Charakterisierung von Rhabdomyosarkomen und Etablierung einer adoptiven T-Zelltherapie auf der Grundlage chimärer T-Zellen

Rhabdomyosarkome (RMS) sind die häufigsten Weichgewebetumoren bei Kindern und Jugendlichen. Obwohl die 5-Jahres-Überlebensrate bei lokalisierten RMS heute 70% erreicht, ist die Überlebensrate bei metastasierten RMS gering. Diesen Risikopatienten eröffnen sich neue Perspektiven durch immuntherapeutische Strategien. Die Entwicklung des adoptiven Transfers RMS-reaktiver chimärer T Zellen steht im Zentrum der vorliegenden Arbeit. In Voruntersuchungen wurden chimäre Antigenrezeptoren (CAR) mit Spezifität für ein RMS spezifisches Antigen, den fetalen Acetylcholinrezeptor (fAChR), generiert. Die retrovirale Expression dieses CAR auf der Oberfläche von Blutlymphozyten erzeugt chimäre T Zellen, die fAChR-exprimierende RMS-Zellen in vitro und in vivo erkennen. CAR der ersten Generation, die aus dem scFv-Fragment eines humanen fAChR spezifischen Autoantikörpers, einer IgG und einer CD3ζ Domäne bestehen, erzeugen in chimären T Zellen jedoch eine zu geringe zytolytische Aktivität, um ihren Einsatz in vivo zu rechtfertigen. Im Rahmen der vorliegenden Arbeit wurde daher der CAR der ersten Generation mit einer zusätzlichen CD28 Signaldomäne ausgestattet, um fehlende kostimulatorische Signale zu kompensieren und die Anergisierung chimärer Zellen zu verhindern. Diese Rezeptor-Modifikation steigerte die lytische Aktivität der chimären Zellen signifikant, blieb jedoch im Vergleich zu CEA spezifischen CAR bei Colon Karzinomzellen unbefriedigend. Die anschließende Analyse potenzieller Resistenzmechanismen zeigte, dass essentielle kostimulatorische Oberflächenmoleküle auf RMS fehlen, während Inhibitoren der Tumor T Zell Interaktion, wie B7H1 und B7H3, von RMS verstärkt exprimiert werden. Des Weiteren erwies sich die Expression von Adhäsionsmolekülen für die Effektivität chimärer Effektorzellen als relevant, da die CD54-Neoexpression auf mehreren primär CD54 negativen RMS Zelllinien die Zytotoxizität chimärer T und CIK Zellen in vitro signifikant steigerte. Als Weitere, teilweise RMS-Zelllinien-spezifische, potenzielle Resistenzmechanismen wurden stark verminderte Sensitivitäten gegenüber IFNγ, TNF, FAS-L und teilweise TRAIL gefunden. Außerdem zeigten RMS-Zelllinien und RMS-Gewebe eine starke Überexpression von Mitgliedern der inhibitor of apoptosis (IAP) Familie, die in die T-Zell-induzierte Apoptose eingreifen. Die siRNA vermittelte oder pharmakologische Hemmung von überexprimierten IAPs verbesserte die Zytotoxiziät chimärer T Zellen gegenüber RMS-Zellen signifikant. Erste Versuche mit immundefizienten NOD/SCID Mäusen zeigten, dass die einmalige Gabe fAChR reaktiver chimärer T Zellen das Wachstum von RMS Xenotransplantaten verlangsamen, die Tumorzellen aber nicht eradizieren konnte. Zukünftige In-vivo-Experimente haben daher das Ziel, die Ausschaltung der neu entdeckten Resistenzmechanismen mit der wiederholten Gabe verbesserter chimärer T Zellen zu verbinden, um die Effizienz des adoptiven Transfers RMS-spezifischer chimärer T-Zellen zu steigern

Stabilin-1 is expressed in human breast cancer and supports tumor growth in mammary adenocarcinoma mouse model

Stabilin-1 is a multifunctional scavenger receptor expressed on alternatively-activated macrophages. Stabilin-1 mediates phagocytosis of "unwanted-self" components, intracellular sorting, and endocytic clearance of extracellular ligands including SPARC that modulates breast cancer growth. The expression of stabilin-1 was found on tumor-associated macrophages (TAM) in mouse and human cancers including melanoma, lymphoma, glioblastoma, and pancreatic insulinoma. Despite its tumor-promoting role in mouse models of melanoma and lymphoma the expression and functional role of stabilin-1 in breast cancer was unknown. Here, we demonstrate that stabilin-1 is expressed on TAM in human breast cancer, and its expression is most pronounced on stage I disease. Using stabilin-1 knockout (ko) mice we show that stabilin-1 facilitates growth of mouse TS/A mammary adenocarcinoma. Endocytosis assay on stabilin-1 ko TAM demonstrated impaired clearance of stabilin-1 ligands including SPARC that was capable of inducing cell death in TS/A cells. Affymetrix microarray analysis on purified TAM and reporter assays in stabilin-1 expressing cell lines demonstrated no influence of stabilin-1 expression on intracellular signalling. Our results suggest stabilin-1 mediated silent clearance of extracellular tumor growth-inhibiting factors (e.g. SPARC) as a mechanism of stabilin-1 induced tumor growth. Silent clearance function of stabilin-1 makes it an attractive candidate for delivery of immunomodulatory anti-cancer therapeutic drugs to TAM

Pulmonary cancers across different histotypes share hybrid tuft cell/ionocyte-like molecular features and potentially druggable vulnerabilities

