Characterization of Vemurafenib Phototoxicity in a Mouse Model

Vemurafenib is a first-in-class, small molecule B-Raf kinase inhibitor for the treatment of patients with unresectable or metastatic melanoma carrying the BRAFV600E mutation, commercially available since 2011. A general phototoxic potential was identified early during development; however, based on results of an animal study in hairless rats, it was concluded that there would exist no relevant risk for humans. Surprisingly, signs of clinical photosensitivity were reported in many patients during clinical development. Therefore, it became a fundamental question to understand this discrepancy. An established mouse model (oral UV-Local Lymph Node Assay, UV-LLNA) for the assessment of in vivo photosafety was used to investigate the impact of formulations, dose levels, duration of treatment, and timing of irradiation. Moreover, a basic pharmacokinetic profile was established within the same mouse strain. We were able to demonstrate dose- and time-dependent phototoxicity of vemurafenib using commercially available tablets (stabilized amorphous material). The lowest phototoxic dose was 350mg/kg administrated for 3 consecutive days followed by exposure to UV-visible irradiation at a UVA-normalized dose of 10 J/cm2. In comparison, pure vemurafenib, which easily forms crystalline variants and is known to have poor bioavailability, was tested at 350mg/kg, and no signs of phototoxicity could be seen. The most apparent difference between the early study in hairless rats and this study in mice was the spectral range of the irradiation light source (350-400nm vs 320-700nm). Because vemurafenib does not absorb sufficiently light above 350nm, this difference can easily explain the negative earlier study result in hairless rat

Non-photochemical quenching estimates from in situ spectroradiometer measurements: implications on remote sensing of sun-induced chlorophyll fluorescence in lakes

Quantum yield of fluorescence (ϕF) is key to interpret remote measurements of sun-induced fluorescence (SIF), and whether the SIF signal is governed by photochemical quenching (PQ) or non-photochemical quenching (NPQ). Disentangling PQ from NPQ allows using SIF estimates in various applications in aquatic optics. However, obtaining ϕF is challenging due to its high temporal and physiological variability, and the combined measurements needed to enclose all relevant optical paths. In inland waters, this type of data is scarce and information on diurnal and seasonal ϕF dynamics are almost unknown. Using an autonomous hyperspectral Thetis profiler in Lake Geneva, we demonstrate how to estimate ϕF using an ensemble of in-situ measurements acquired between 2018 to 2021. We use vertical and temporal changes in retrieved ϕF to determine NPQ and PQ conditions. We observed NPQ in 36% of the total daytime profiles used in the ϕF analysis. While downwelling irradiance is a significant contributor to ϕF, its role cannot be easily interpreted. Other factors such as phytoplankton photoregulation and assemblages also likely play significant roles in quenching mechanisms. We conclude that an adapted approach exploiting in-situ data is suitable to determine diurnal and seasonal NPQ occurrence, and helps develop future remote sensing algorithms

The imprint of primary production on high-frequency profiles of lake optical properties

Water inherent optical properties (IOPs) contain integrative information on the optical constituents of surface waters. In lakes, IOP measurements have not been traditionally collected. This study describes how high-frequency IOP profiles can be used to document short-term physical and biogeochemical processes that ultimately influence the long-term trajectory of lake ecosystems. Between October 2018 and May 2020, we collected 1373 high-resolution hyperspectral IOP profiles in the uppermost 50 m of the large mesotrophic Lake Geneva (Switzerland-France), using an autonomous profiler. A data set of this size and content does not exist for any other lake. Results showed seasonal variations in the IOPs, following the expected dynamic of phytoplankton. We found systematic diel patterns in the IOPs. Phases of these diel cycles were consistent year-round, and amplitudes correlated to the diurnal variations of dissolved oxygen, clarifying the link between IOPs and phytoplankton metabolism. Diel amplitudes were largest in spring and summer under low wind condition. Wind-driven changes in thermal stratification impacted the dynamic of the IOPs, illustrating the potential of high-frequency profiles of water optical properties to increase our understanding of carbon cycling in lake ecosystems

