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The FGFR4/FGF19 signaling axis is overactivated in 20% of liver tumors and currently represents a promising targetable signaling mechanism in this cancer type. However, blocking FGFR4 or FGF19 has proven challenging due to its physiological role in suppressing bile acid synthesis which leads to increased toxic bile acid plasma levels upon FGFR4 inhibition. An FGFR4-targeting antibody, U3-1784, was generated in order to investigate its suitability as a cancer treatment without major side effects.U3-1784 is a high-affinity fully human antibody that was obtained by phage display technology and specifically binds to FGFR4. The antibody inhibits cell signaling by competing with various FGFs for their FGFR4 binding site thereby inhibiting receptor activation and downstream signaling via FRS2 and Erk. The inhibitory effect on tumor growth was investigated in 10 different liver cancer models in vivo. The antibody specifically slowed tumor growth of models overexpressing FGF19 by up to 90% whereas tumor growth of models not expressing FGF19 was unaffected. In cynomolgus monkeys, intravenous injection of U3-1784 caused elevated serum bile acid and liver enzyme levels indicating potential liver damage. These effects could be completely prevented by the concomitant oral treatment with the bile acid sequestrant colestyramine, which binds and eliminates bile acids in the gut. These results offer a new biomarker-driven treatment modality in liver cancer without toxicity and they suggest a general strategy for avoiding adverse events with FGFR4 inhibitors.

The family of Mildew resistance Locus O (MLO) proteins is best known for its profound effect on the outcome of powdery mildew infections: when the appropriate MLO protein is absent, the plant is fully resistant to otherwise virulent powdery mildew fungi. However, most members of the MLO protein family remain functionally unexplored. Here, we investigateArabidopsis thaliana MLO3, the closest relative ofAtMLO2, AtMLO6andAtMLO12, which are the ArabidopsisMLOgenes implicated in the powdery mildew interaction. The co-expression network ofAtMLO3suggests association of the gene with plant defense-related processes such as salicylic acid homeostasis. Our extensive analysis shows thatmlo3mutants are unaffected regarding their infection phenotype upon challenge with the powdery mildew fungiGolovinomyces orontiiandErysiphe pisi, the oomyceteHyaloperonospora arabidopsidis, and the bacterial pathogenPseudomonas syringae(the latter both in terms of basal and systemic acquired resistance), indicating that the protein does not play a major role in the response to any of these pathogens. However,mlo3genotypes display spontaneous callose deposition as well as signs of early senescence in six-or seven-week-old rosette leaves in the absence of any pathogen challenge, a phenotype that is reminiscent ofmlo2mutant plants. We hypothesize that de-regulated callose deposition inmlo3genotypes is the result of a subtle transient aberration of salicylic acid-jasmonic acid homeostasis during development.

The journal of biological chemistry : JBC 296, 100611 (2021). doi:10.1016/j.jbc.2021.100611, Published by American Society for Biochemistry and Molecular Biology, Bethesda, MD.

Genes 11(5), 517 (2020). doi:10.3390/genes11050517, Published by MDPI, Basel

Barley mlo mutants are well known for their profound resistance against powdery mildew disease. Recently, mlo mutant plants were generated in hexaploid bread wheat (Triticum aestivum) with the help of transgenic (transcription-activator-like nuclease, TALEN) and non-transgenic (targeted induced local lesions in genomes, TILLING) biotechnological approaches. While full gene knockouts in the three wheat Mlo (TaMlo) homoeologs, created via TALEN, confer full resistance to the wheat powdery mildew pathogen (Blumeria graminis f.sp. tritici), the currently available TILLING-derived Tamlo missense mutants provide only partial protection against powdery mildew attack. Here we studied the infection phenotypes of TALEN- and TILLING-derived Tamlo plants to the two hemibiotrophic pathogens Zymoseptoria tritici, causing Septoria leaf blotch in wheat, and Magnaporthe oryzae pv. Triticum (MoT), the causal agent of wheat blast disease. While Tamlo plants showed unaltered outcomes upon challenge with Z. tritici, we found allele-specific levels of enhanced susceptibility to MoT, with stronger powdery mildew resistance correlated with more invasive growth by the blast pathogen. Surprisingly, unlike barley mlo mutants, young wheat mlo mutant plants do not show undesired pleiotropic phenotypes such as spontaneous callose deposits in leaf mesophyll cells or signs of early leaf senescence. In conclusion, our study provides evidence for allele-specific levels of enhanced susceptibility of Tamlo plants to the hemibiotrophic wheat pathogen MoT.

