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Identifying colorectal cancer patient populations responsive to chemotherapy or chemoradiation therapy before surgery remains a challenge. Recently validated mouse protocols for organoid irradiation employ the single hit multi-target (SHMT) algorithm, which yields a single value, the D0, as a measure of inherent tissue radiosensitivity. Here, we translate these protocols to human tissue to evaluate radioresponsiveness of patient-derived organoids (PDO) generated from normal human intestines and rectal tumors of patients undergoing neoadjuvant therapy. While PDOs from adenomas with a logarithmically expanded Lgr5+ intestinal stem cell population retain the radioresistant phenotype of normal colorectal PDOs, malignant transformation yields PDOs from a large patient subpopulation displaying marked radiosensitivity due to reduced homologous recombination–mediated DNA repair. A proof-of-principle pilot clinical trial demonstrated that rectal cancer patient responses to neoadjuvant chemoradiation, including complete response, correlate closely with their PDO D0 values. Overall, upon transformation to colorectal adenocarcinoma, broad radiation sensitivity occurs in a large subset of patients that can be identified using SHMT analysis of PDO radiation responses.Significance:Analysis of inherent tissue radiosensitivity of patient-derived organoids may provide a readout predictive of neoadjuvant therapy response to radiation in rectal cancer, potentially allowing pretreatment stratification of patients likely to benefit from this approach.

Tissue survival responses to ionizing radiation are nonlinear with dose, rather yielding tissue-specific descending curves that impede straightforward analysis of biologic effects. Apoptotic cell death often occurs at low doses, while at clinically relevant intermediate doses, double-strand break misrepair yields mitotic death that determines outcome. As researchers frequently use a single low dose for experimentation, such strategies may inaccurately depict inherent tissue responses. Cutting edge radiobiology has adopted full dose survival profiling and devised mathematical algorithms to fit curves to observed data to generate highly reproducible numerical data that accurately define clinically relevant inherent radiosensitivities. Here, we established a protocol for irradiating organoids that delivers radiation profiles simulating the organ of origin. This technique yielded highly similar dose–survival curves of small and large intestinal crypts in vivo and their cognate organoids analyzed by the single-hit multi-target (SHMT) algorithm, outcomes reflecting the inherent radiation profile of their respective Lgr5+ stem cell populations. As this technological advance is quantitative, it will be useful for accurate evaluation of intestinal (patho)physiology and drug screening.Significance:These findings establish standards for irradiating organoids that deliver radiation profiles that phenocopy the organ of origin.See related commentary by Muschel et al., p. 927

Supplementary Data from Colorectal Cancer Develops Inherent Radiosensitivity That Can Be Predicted Using Patient-Derived Organoids

Supplementary Materials. Table S1. List of all reagents used in this study; Figure S1. LI organoids regrow after radiation; Figure S2. Sorting strategy for intestinal stem cells from the small and large intestines; Figure S3. Flow cytometry analysis of GFP+ cells in organoids versus colonies; Figure S4. Day 1 post-plating colonic stem cell colonies display defective DNA repair.

Persistent activation of the Hedgehog (HH)/GLI signaling pathway has been implicated in the development of a number of human cancers. The GLI zinc finger transcription factors act at the end of the HH signaling cascade to control gene expression, and recent studies have shown that the activity of GLI proteins can be additionally modified by integration of distinct signals, such as the MEK/extracellular signal-regulated kinase (ERK) and phosphinositide-3 kinase (PI3K)/AKT pathway. However, little is known about the identity of the upstream activators of these HH/GLI interacting signaling pathways in cancer. Here, we provide evidence that integration of the HH/GLI and epidermal growth factor receptor (EGFR) pathway synergistically induces oncogenic transformation, which depends on EGFR-mediated activation of the RAS/RAF/MEK/ERK but not of the PI3K/AKT pathway. EGFR/MEK/ERK signaling induces JUN/activator protein 1 activation, which is essential for oncogenic transformation, in combination with the GLI activator forms GLI1 and GLI2. Furthermore, pharmacologic inhibition of EGFR and HH/GLI efficiently reduces growth of basal cell carcinoma (BCC) cell lines derived from mice with activated HH/GLI signaling. The results identify the synergistic integration of GLI activator function and EGFR signaling as a critical step in oncogenic transformation and provide a molecular basis for therapeutic opportunities relying on combined inhibition of the HH/GLI and EGFR/MEK/ERK/JUN pathway in BCC. [Cancer Res 2009;69(4):1284–92]

Supplementary Figures 1-9, Tables 1-2 from Epidermal Growth Factor Receptor Signaling Synergizes with Hedgehog/GLI in Oncogenic Transformation via Activation of the MEK/ERK/JUN Pathway

Supplementary Methods, Tables 1-2 and Figures from Tumor Stroma–Derived Wnt5a Induces Differentiation of Basal Cell Carcinoma of Ptch-Mutant Mice via CaMKII

Supplementary Figures 1-9. Figure S1: Auto-phosphorylation of EGFR-A289V and EGFR-G598V can be inhibited by EGFR TKIs. Figure S2: EGFR* expression is strongly induced in hGFAP-tTA tetO-EGFR* off-Dox mouse brains. Figure S3: Gfap and Nestin protein expression are overlapped in a subpopulation of NPCs. Figure S4: Representative H&E images of a Grade IV malignant glioma. Figure S5: Malignant glioma cells do not express terminally differentiated CNS cell lineage markers. Figure S6: iEIP glioma subcutaneous transplants are sensitive to genetic suppression of EGFR* induction but not to EGFR TKI. Figure S7: iEIP gliomas are sensitive to acute EGFR* ablation. Figure S8: Ingenuity pathway analysis comparing gene expression profiling of untreated control tumors with Dox-treated relapsed tumors. Figure S9: Combined treatment of Dox and Bez-235 inhibits EGFR* induction and Akt activation.

