and are becoming increasingly robust. With more routine measurements, we can begin to unravel the relationships between physical mixing processes and ecosystem health. In addition to laying out the scientific relevance of the turbulent diapycnal fluxes, this review also compiles the current developments steering the community toward such routine measurements, strengthening the case for registering the turbulent diapycnal fluxes as an pilot Essential Ocean Variable

Corrigendum: Turbulent diapycnal fluxes as a pilot Essential Ocean Variable

Peer reviewed: Tru