Human macrophage migration inhibitory factor (MIF) is an inflammatory cytokine with chemokine-like characteristics and an upstream regulator of host innate immunity. It is a critical mediator of a variety of human diseases, such as acute and chronic inflammatory diseases, autoimmunity, atherosclerosis, and cancer. MIF is an atypical chemokine that not only signals through its cognate receptor CD74, but also interacts with the classical chemokine receptors CXCR2 and CXCR4. MIF and its homolog D-dopachrome tautomerase (D-DT)/MIF-2 are structurally unique proteins that are conserved across kingdoms and that share a remarkable homology with bacterial tautomerases/isomerases, albeit the relevance of the tautomerase activity in mammalian systems has remained unclear. Intriguingly, in silico analysis also predicts MIF orthologs in plants such as in the model plant Arabidopsis thaliana. There are three predicted MIF orthologs in A. thaliana, which have been termed A. thaliana MIF/D-DT-like proteins (AtMDLs). Anticipating that there will be a future research interest in studying AtMDLs or other plant MDLs, here we describe methods how to clone, recombinantly express and purify AtMDL proteins, taking into account codon usage differences between plant and mammalian cell systems.

Human macrophage migration inhibitory factor (MIF) is an inflammatory cytokine with chemokine-like characteristics and an upstream regulator of host innate immunity. It is a critical mediator of a variety of human diseases, such as acute and chronic inflammatory diseases, autoimmunity, atherosclerosis, and cancer. MIF is an atypical chemokine that not only signals through its cognate receptor CD74, but also interacts with the classical chemokine receptors CXCR2 and CXCR4. MIF and its homolog D-dopachrome tautomerase (D-DT)/MIF-2 are structurally unique proteins that are conserved across kingdoms and that share a remarkable homology with bacterial tautomerases/isomerases, albeit the relevance of the tautomerase activity in mammalian systems has remained unclear. Intriguingly, in silico analysis also predicts MIF orthologs in plants such as in the model plant Arabidopsis thaliana. There are three predicted MIF orthologs in A. thaliana, which have been termed A. thaliana MIF/D-DT-like proteins (AtMDLs). Anticipating that there will be a future research interest in studying AtMDLs or other plant MDLs, here we describe methods how to clone, recombinantly express and purify AtMDL proteins, taking into account codon usage differences between plant and mammalian cell systems.

Members of the MILDEW RESISTANCE LOCUS O (MLO) gene family confer susceptibility to powdery mildews in different plant species, and their existence therefore seems to be disadvantageous for the plant. We recognized that expression of the Arabidopsis MLO2 gene is induced after inoculation with the bacterial pathogen Pseudomonas syringae, promoted by salicylic acid (SA) signaling, and systemically enhanced in the foliage of plants exhibiting systemic acquired resistance (SAR). Importantly, distinct mlo2 mutant lines were unable to systemically increase resistance to bacterial infection after inoculation with P. syringae, indicating that the function of MLO2 is necessary for biologically induced SAR in Arabidopsis. Our data also suggest that the close homolog MLO6 has a supportive but less critical role in SAR. In contrast to SAR, basal resistance to bacterial infection was not affected in mlo2. Remarkably, SAR-defective mlo2 mutants were still competent in systemically increasing the levels of the SAR-activating metabolites pipecolic acid (Pip) and SA after inoculation, and to enhance SAR-related gene expression in distal plant parts. Furthermore, although MLO2 was not required for SA- or Pip-inducible defense gene expression, it was essential for the proper induction of disease resistance by both SAR signals. We conclude that MLO2 acts as a critical downstream component in the execution of SAR to bacterial infection, being required for the translation of elevated defense responses into disease resistance. Moreover, our data suggest a function for MLO2 in the activation of plant defense priming during challenge by P. syringae.