Supplementary Figure Legends 1-9 from Epidermal Growth Factor Receptor Signaling Synergizes with Hedgehog/GLI in Oncogenic Transformation via Activation of the MEK/ERK/JUN Pathway

Identifying colorectal cancer patient populations responsive to chemotherapy or chemoradiation therapy before surgery remains a challenge. Recently validated mouse protocols for organoid irradiation employ the single hit multi-target (SHMT) algorithm, which yields a single value, the D0, as a measure of inherent tissue radiosensitivity. Here, we translate these protocols to human tissue to evaluate radioresponsiveness of patient-derived organoids (PDO) generated from normal human intestines and rectal tumors of patients undergoing neoadjuvant therapy. While PDOs from adenomas with a logarithmically expanded Lgr5+ intestinal stem cell population retain the radioresistant phenotype of normal colorectal PDOs, malignant transformation yields PDOs from a large patient subpopulation displaying marked radiosensitivity due to reduced homologous recombination–mediated DNA repair. A proof-of-principle pilot clinical trial demonstrated that rectal cancer patient responses to neoadjuvant chemoradiation, including complete response, correlate closely with their PDO D0 values. Overall, upon transformation to colorectal adenocarcinoma, broad radiation sensitivity occurs in a large subset of patients that can be identified using SHMT analysis of PDO radiation responses. Significance: Analysis of inherent tissue radiosensitivity of patient-derived organoids may provide a readout predictive of neoadjuvant therapy response to radiation in rectal cancer, potentially allowing pretreatment stratification of patients likely to benefit from this approach.

KurzfassungSedimentäre Strukturen können die Fließ- und Stofftransportpfade in feinkörnigen Talfüllungen stark beeinflussen. Diese Strukturen müssen gezielt auf ihre Ausdehnung und Eigenschaften untersucht werden, um Verweilzeiten, Fließpfade und das Abbaupotenzial eingetragener Schadstoffe zu bestimmen. In der quartären Talfüllung der Ammeraue bei Tübingen wurden beispielhaft Torflagen und eine Kiesrinne untersucht, um ihre Einflüsse auf die regionale Hydrogeologie und Hydrochemie zu bewerten. Dafür wurden geophysikalische und hydrogeologische Erkundungsmethoden ausgewählt und kombiniert. Mit geoelektrischen Oberflächenmessungen konnte die Ausdehnung der betrachteten Strukturen erkundet werden. Unterschiedliche Direct-Push-Sondierungen, darunter eine In-situ-Bestimmung der Sedimentfarbe, und bohrlochgeophysikalische Messungen erfassten ihre Geometrie und interne Heterogenität. Die hydraulischen und biogeochemischen Eigenschaften der Sedimente und des Grundwassers wurden anschließend durch gezielte Probennahmen und hydraulische Tests an repräsentativen Ansatzpunkten bestimmt. Die dargestellte Methodenkombination zur Abgrenzung relevanter Teilgebiete mit anschließender hochauflösender Untersuchung lässt sich auch auf die Untersuchung großflächiger Täler übertragen.

The Upper Neckar and Ammer River valleys in southwestern Germany correspond to the southwestern limit of the overall distribution of the oldest Linear Bandkeramik (LBK) culture. More than 200 Neolithic sites are known from this region, with one of the oldest sites located in the vicinity of the village Ammerbuch-Pfaffingen, 10 km west of Tubingen, Germany. The archaeological record suggests that settlement activities occurred here between approximately 6300 and 6030 BP (modelled ca. 5290-4900 calBC). Despite the various on-site activities, little is known about the environment and its resources that were available prior to and after the LBK arrival. We here present the first results of a palynological study of a 2.4 m section from two parallel, overlapping 16 m (in total) sediment cores. The cores were retrieved in 2018 from a palaeo-wetland (Ammer palaeo-mire) only 0.7 km distant from the LBK settlement “Lusse” and 2.5 km from the LBK settlement “Unteres Feld”. Pollen, spores, charcoal and plant macro-remains indicate three major periods of vegetational development between 10,650 and 7870 calBP. Accordingly, between 10,650 and 10,150 calBP, deciduous oak forests with strong participation of hazel (Corylus avellana) and open vegetation dominated by Artemisia, Chenopodiaceae and diverse species of the Asteraceae family were spread around a shallow palaeo-wetland of ~3 km2. From 10,150 to 8400 calBP, vegetation around the palaeo-wetland turns into a mixed oak forest with an even more prominent presence of hazel. From 8400 to 7870 calBP, a noticeable dominance of mixed oak forests is established in the surroundings and the palaeo-lake turns into a river floodplain. It is highly probable that, at the time of the arrival of the LBK, diverse natural plant resources were available from a mixture of trees, herbs and wetland taxa. The bioarchaeological evidence from the following LBK are based on the analysis of seeds/fruits and wood charcoal from ‘Lusse’ and ‘Unteres Feld’ and complements the information on land use for the period after the pollen record stops. The results suggest that the alluvial wetland area continued to provide food resources, together with the mixed oak forests which were also targeted by the LBK population. Apart from cultivation, LBK land-use caused an increase in the light-demanding forest component. The current study integrates the usually rarely available palaeo-ecological records from near LBK sites with on-site bioarchaeological evidence and thus delivers valuable insights on the environment at the beginning of farming in Central Europe.