Oncostatin M suppresses browning of white adipocytes via gp130-STAT3 signaling

Objective Obesity is associated with low-grade adipose tissue inflammation and locally elevated levels of several glycoprotein 130 (gp130) cytokines. The conversion of white into brown-like adipocytes (browning) may increase energy expenditure and revert the positive energy balance that underlies obesity. Although different gp130 cytokines and their downstream targets were shown to regulate expression of the key browning marker uncoupling protein 1 (Ucp1), it remains largely unknown how this contributes to the development and maintenance of obesity. Herein, we aim to study the role of gp130 cytokine signaling in white adipose tissue (WAT) browning in the obese state. Methods Protein and gene expression levels of UCP1 and other thermogenic markers were assessed in a subcutaneous adipocyte cell line, adipose tissue depots from control or adipocyte-specific gp130 knockout (gp130Δadipo) mice fed either chow or a high-fat diet (HFD), or subcutaneous WAT biopsies from a human cohort of lean and obese subjects. WAT browning was modelled in vitro by exposing mature adipocytes to isoproterenol subsequent to stimulation with gp130 cytokines. ERK and JAK-STAT signaling were blocked using the inhibitors U0126 and Tofacitinib, respectively. Results Inguinal WAT of HFD-fed gp130Δadipo mice exhibited significantly elevated levels of UCP1 and other browning markers such as Cidea and Pgc-1α. In vitro, treatment with the gp130 cytokine oncostatin M (OSM) lowered isoproterenol-induced UCP1 protein and gene expression levels in a dose-dependent manner. Mechanistically, OSM mediated the inhibition of Ucp1 via the JAK-STAT but not the ERK pathway. In line with mouse data, OSM gene expression in human WAT positively correlated with BMI (r=0.284, p=0.021, n=66) and negatively with UCP1 expression (r=-0.413, p<0.001, n=66). Conclusions Our data support the notion that OSM negatively regulates thermogenesis in WAT and, thus, may be an attractive target to treat obesity

Protective T Cell–Independent Antiviral Antibody Responses Are Dependent on Complement

Complement is part of the innate immune system and one of the first lines of host defense against infections. Its importance was evaluated in this study in virus infections in mice deficient either in soluble complement factors (C3−/−, C4−/−) or in the complement signaling complex (complement receptor [CR]2−/−, CD19−/−). The induction of the initial T cell–independent neutralizing immunoglobulin (Ig)M antibody response to vesicular stomatitis virus (VSV), poliomyelitis virus, and recombinant vaccinia virus depended on efficient antigen trapping by CR3 and -4–expressing macrophages of the splenic marginal zone. Neutralizing IgM and IgG antibody responses were largely independent of CR2-mediated stimulation of B cells when mice were infected with live virus. In contrast, immunizations with nonreplicating antigens revealed an important role of B cell stimulation via CR2 in the switch to IgG. The complement cascade was activated after infection with VSV via the classical pathway, and active complement cleavage products augmented the effector function of neutralizing IgM and IgG antibodies to VSV by a factor of 10–100. Absence of the early neutralizing antibody responses, together with the reduced efficiency of neutralizing IgM in C3−/− mice, led to a drastically enhanced susceptibility to disease after infection with VSV

Long-Term Spatiotemporal Variability of Whitings in Lake Geneva from Multispectral Remote Sensing and Machine Learning

Whiting events are massive calcite precipitation events turning hardwater lake waters to a milky turquoise color. Herein, we use a multispectral remote sensing approach to describe the spatial and temporal occurrences of whitings in Lake Geneva from 2013 to 2021. Landsat-8, Sentinel-2, and Sentinel-3 sensors are combined to derive the AreaBGR index and identify whitings using appropriate filters. 95% of the detected whitings are located in the northeastern part of the lake and occur in a highly reproducible environmental setting. An extended time series of whitings in the last 60 years is reconstructed from a random forest algorithm and analyzed through a Bayesian decomposition for annual and seasonal trends. The annual number of whiting days between 1958 and 2021 does not follow any particular monotonic trend. The inter-annual changes of whiting occurrences significantly correlate to the Western Mediterranean Oscillation Index. Spring whitings have increased since 2000 and significantly follow the Atlantic Multidecadal Oscillation index. Future climate change in the Mediterranean Sea and the Atlantic Ocean could induce more variable and earlier whiting events in Lake Geneva