Human macrophage migration-inhibitory factor (MIF) is an evolutionarily-conserved protein that has both extracellular immune-modulating and intracellular cell-regulatory functions. MIF plays a role in various diseases, including inflammatory diseases, atherosclerosis, autoimmunity, and cancer. It serves as an inflammatory cytokine and chemokine, but also exhibits enzymatic activity. Secreted MIF binds to cell-surface immune receptors such as CD74 and CXCR4. Plants possess MIF orthologs but lack the associated receptors, suggesting functional diversification across kingdoms. Here, we characterized three MIF orthologs (termed MIF/d-dopachrome tautomerase–like proteins or MDLs) of the model plantArabidopsis thaliana. RecombinantArabidopsisMDLs (AtMDLs) share similar secondary structure characteristics with human MIF, yet only have minimal residual tautomerase activity using eitherp-hydroxyphenylpyruvate or dopachrome methyl ester as substrate. Site-specific mutagenesis suggests that this is due to a distinct amino acid difference at the catalytic cavity-defining residue Asn-98. Surprisingly,AtMDLs bind to the human MIF receptors CD74 and CXCR4. Moreover, they activate CXCR4-dependent signaling in a receptor-specific yeast reporter system and in CXCR4-expressing human HEK293 transfectants. Notably, plant MDLs exert dose-dependent chemotactic activity toward human monocytes and T cells. A small molecule MIF inhibitor and an allosteric CXCR4 inhibitor counteract this function, revealing its specificity. Our results indicate cross-kingdom conservation of the receptor signaling and leukocyte recruitment capacities of human MIF by its plant orthologs. This may point toward a previously unrecognized interplay between plant proteins and the human innate immune system.

The FGFR4/FGF19 signaling axis is overactivated in 20% of liver tumors and currently represents a promising targetable signaling mechanism in this cancer type. However, blocking FGFR4 or FGF19 has proven challenging due to its physiological role in suppressing bile acid synthesis which leads to increased toxic bile acid plasma levels upon FGFR4 inhibition. An FGFR4-targeting antibody, U3-1784, was generated in order to investigate its suitability as a cancer treatment without major side effects. U3-1784 is a high-affinity fully human antibody that was obtained by phage display technology and specifically binds to FGFR4. The antibody inhibits cell signaling by competing with various FGFs for their FGFR4 binding site thereby inhibiting receptor activation and downstream signaling via FRS2 and Erk. The inhibitory effect on tumor growth was investigated in 10 different liver cancer models in vivo. The antibody specifically slowed tumor growth of models overexpressing FGF19 by up to 90% whereas tumor growth of models not expressing FGF19 was unaffected. In cynomolgus monkeys, intravenous injection of U3-1784 caused elevated serum bile acid and liver enzyme levels indicating potential liver damage. These effects could be completely prevented by the concomitant oral treatment with the bile acid sequestrant colestyramine, which binds and eliminates bile acids in the gut. These results offer a new biomarker-driven treatment modality in liver cancer without toxicity and they suggest a general strategy for avoiding adverse events with FGFR4 inhibitors.

We investigated the relationships of the two immune-regulatory plant metabolites, salicylic acid (SA) and pipecolic acid (Pip), in the establishment of plant systemic acquired resistance (SAR), SAR-associated defense priming, and basal immunity. Using SA-deficient sid2, Pip-deficient ald1, and sid2 ald1 plants deficient in both SA and Pip, we show that SA and Pip act both independently from each other and synergistically in Arabidopsis thaliana basal immunity to Pseudomonas syringae. Transcriptome analyses reveal that SAR establishment in Arabidopsis is characterized by a strong transcriptional response systemically induced in the foliage that prepares plants for future pathogen attack by preactivating multiple stages of defense signaling and that SA accumulation upon SAR activation leads to the downregulation of photosynthesis and attenuated jasmonate responses systemically within the plant. Whereas systemic Pip elevations are indispensable for SAR and necessary for virtually the whole transcriptional SAR response, a moderate but significant SA-independent component of SAR activation and SAR gene expression is revealed. During SAR, Pip orchestrates SA-dependent and SA-independent priming of pathogen responses in a FLAVIN-DEPENDENT-MONOOXYGENASE1 (FMO1)-dependent manner. We conclude that a Pip/FMO1 signaling module acts as an indispensable switch for the activation of SAR and associated defense priming events and that SA amplifies Pip-triggered responses to different degrees in the distal tissue of SAR-activated plants.