Tissue survival responses to ionizing radiation are nonlinear with dose, rather yielding tissue-specific descending curves that impede straightforward analysis of biologic effects. Apoptotic cell death often occurs at low doses, while at clinically relevant intermediate doses, double-strand break misrepair yields mitotic death that determines outcome. As researchers frequently use a single low dose for experimentation, such strategies may inaccurately depict inherent tissue responses. Cutting edge radiobiology has adopted full dose survival profiling and devised mathematical algorithms to fit curves to observed data to generate highly reproducible numerical data that accurately define clinically relevant inherent radiosensitivities. Here, we established a protocol for irradiating organoids that delivers radiation profiles simulating the organ of origin. This technique yielded highly similar dose–survival curves of small and large intestinal crypts in vivo and their cognate organoids analyzed by the single-hit multi-target (SHMT) algorithm, outcomes reflecting the inherent radiation profile of their respective Lgr5+ stem cell populations. As this technological advance is quantitative, it will be useful for accurate evaluation of intestinal (patho)physiology and drug screening. Significance: These findings establish standards for irradiating organoids that deliver radiation profiles that phenocopy the organ of origin. See related commentary by Muschel et al., p. 927

Functional studies of primary cancer have been limited to animal models for a long time making it difficult to study aspects specific to human cancer biology. The development of organoid technology enabled us to culture human healthy and tumor cells as three-dimensional self-organizing structures in vitro for a prolonged time. Organoid cultures conserve the heterogeneity of the originating epithelium regarding cell types and tumor clonality. Therefore, organoids are considered an invaluable tool to study and genetically dissect various aspects of human cancer biology. In this review, we describe the applications, advantages, and limitations of organoids as human cancer models with the main emphasis on colorectal cancer.

Recent data establish a logarithmic expansion of leucine rich repeat containing G protein coupled receptor 5-positive (Lgr5+) colonic epithelial stem cells (CESCs) in human colorectal cancer (CRC). Complementary studies using the murine 2-stage azoxymethane-dextran sulfate sodium (AOM-DSS) colitis-associated tumor model indicate early acquisition of Wnt pathway mutations drives CESC expansion during adenoma progression. Here, subdivision of the AOM-DSS model into in vivo and in vitro stages revealed DSS induced physical separation of CESCs from stem cell niche cells and basal lamina, a source of Wnt signals, within hours, disabling the stem cell program. While AOM delivery in vivo under non-adenoma-forming conditions yielded phenotypically normal mucosa and organoids derived thereof, niche injury ex vivo by progressive DSS dose escalation facilitated outgrowth of Wnt-independent dysplastic organoids. These organoids contained 10-fold increased Lgr5+ CESCs with gain-of-function Wnt mutations orthologous to human CRC driver mutations. We posit CRC originates by niche injury-induced outgrowth of normally suppressed mutated stem cells, consistent with models of adaptive oncogenesis.

Tumor cure with conventional fractionated radiotherapy is 65%, dependent on tumor cell-autonomous gradual buildup of DNA double-strand break (DSB) misrepair. Here we report that single-dose radiotherapy (SDRT), a disruptive technique that ablates more than 90% of human cancers, operates a distinct dual-target mechanism, linking acid sphingomyelinase-mediated (ASMase-mediated) microvascular perfusion defects to DNA unrepair in tumor cells to confer tumor cell lethality. ASMase-mediated microcirculatory vasoconstriction after SDRT conferred an ischemic stress response within parenchymal tumor cells, with ROS triggering the evolutionarily conserved SUMO stress response, specifically depleting chromatin-associated free SUMO3. Whereas SUMO3, but not SUMO2, was indispensable for homology-directed repair (HDR) of DSBs, HDR loss of function after SDRT yielded DSB unrepair, chromosomal aberrations, and tumor clonogen demise. Vasoconstriction blockade with the endothelin-1 inhibitor BQ-123, or ROS scavenging after SDRT using peroxiredoxin-6 overexpression or the SOD mimetic tempol, prevented chromatin SUMO3 depletion, HDR loss of function, and SDRT tumor ablation. We also provide evidence of mouse-to-human translation of this biology in a randomized clinical trial, showing that 24 Gy SDRT, but not 3×9 Gy fractionation, coupled early tumor ischemia/reperfusion to human cancer ablation. The SDRT biology provides opportunities for mechanism-based selective tumor radiosensitization via accessing of SDRT/ASMase signaling, as current studies indicate that this pathway is tractable to pharmacologic intervention.

