The optogenetic promise for oncology: Episode I

As light-based control of fundamental signaling pathways is becoming a reality, the field of optogenetics is rapidly moving beyond neuroscience. We have recently developed receptor tyrosine kinases that are activated by light and control cell proliferation, epithelial–mesenchymal transition, and angiogenic sprouting—cell behaviors central to cancer progression

A phytochrome sensory domain permits receptor activation by red light

Optogenetics and photopharmacology enable the spatio-temporal control of cell and animal behavior by light. Although red light offers deep-tissue penetration and minimal phototoxicity, very few red-light-sensitive optogenetic methods are currently available. We have now developed a red-light-induced homodimerization domain. We first showed that an optimized sensory domain of the cyanobacterial phytochrome 1 can be expressed robustly and without cytotoxicity in human cells. We then applied this domain to induce the dimerization of two receptor tyrosine kinases—the fibroblast growth factor receptor 1 and the neurotrophin receptor trkB. This new optogenetic method was then used to activate the MAPK/ERK pathway non-invasively in mammalian tissue and in multicolor cell-signaling experiments. The light-controlled dimerizer and red-light-activated receptor tyrosine kinases will prove useful to regulate a variety of cellular processes with light. Go deep with red: The sensory domain (S) of the cyanobacterial phytochrome 1 (CPH1) was repurposed to induce the homodimerization of proteins in living cells by red light. By using this domain, light-activated protein kinases were engineered that can be activated orthogonally from many fluorescent proteins and through mammalian tissue. Pr/Pfr=red-/far-red-absorbing state of CPH1

Light-assisted small-molecule screening against protein kinases

High-throughput live-cell screens are intricate elements of systems biology studies and drug discovery pipelines. Here, we demonstrate an optogenetics-assisted method that avoids the need for chemical activators and reporters, reduces the number of operational steps and increases information content in a cell-based small-molecule screen against human protein kinases, including an orphan receptor tyrosine kinase. This blueprint for all-optical screening can be adapted to many drug targets and cellular processes

Measurement and simulation of the muon-induced neutron yield in lead

A measurement is presented of the neutron production rate in lead by high energy cosmic-ray muons at a depth of 2850m water equivalent (w.e.) and a mean muon energy of 260GeV. The measurement exploits the delayed coincidences between muons and the radiative capture of induced neutrons in a highly segmented tonne scale plastic scintillator detector. Detailed Monte Carlo simulations reproduce well the measured capture times and multiplicities and, within the dynamic range of the instrumentation, the spectrum of energy deposits. By comparing measurements with simulations of neutron capture rates a neutron yield in lead of (5.78_-_0_._2_8^+^0^.^2^1) x10^-^3neutrons/muon/(g/cm^2) has been obtained. Absolute agreement between simulation and data is of order 25%. Consequences for deep underground rare event searches are discussed.Peer Reviewe

Grünlicht-induzierte Rezeptorinaktivierung durch Cobalamin-bindende Domänen

Optogenetik und Photopharmakologie ermöglichen präzise räumliche und zeitliche Kontrolle von Proteinwechselwirkung und -funktion in Zellen und Tieren. Optogenetische Methoden, die auf grünes Licht ansprechen und zum Trennen von Proteinkomplexen geeignet sind, sind nichtweitläufig verfügbar, würden jedoch mehrfarbige Experimente zur Beantwortung von biologischen Fragestellungen ermöglichen. Hier demonstrieren wir die Verwendung von Cobalamin(Vitamin B12)-bindenden Domänen von bakteriellen CarH-Transkriptionsfaktoren zur Grünlicht-induzierten Dissoziation von Rezeptoren. Fusioniert mit dem Fibroblasten-W achstumsfaktor-Rezeptor 1 führten diese im Dunkeln in kultivierten Zellen zu Signalaktivität durch Oligomerisierung, welche durch Beleuchten umgehend aufgehoben wurde. In Zebrafischembryonen, die einen derartigen Rezeptor exprimieren, ermöglichte grünes Licht die Kontrolle über abnormale Signalaktivität während der Embryonalentwicklung

The dorsoventral regulatory gene cassette spätzle/Toll/cactus controls the potent antifungal response in Drosophila adults

The cytokine-induced activation cascade of NF-kappaB in mammals and the activation of the morphogen dorsal in Drosophila embryos show striking structural and functional similarities (Toll/IL-1, Cactus/I-kappaB, and dorsal/NF-kappaB). Here we demonstrate that these parallels extend to the immune response of Drosophila. In particular, the intracellular components of the dorsoventral signaling pathway (except for dorsal) and the extracellular Toll ligand, spätzle, control expression of the antifungal peptide gene drosomycin in adults. We also show that mutations in the Toll signaling pathway dramatically reduce survival after fungal infection. Antibacterial genes are induced either by a distinct pathway involving the immune deficiency gene (imd) or by combined activation of both imd and dorsoventral pathways

WIMP-nucleon cross-section results from the second science run of ZEPLIN-III

We report experimental upper limits on WIMP-nucleon elastic scattering cross sections from the second science run of ZEPLIN-III at the Boulby Underground Laboratory. A raw fiducial exposure of 1,344 kg.days was accrued over 319 days of continuous operation between June 2010 and May 2011. A total of eight events was observed in the signal acceptance region in the nuclear recoil energy range 7-29 keV, which is compatible with background expectations. This allows the exclusion of the scalar cross-section above 4.8E-8 pb near 50 GeV/c^2 WIMP mass with 90% confidence. Combined with data from the first run, this result improves to 3.9E-8 pb. The corresponding WIMP-neutron spin-dependent cross-section limit is 8.0E-3 pb. The ZEPLIN programme reaches thus its conclusion at Boulby, having deployed and exploited successfully three liquid xenon experiments of increasing reach

Quenching Factor for Low Energy Nuclear Recoils in a Plastic Scintillator

Plastic scintillators are widely used in industry, medicine and scientific research, including nuclear and particle physics. Although one of their most common applications is in neutron detection, experimental data on their response to low-energy nuclear recoils are scarce. Here, the relative scintillation efficiency for neutron-induced nuclear recoils in a polystyrene-based plastic scintillator (UPS-923A) is presented, exploring recoil energies between 125 keV and 850 keV. Monte Carlo simulations, incorporating light collection efficiency and energy resolution effects, are used to generate neutron scattering spectra which are matched to observed distributions of scintillation signals to parameterise the energy-dependent quenching factor. At energies above 300 keV the dependence is reasonably described using the semi-empirical formulation of Birks and a kB factor of (0.014+/-0.002) g/MeVcm^2 has been determined. Below that energy the measured quenching factor falls more steeply than predicted by the Birks formalism.Comment: 8 pages, 9 figure