Developing and testing a Corona VaccinE tRiAL pLatform (COVERALL) to study Covid-19 vaccine response in immunocompromised patients

BACKGROUND The rapid course of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic calls for fast implementation of clinical trials to assess the effects of new treatment and prophylactic interventions. Building trial platforms embedded in existing data infrastructures is an ideal way to address such questions within well-defined subpopulations. METHODS We developed a trial platform building on the infrastructure of two established national cohort studies: the Swiss human immunodeficiency virus (HIV) Cohort Study (SHCS) and Swiss Transplant Cohort Study (STCS). In a pilot trial, termed Corona VaccinE tRiAL pLatform (COVERALL), we assessed the vaccine efficacy of the first two licensed SARS-CoV-2 vaccines in Switzerland and the functionality of the trial platform. RESULTS Using Research Electronic Data Capture (REDCap), we developed a trial platform integrating the infrastructure of the SHCS and STCS. An algorithm identifying eligible patients, as well as baseline data transfer ensured a fast inclusion procedure for eligible patients. We implemented convenient re-directions between the different data entry systems to ensure intuitive data entry for the participating study personnel. The trial platform, including a randomization algorithm ensuring balance among different subgroups, was continuously adapted to changing guidelines concerning vaccination policies. We were able to randomize and vaccinate the first trial participant the same day we received ethics approval. Time to enroll and randomize our target sample size of 380 patients was 22 days. CONCLUSION Taking the best of each system, we were able to flag eligible patients, transfer patient information automatically, randomize and enroll the patients in an easy workflow, decreasing the administrative burden usually associated with a trial of this size

Tumor Growth Enhances Cross-Presentation Leading to Limited T Cell Activation without Tolerance

Using a tumor model of spontaneously arising insulinomas expressing a defined tumor-associated antigen, we investigated whether tumor growth promotes cross-presentation and tolerance of tumor-specific T cells. We found that an advanced tumor burden enhanced cross-presentation of tumor-associated antigens to high avidity tumor-specific T cells, inducing T cell proliferation and limited effector function in vivo. However, contrary to other models, tumor-specific T cells were not tolerized despite a high tumor burden. In fact, in tumor-bearing mice, persistence and responsiveness of adoptively transferred tumor-specific T cells were enhanced. Accordingly, a potent T cell–mediated antitumor response could be elicited by intravenous administration of tumor-derived peptide and agonistic anti-CD40 antibody or viral immunization and reimmunization. Thus, in this model, tumor growth promotes activation of high avidity tumor-specific T cells instead of tolerance. Therefore, the host remains responsive to T cell immunotherapy

Somatic therapy of a mouse SMA model with a U7 snRNA gene correcting SMN2 splicing

Spinal Muscular Atrophy (SMA) is due to the loss of SMN1 gene function. The duplicate gene SMN2 produces some, but not enough, SMN protein because most transcripts lack exon 7. Thus, promoting the inclusion of this exon is a therapeutic option. We show that a somatic gene therapy using the gene for a modified U7 RNA which stimulates this splicing has a profound and persistent therapeutic effect on the phenotype of a severe SMA mouse model. To this end, the U7 gene and vector and the production of pure, highly concentrated self-complementary (sc) AAV9 vector particles were optimized. Introduction of the functional vector into motoneurons of newborn SMA mice by intracerebroventricular injection led to a highly significant, dose-dependent increase in life span and improvement of muscle functions. Besides the central nervous system, the therapeutic U7 RNA is expressed in the heart and liver which may additionally contribute to the observed therapeutic efficacy. This approach provides an additional therapeutic option for SMA and could also be adapted to treat other diseases of the central nervous systems with regulatory small RNA genes

Variability of Optical properties in perialpine lakes

The aim of the study is to describe how Inherent Optical Properties vary in perialpine lakes and to understand the impact of their vertical variability on the remote sensing signal