In soils and sediments, large amounts of total organic carbon (TOC) mark reducing conditions. As dark sediment colors are good predictors for high‐TOC zones, they indicate hot spots of biogeochemical turnover and microbial activity. Traditionally, obtaining the sediment color or TOC at depth requires costly core sampling, resulting in poor horizontal resolution and related uncertainty caused by interpolation. We suggest using a direct‐push tool for optical screening of the sediment color to acquire multiple high‐resolution vertical color profiles and demonstrate its applicability to a biogeochemical transition zone in floodplain sediments, dominated by tufa. We use Gaussian mixture models for a cluster analysis of 35 color logs in the International Commission on Illumination (CIE) L*a*b* color space to identify three colorfacies that differ in lithology and TOC content: a dark colorfacies that agrees well with peat layers, a gray colorfacies associated with clay, and a creamy‐brown facies made of autochthonous carbonate precipitates. We test different approaches either to infer the TOC content from color metrics, namely, the lightness and chroma, across all facies, or to identify TOC ranges for each colorfacies. Given the high variability in TOC due to organic carbon specks in the tufa, the latter approach appears more realistic. In our application we map the 3‐D distribution of organic matter in a floodplain in distinct facies over 20,000 m2 down to 12 m depth. While we relate the sediment color only to the TOC content, direct‐push color logging may also be used for in situ mapping of other biogeochemically relevant properties, such as the ferric‐iron content or sedimentary structure., Plain Language Summary: Geologists can say a lot about soils and loose materials in the ground by looking at their color. Dark materials normally contain dead plants, called organic carbon, which are food for bacteria and cause chemical reactions. To get the color of the soil, geologists normally need soil samples, but getting them from depth takes time and money. We test a method of pushing a camera into the ground and recording the color therein. From the recorded color we can say which type of geological material is at which depth and how much organic carbon is there without taking samples everywhere. This can be done very quickly, so that we can do it over a large area and down to the depth of the loose material in the ground, where most of the groundwater flows., Key Points: Direct‐push methods allow vertical in situ profiling of sediment color at high resolution. Cluster analysis reproducibly delineates colorfacies indicative of sedimentary units. Site‐specific relationships between color and TOC in sediments permit identifying biogeochemical hot spots., German Research Foundation (Deutsche Forschungsgemeinschaft, DFG)

The Upper Neckar and Ammer river valleys in southwestern Germany correspond to the southwestern limit of the overall distribution of the oldest Linear Bandkeramik (LBK) culture. More than 200 Neolithic sites are known from this region with one of the oldest sites located in the vicinity of the village Ammerbuch-Pfäffingen, around 10 km west of Tübingen, Germany. The archaeological record suggests that settlement activities occurred here between 7260 and 7110 cal BP (or 5310-5160 cal BC). Despite the various activities at the settling site itself, little is known about the environmental impact of the first settlers on the area, ranging from the introduction of farming and animal husbandry with impacts to the forests due to pasture and collection of wood, as well as possible control of water bodies. We here present the first results of a palynological study of two parallel, overlapping 8 m long sediment cores that were retrieved in 2018 from a shallow paleo-lake only a few hundred meters distant from the excavation site. The composite core allows environmental reconstruction of the area between 11540 and 7000 cal BP, based on six radiocarbon dates. Pollen analysis indicates mixed oak forests and an increase of light-demanding vegetation (i.e. Quercus, Corylus, Betula). Current analyses on micro- and macro-charcoal are going to reveal the natural or anthropogenic induced causes of paleo-fire events and Non-Pollen-Palynomorphs (NPP), including dung spores, unravel the possible presence of herbivores (including domestic ones) in the area. The results of the current study and its integration into the bioarchaeological record are relevant even beyond the region providing the usually rarely available paleoecological records from close proximity of an LBK site and thus deliver valuable insights on the environmental settings at the beginning of farming in central Europe.

Recent evidence suggests that mammary cells expressing R-spondin receptor and Wnt pathway regulator Lgr5, regarded as a stem cell marker in multiple tissues, might represent mammary stem cells (MaSCs). Whether L gr5 marks a multipotent subpopulation of Lin-CD24low/medCD49fhigh MaSCs remains controversial. To some extent the differing results reflect different assays used to assess properties of stemness, including lineage tracing in vivo, mammosphere culture, and mammary fat pad transplantation assays. To address this issue directly, we isolated Lgr5+ cells from mammary glands of Lgr5-lacZ mice and established organoids based on principles adapted from studies of Wnt-driven Lgr5+ cell populations in other organs. Mammary organoids were grown from single Lgr5+ mammary cells in Matrigel, the substratum of choice for intestinal organoids, and in a growth factor cocktail containing EGF, Wnt3a and R-spondin, designed to optimally activate the endogenous Wnt signaling program of stem cells. Colonies derived from single Lgr5+ cells manifest at least four distinct cell populations: Lgr5+ and Lgr5− basal cells and c-Kit+ and c-Kit− luminal cells that spontaneously organize into a ductal structure with basal cells around the periphery and luminal cells lining an interior cavity, reminiscent of normal mammary duct structure. Lgr5+ cell-derived organoids were sustainable during prolonged passaging. In contrast, although Lgr5− cells expand into primary colonies, colony-forming efficiency immediately dissipated upon passaging. Furthermore, reproductive hormones induce epithelial cell proliferation resulting in marked increases in lumen diameter accompanied by squamous transdifferentiation. We propose this estrogen-responsive, self-organizing duct-like structure derived from single murine Lgr5+ mammary cells represents a “mini-breast” organoid.