Tuft cells are chemosensory epithelial cells in the respiratory tract and several other organs. Recent studies revealed tuft cell-like gene expression signatures in some pulmonary adenocarcinomas, squamous cell carcinomas (SQCC), small cell carcinomas (SCLC), and large cell neuroendocrine carcinomas (LCNEC). Identification of their similarities could inform shared druggable vulnerabilities. Clinicopathological features of tuft cell-like (tcl) subsets in various lung cancer histotypes were studied in two independent tumor cohorts using immunohistochemistry (n = 674 and 70). Findings were confirmed, and additional characteristics were explored using public datasets (RNA seq and immunohistochemical data) (n = 555). Drug susceptibilities of tuft cell-like SCLC cell lines were also investigated. By immunohistochemistry, 10–20% of SCLC and LCNEC, and approximately 2% of SQCC expressed POU2F3, the master regulator of tuft cells. These tuft cell-like tumors exhibited “lineage ambiguity” as they co-expressed NCAM1, a marker for neuroendocrine differentiation, and KRT5, a marker for squamous differentiation. In addition, tuft cell-like tumors co-expressed BCL2 and KIT, and tuft cell-like SCLC and LCNEC, but not SQCC, also highly expressed MYC. Data from public datasets confirmed these features and revealed that tuft cell-like SCLC and LCNEC co-clustered on hierarchical clustering. Furthermore, only tuft cell-like subsets among pulmonary cancers significantly expressed FOXI1, the master regulator of ionocytes, suggesting their bidirectional but immature differentiation status. Clinically, tuft cell-like SCLC and LCNEC had a similar prognosis. Experimentally, tuft cell-like SCLC cell lines were susceptible to PARP and BCL2 co-inhibition, indicating synergistic effects. Taken together, pulmonary tuft cell-like cancers maintain histotype-related clinicopathologic characteristics despite overlapping unique molecular features. From a therapeutic perspective, identification of tuft cell-like LCNECs might be crucial given their close kinship with tuft cell-like SCLC

Canonical WNT/β-Catenin Signaling Plays a Subordinate Role in Rhabdomyosarcomas

The development of skeletal muscle from immature precursors is partially driven by canonical WNT/β-catenin signaling. Rhabdomyosarcomas (RMS) are immature skeletal muscle-like, highly lethal cancers with a variably pronounced blockade of muscle differentiation. To investigate whether canonical β-catenin signaling in RMS is involved in differentiation and aggressiveness of RMS, we analyzed the effects of WNT3A and of a siRNA-mediated or pharmacologically induced β-catenin knock-down on proliferation, apoptosis and differentiation of embryonal and alveolar RMS cell lines. While the canonical WNT pathway was maintained in all cell lines as shown by WNT3A induced AXIN expression, more distal steps including transcriptional activation of its key target genes were consistently impaired. In addition, activation or inhibition of canonical WNT/β-catenin only moderately affected proliferation, apoptosis or myodifferentiation of the RMS tumor cells and a conditional knockout of β-catenin in RMS of Ptchdel/+ mice did not alter RMS incidence or multiplicity. Together our data indicates a subordinary role of the canonical WNT/β-catenin signaling for RMS proliferation, apoptosis or differentiation and thus aggressiveness of this malignant childhood tumor

The state of the Martian climate

60°N was +2.0°C, relative to the 1981–2010 average value (Fig. 5.1). This marks a new high for the record. The average annual surface air temperature (SAT) anomaly for 2016 for land stations north of starting in 1900, and is a significant increase over the previous highest value of +1.2°C, which was observed in 2007, 2011, and 2015. Average global annual temperatures also showed record values in 2015 and 2016. Currently, the Arctic is warming at more than twice the rate of lower latitudes

Search for supersymmetry in events with b-quark jets and missing transverse energy in pp collisions at 7 TeV

Results are presented from a search for physics beyond the standard model based on events with large missing transverse energy, at least three jets, and at least one, two, or three b-quark jets. The study is performed using a sample of proton-proton collision data collected at sqrt(s) = 7 TeV with the CMS detector at the LHC in 2011. The integrated luminosity of the sample is 4.98 inverse femtobarns. The observed number of events is found to be consistent with the standard model expectation, which is evaluated using control samples in the data. The results are used to constrain cross sections for the production of supersymmetric particles decaying to b-quark-enriched final states in the context of simplified model spectra.Comment: Submitted to Physical Review

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements

Differential cross section measurements for the production of a W boson in association with jets in proton–proton collisions at √s = 7 TeV

Measurements are reported of differential cross sections for the production of a W boson, which decays into a muon and a neutrino, in association with jets, as a function of several variables, including the transverse momenta (pT) and pseudorapidities of the four leading jets, the scalar sum of jet transverse momenta (HT), and the difference in azimuthal angle between the directions of each jet and the muon. The data sample of pp collisions at a centre-of-mass energy of 7 TeV was collected with the CMS detector at the LHC and corresponds to an integrated luminosity of 5.0 fb[superscript −1]. The measured cross sections are compared to predictions from Monte Carlo generators, MadGraph + pythia and sherpa, and to next-to-leading-order calculations from BlackHat + sherpa. The differential cross sections are found to be in agreement with the predictions, apart from the pT distributions of the leading jets at high pT values, the distributions of the HT at high-HT and low jet multiplicity, and the distribution of the difference in azimuthal angle between the leading jet and the muon at low values.United States. Dept. of EnergyNational Science Foundation (U.S.)Alfred P. Sloan Foundatio

Penilaian Kinerja Keuangan Koperasi di Kabupaten Pelalawan

This paper describe development and financial performance of cooperative in District Pelalawan among 2007 - 2008. Studies on primary and secondary cooperative in 12 sub-districts. Method in this stady use performance measuring of productivity, efficiency, growth, liquidity, and solvability of cooperative. Productivity of cooperative in Pelalawan was highly but efficiency still low. Profit and income were highly, even liquidity of cooperative very high, and solvability was good