Efforts to identify and target glioblastoma (GBM) drivers have primarily focused on receptor tyrosine kinases (RTKs). Clinical benefits, however, have been elusive. Here, we identify a SRY-related box 2 (SOX2) transcriptional regulatory network that is independent of upstream RTKs and is capable of driving glioma initiating cells. We identified oligodendrocyte lineage transcription factor 2 (OLIG2) and zinc finger E-box binding homeobox 1 (ZEB1) as potential SOX2 targets, which are frequently co-expressed irrespective of driver mutations. In murine glioma models, we show that different combinations of tumor suppressor and oncogene mutations can activate Sox2, Olig2, and Zeb1 expression. We demonstrate that ectopic co-expression of the three transcription factors can transform tumor suppressor deficient astrocytes into glioma initiating cells in the absence of an upstream RTK oncogene. Finally, we demonstrate that the transcriptional inhibitor mithramycin downregulates SOX2 and its target genes, resulting in markedly reduced proliferation of GBM cells in vivo.

In most hydrogeological, geotechnical, archaeological, and other geoscientific applications, we need to understand the lateral extent and connectivity of system-relevant subsurface features. Towards this end, direct-current electric resistivity tomography (ERT) with several 2-D profiles or 3-D grids provides a powerful tool for non-invasive resolution of electrical resistivity anomalies. On the downside, many hours of fieldwork to set up and break down long electrode profiles limit this method to study areas of few thousand square meters, as the workload multiplies with the number of profiles. In many projects, however, determining the extent and connectivity of subsurface anomalies and therefore their potential relevance to the system, may only require the target to be spatially traced instead of fully resolved. We therefore propose geoelectric mapping with a target-specific fixed electrode spacing as an efficient way to trace a resolved resistivity anomaly away from an initial ERT profile, which should be particularly valuable for large study areas. The target-specific electrode spacing is hereby determined by evaluating the effects of the targeted anomaly in the raw data of the preliminary ERT profile. We therefore introduce an anomaly effect applicable to measurements in environments with spatial trends in resistivity distribution. In synthetic simulations, we demonstrate that our approach can efficiently delineate lateral boundaries of resistivity anomalies in ERT data space and we visualize this in pseudosections of anomaly effects. We then apply this method to tracing a gravel-filled paleo-channel in the 8 km2 Ammer floodplain near Tubingen, Germany and determine a suitable electrode spacing for a subsequent mapping campaign from the ranges of anomaly effects. We traced the paleo-channel over several hundreds of meters away from an initial 550 m long ERT profile within 19 h, the same time needed to set up, measure, and dismantle the single initial ERT profile. The evaluation of anomaly effects proves to be an efficient tool to detect resistivity anomalies in geoelectric data and determine suitable electrode spacings for large-scale mapping campaigns. Once identified, anomalies and project-relevant subareas can be the target of more detailed investigations.

We investigate yttrium iron garnet (YIG)/cobalt (Co) heterostructures using broadband ferromagnetic resonance (FMR). We observe an efficient excitation of perpendicular standing spin waves (PSSWs) in the YIG layer when the resonance frequencies of the YIG PSSWs and the Co FMR line coincide. Avoided crossings of YIG PSSWs and the Co FMR line are found and modeled using mutual spin pumping and exchange torques. The excitation of PSSWs is suppressed by a thin aluminum oxide (AlOx) interlayer but persists with a copper (Cu) interlayer, in agreement with the proposed model.

We experimentally study the magnon-photon coupling in a system consitsing of the compensating ferrimagnet gadolinium iron garnet (GdIG) and a three-dimensional microwave cavity. The temperature is varied in order to tune the GdIG magnetization and to observe the transition from the weak coupling regime to the strong coupling regime. By measuring and modelling the complex reflection parameter of the system the effective coupling rate g eff and the magnetization M eff of the sample are extracted. Comparing g eff with the magnon and the cavity decay rate we conclude that the strong coupling regime is easily accessible using GdIG. We show that the effective coupling strength follows the predicted square root dependence on the magnetization.

We investigate the temperature-dependent microwave absorption spectrum of an yttrium iron garnet sphere as a function of temperature (5 to $300\phantom{\rule{4pt}{0ex}}\mathrm{K}$) and frequency (3 to $43.5\phantom{\rule{4pt}{0ex}}\mathrm{GHz}$). At temperatures above $100\phantom{\rule{4pt}{0ex}}\mathrm{K}$, the magnetic resonance linewidth increases linearly with temperature and shows a Gilbert-like linear frequency dependence. At lower temperatures, the temperature dependence of the resonance linewidth at constant external magnetic fields exhibits a characteristic peak which coincides with a non-Gilbert-like frequency dependence. The complete temperature and frequency evolution of the linewidth can be modeled by the phenomenology of slowly relaxing rare-earth impurities and either the Kasuya-LeCraw mechanism or the scattering with optical magnons. Furthermore, we extract the temperature dependence of the saturation magnetization, the magnetic anisotropy, and the $g$ factor.

We studied the spin injection in a NiFe(Py)/Si system using broadband ferromagnetic resonance spectroscopy. The Gilbert damping parameter of the Py layer on top of the Si channel was determined as a function of the Si doping concentration and Py layer thickness. For fixed Py thickness we observed an increase of the Gilbert damping parameter with decreasing resistivity of the Si channel. For a fixed Si doping concentration we measured an increasing Gilbert damping parameter for decreasing Py layer thickness. No increase of the Gilbert damping parameter was found Py/Si samples with an insulating interlayer. We attribute our observations to an enhanced spin injection into the low-resistivity Si by spin pumping., Comment: 15 pages, 3 Figures, 2 Tables (to appear Applied Physics Letters)

A d-orbital electron has an anisotropic electron orbital and is a source of magnetism. The realization of a 2-dimensional electron gas (2DEG) embedded at a LaAlO3/SrTiO3 interface surprised researchers in materials and physical sciences because the 2DEG consists of 3d-electrons of Ti with extraordinarily large carrier mobility, even in the insulating oxide heterostructure. To date, a wide variety of physical phenomena, such as ferromagnetism and the quantum Hall effect, have been discovered in this 2DEG systems, demonstrating the ability of the d-electron 2DEG systems to provide a material platform for the study of interesting physics. However, because of both ferromagnetism and the Rashba field, long-range spin transport and the exploitation of spintronics functions have been believed difficult to implement in the d-electron 2DEG systems. Here, we report the experimental demonstration of room-temperature spin transport in the d-electron-based 2DEG at a LaAlO3/SrTiO3 interface, where the spin relaxation length is ca. exceeding 200 nm. Our finding, which counters the conventional understandings to d-electron 2DEGs, opens a new field of d-electron spintronics. Furthermore, this work highlights a novel spin function in the conductive oxide system., 30 pages, 5 Figures, 1 Table, including Supplemental Information

A d-orbital electron has an anisotropic electron orbital and is a source of magnetism. The realization of a two-dimensional electron gas (2DEG) embedded at a LaAlO

Boolean logic is the foundation of modern digital information processing. Recently, there has been a growing interest in phenomena based on pure spin currents, which allow to move from charge to spin based logic gates. We study a proof-of-principle logic device based on the ferrimagnetic insulator Yttrium Iron Garnet (YIG), with Pt strips acting as injectors and detectors for nonequilibrium magnons. We experimentally observe incoherent superposition of magnons generated by different injectors. This allows to implement a fully functional majority gate, enabling multiple logic operations (AND and OR) in one and the same device. Clocking frequencies of the order of several GHz and straightforward down-scaling make our device promising for applications.

Microfocused Brillouin light scattering (BLS) and microwave absorption (MA) are used to study magnon-photon coupling in a system consisting of a split-ring microwave resonator and a yttrium iron garnet (YIG) film. The split-ring resonantor is defined by optical lithography and loaded with a 1 $\mu$m-thick YIG film grown by liquid phase epitaxy. BLS and MA spectra of the hybrid system are simultaneously recorded as a function of the applied magnetic field magnitude and microwave excitation frequency. Strong coupling of the magnon and photon modes is found with a coupling strength of $g_\text{eff}/2 \pi = 63$ MHz. The combined BLS and MA data allows to study the continuous transition of the hybridized modes from a purely magnonic to a purely photonic mode by varying the applied magnetic field and microwave frequency. Furthermore, the BLS data represent an up-conversion of the microwave frequency coupling to optical frequencies., Comment: 5 pages, 3 figures

Inhibition of Hedgehog (HH)/GLI signalling in cancer is a promising therapeutic approach. Interactions between HH/GLI and other oncogenic pathways affect the strength and tumourigenicity of HH/GLI. Cooperation of HH/GLI with epidermal growth factor receptor (EGFR) signalling promotes transformation and cancer cell proliferation in vitro. However, the in vivo relevance of HH-EGFR signal integration and the critical downstream mediators are largely undefined. In this report we show that genetic and pharmacologic inhibition of EGFR signalling reduces tumour growth in mouse models of HH/GLI driven basal cell carcinoma (BCC). We describe HH-EGFR cooperation response genes including SOX2, SOX9, JUN, CXCR4 and FGF19 that are synergistically activated by HH-EGFR signal integration and required for in vivo growth of BCC cells and tumour-initiating pancreatic cancer cells. The data validate EGFR signalling as drug target in HH/GLI driven cancers and shed light on the molecular processes controlled by HH-EGFR signal cooperation, providing new therapeutic strategies based on combined targeting of HH-EGFR signalling and selected downstream target genes.

Basal cell carcinoma (BCC) is the most common skin tumor in humans. Although BCCs rarely metastasize, they can cause significant morbidity due to local aggressiveness. Approximately 20% of BCCs show signs of spontaneous regression. The understanding of molecular events mediating spontaneous regression has the potential to reduce morbidity of BCC and, potentially, other tumors, if translated into tumor therapies. We show that BCCs induced in conditional Ptchflox/floxERT2+/− knockout mice regress with time and show a more differentiated phenotype. Differentiation is accompanied by Wnt5a expression in the tumor stroma, which is first detectable at the fully developed tumor stage. Coculture experiments revealed that Wnt5a is upregulated in tumor-adjacent macrophages by soluble signals derived from BCC cells. In turn, Wnt5a induces the expression of the differentiation marker K10 in tumor cells, which is mediated by Wnt/Ca2+ signaling in a CaMKII-dependent manner. These data support a role of stromal Wnt5a in BCC differentiation and regression, which may have important implications for development of new treatment strategies for this tumor. Taken together, our results establish BCC as an easily accessible model of tumor regression. The regression of BCC despite sustained Hedgehog signaling activity seems to be mediated by tumor-stromal interactions via Wnt5a signaling. Cancer Res; 70(7); 2739–48

Hedgehog (HH) signaling in the epidermis is primarily mediated by the zinc finger transcription factors GLI1 and GLI2. Exquisite regulation of HH/GLI signaling is crucial for proper specification of the epidermal lineage and development of its derivatives, whereas dysregulation of HH/GLI signaling disrupts tissue homeostasis and causes basal cell carcinoma (BCC). Similarly, bone morphogenetic proteins (BMPs) and activins have been described as key signaling factors in the complex regulation of epidermal fate decisions, although their precise interplay with HH/GLI is largely elusive. Here we show that, in human epidermal cells, expression of the activin/BMP antagonist follistatin (FST) is predominantly up-regulated by the HH effector GLI2. Consistently, we found strong FST expression in the outer root sheath of human hair follicles and BCC. Detailed promoter analysis showed that two sequences with homology to the GLI consensus binding site are required for GLI2-mediated activation. Interestingly, activation of the FST promoter is highly GLI2-specific, because neither GLI1 nor GLI3 can significantly increase FST transcription. GLI2 specificity requires the presence of a 518-bp fragment in the proximal FST promoter region. On the protein level, sequences C-terminal to the zinc finger are responsible for GLI2-specific activation of FST transcription, pointing to the existence of GLI-interacting cofactors that modulate GLI target specificity. Our results reveal a key role of GLI2 in activation of the activin/BMP antagonist FST in response to HH signaling and provide new evidence for a regulatory interaction between HH and activin/BMP signaling in hair follicle development and BCC.

The magnetostatic mode (MSM) spectrum of a 300$\mu$m diameter single crystalline sphere of yttrium iron garnet is investigated using broadband ferromagnetic resonance (FMR). The individual MSMs are identified via their characteristic dispersion relations and the corresponding mode number tuples $(nmr)$ are assigned. Taking FMR data over a broad frequency and magnetic field range allows to analyze both the Gilbert damping parameter~$\alpha$ and the inhomogeneous line broadening contribution to the total linewidth of the MSMs separately. The linewidth analysis shows that all MSMs share the same Gilbert damping parameter $\alpha=2.7(5) \times 10^{-5}$ irrespective of their mode index. In contrast, the inhomogeneous line broadening shows a pronounced mode dependence. This observation is modeled in terms of two-magnon scattering processes of the MSMs into the spin-wave manifold, mediated by surface and volume defects., Comment: 5 pages, 3 figures

Epidermal growth factor receptor (EGFR) is highly amplified, mutated, and overexpressed in human malignant gliomas. Despite its prevalence and growth-promoting functions, therapeutic strategies to inhibit EGFR kinase activity have not been translated into profound beneficial effects in glioma clinical trials. To determine the roles of oncogenic EGFR signaling in gliomagenesis and tumor maintenance, we generated a novel glioma mouse model driven by inducible expression of a mutant EGFR (EGFR*). Using combined genetic and pharmacologic interventions, we revealed that EGFR*-driven gliomas were insensitive to EGFR tyrosine kinase inhibitors, although they could efficiently inhibit EGFR* autophosphorylation in vitro and in vivo. This is in contrast with the genetic suppression of EGFR* induction that led to significant tumor regression and prolonged animal survival. However, despite their initial response to genetic EGFR* extinction, all tumors would relapse and propagate independent of EGFR*. We further showed that EGFR*-independent tumor cells existed prior to treatment and were responsible for relapse following genetic EGFR* suppression. And, the addition of a PI3K/mTOR inhibitor could significantly delay relapse and prolong animal survival. Our findings shed mechanistic insight into EGFR drug resistance in glioma and provide a platform to test therapies targeting aberrant EGFR signaling in this setting. Cancer Res; 75(10); 2109–19. ©2015 AACR.

The design of a microstructured, fully functional spin-wave majority gate is presented and studied using micromagnetic simulations. This all-magnon logic gate consists of three-input waveguides, a spin-wave combiner and an output waveguide. In order to ensure the functionality of the device, the output waveguide is designed to perform spin-wave mode selection. We demonstrate that the gate evaluates the majority of the input signals coded into the spin-wave phase. Moreover, the all-magnon data processing device is used to perform logic AND-, OR-, NAND- and NOR- operations., Comment: 5 pages, 5 figures

Persistent activation of the Hedgehog (HH)/GLI signaling pathway has been implicated in the development of a number of human cancers. The GLI zinc finger transcription factors act at the end of the HH signaling cascade to control gene expression, and recent studies have shown that the activity of GLI proteins can be additionally modified by integration of distinct signals, such as the MEK/extracellular signal-regulated kinase (ERK) and phosphinositide-3 kinase (PI3K)/AKT pathway. However, little is known about the identity of the upstream activators of these HH/GLI interacting signaling pathways in cancer. Here, we provide evidence that integration of the HH/GLI and epidermal growth factor receptor (EGFR) pathway synergistically induces oncogenic transformation, which depends on EGFR-mediated activation of the RAS/RAF/MEK/ERK but not of the PI3K/AKT pathway. EGFR/MEK/ERK signaling induces JUN/activator protein 1 activation, which is essential for oncogenic transformation, in combination with the GLI activator forms GLI1 and GLI2. Furthermore, pharmacologic inhibition of EGFR and HH/GLI efficiently reduces growth of basal cell carcinoma (BCC) cell lines derived from mice with activated HH/GLI signaling. The results identify the synergistic integration of GLI activator function and EGFR signaling as a critical step in oncogenic transformation and provide a molecular basis for therapeutic opportunities relying on combined inhibition of the HH/GLI and EGFR/MEK/ERK/JUN pathway in BCC. [Cancer Res 2009;69(4):1284–92]

We propose the utilization of isotropic forward volume magneto-static spin waves in modern wave-based logic devices and suggest a concrete design for a spin-wave majority gate operating with these waves. We demonstrate by numerical simulations that the proposed out-of-plane magnetized majority gate overcomes the limitations of anisotropic in-plane magnetized majority gates due to the high spin-wave transmission through the gate, which enables a reduced energy consumption of these devices. Moreover, the functionality of the out-of-plane majority gate is increased due to the lack of parasitic generation of short-wavelength exchange spin waves., Comment: 4 pages, 4 figures

Hedgehog (HH)/GLI signaling plays a critical role in epidermal development and basal cell carcinoma. Here, we provide evidence that epidermal growth factor receptor (EGFR) signaling modulates the target gene expression profile of GLI transcription factors in epidermal cells. Using expression profiling and quantitative reverse transcriptase PCR, we identified a set of 19 genes whose transcription is synergistically induced by GLI1 and parallel EGF treatment. Promoter studies of a subset of GLI/EGF-regulated genes, including the genes encoding interleukin-1 antagonist IL1R2, Jagged 2, cyclin D1, S100A7, and S100A9, suggest convergence of EGFR and HH/GLI signaling at the level of promoters of selected direct GLI target genes. Inhibition of EGFR and MEK/ERK but not of phosphatidylinositol 3-kinase/AKT abrogated synergistic activation of GLI/EGF target genes, showing that EGFR can signal via RAF/MEK/ERK to cooperate with GLI proteins in selective target gene regulation. Coexpression of the GLI/EGF target IL1R2, EGFR, and activated ERK1/2 in human anagen hair follicles argues for a cooperative role of EGFR and HH/GLI signaling in specifying the fate of outer root sheath (ORS) cells. We also show that EGF treatment neutralizes GLI-mediated induction of epidermal stem cell marker expression and provide evidence that EGFR signaling is essential for GLI-induced cell cycle progression in epidermal cells. The results suggest that EGFR signaling modulates GLI target gene profiles which may play an important regulatory role in ORS specification, hair growth, and possibly cancer.

We present a study of the influence of the annealing temperature Ta on the magnetic properties of Co40Fe40B20 thin films. Using a vector network analyzer ferromagnetic resonance (VNA-FMR) setup and the magneto-optical Kerr effect, the dependence of the Gilbert damping parameter α, the exchange constant A, the saturation magnetization as well as the coercive field and the Kerr signal on Ta is reported. Additionally, the correlation with the crystalline properties of the films studied by X-ray diffractometry is discussed. We found that while the damping parameter α and the coercive field show sharp changes starting at a certain Ta value, the exchange constant A and the Kerr signal show a steady evolution. A differential modification of the film surface compared to the bulk is discussed as a possible reason. In any case, we found that the low damping values are preserved at the first onset of crystallization.

A characterization of the magnetic properties of amorphous Co40Fe40B20 thin films, developed for low damping applications in magnon spintronics, using vector network analyzer ferromagnetic resonance (VNA-FMR) and the magneto-optical Kerr effect is presented. Our films show a very weak uniaxial anisotropy and a low Gilbert damping parameter (α=0.0042). The saturation magnetization MS extracted from the FMR measurements is 1250 kA/m. The frequency dependence of the first perpendicular standing spin waves mode on the applied magnetic field is used to determine the exchange constant A for this alloy resulting in a value of 1.5×10−11 J/m. These values are discussed in comparison to literature values for different CoFeB compositions and other related alloys.