Genome-wide microarray analyses revealed that during biological activation of systemic acquired resistance (SAR) in Arabidopsis, the transcript levels of several hundred plant genes were consistently up- (SAR(+) genes) or down-regulated (SAR(-) genes) in systemic, non-inoculated leaf tissue. This transcriptional reprogramming fully depended on the SAR regulator FLAVIN-DEPENDENT MONOOXYGENASE1 (FMO1). Functional gene categorization showed that genes associated with salicylic acid (SA)-associated defenses, signal transduction, transport, and the secretory machinery are overrepresented in the group of SAR(+) genes, and that the group of SAR(-) genes is enriched in genes activated via the jasmonate (JA)/ethylene (ET)-defense pathway, as well as in genes associated with cell wall remodeling and biosynthesis of constitutively produced secondary metabolites. This suggests that SAR-induced plants reallocate part of their physiological activity from vegetative growth towards SA-related defense activation. Alignment of the SAR expression data with other microarray information allowed us to define three clusters of SAR(+) genes. Cluster I consists of genes tightly regulated by SA. Cluster II genes can be expressed independently of SA, and this group is moderately enriched in H2O2- and abscisic acid (ABA)-responsive genes. The expression of the cluster III SAR(+) genes is partly SA-dependent. We propose that SA-independent signaling events in early stages of SAR activation enable the biosynthesis of SA and thus initiate SA-dependent SAR signaling. Both SA-independent and SA-dependent events tightly co-operate to realize SAR. SAR(+) genes function in the establishment of diverse resistance layers, in the direct execution of resistance against different (hemi-)biotrophic pathogen types, in suppression of the JA- and ABA-signaling pathways, in redox homeostasis, and in the containment of defense response activation. Our data further indicated that SAR-associated defense priming can be realized by partial pre-activation of particular defense pathways.

Fibroblast Growth Factor Receptor 4 (FGFR4) is the fourth member of the Fibroblast Growth Factor Receptor (FGFR) family of receptor tyrosine kinases. All of the FGFR's have been implicated in cancer development due to increased activation of their enzymatic activity either by gene mutation, over-expression or inadvertent ligand-mediated stimulation. One important alteration that may lead to FGFR4 activation in cancer is the overexpression of its ligand FGF19 in 20-40% of primary liver cancer. Here, we report the development of U3-1784, a phage display-derived fully human antibody that specifically binds to FGFR4 but not to isoforms of FGFR1-3. The antibody binds to an epitope in the putative ligand binding domain of the receptor and consequently inhibits ligand binding and downstream signaling. In a panel of 10 tumor models derived from hepatocellular carcinoma, U3-1784 significantly inhibits the growth of FGF19-expressing models up to 90% whereas models without FGF19 expression are insensitive. These results strongly suggest that the FGFR4/FGF19 axis is an oncogenic driver in hepatocellular carcinoma. U3-1784 is currently in phase I clinical trials. Citation Format: Rene Bartz, Keisuke Fukuchi, Tanja Lange, Katrin Gruner, Toshiaki Ohtsuka, Ichiro Watanabe, Shinko Hayashi, Mauricio Redondo-Müller, Mizuki Takahashi, Toshinori Agatsuma, Johannes Bange, Reimar Abraham. U3-1784, a human anti-FGFR4 antibody for the treatment of cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3852.

Ein Unternehmen ohne eine fundierte Unternehmensstrategie ist in der heutigen durch eine hohe Wettbewerbsdynamik gekennzeichneten wirtschaftlichen Orientierung und Marktlage nahezu undenkbar. In der Blutezeit des E-Business (1999–2000) ist eine strategische Ausrichtung sich neu grundender Unternehmen ganzlich vernachlassigt worden. Viele Start-Up’s sind davon ausgegangen, dass die Veroffentlichung einer oder mehrerer Internetseiten mit einer entsprechenden Verkaufsfunktion als strategische Orientierung eines Unternehmens ausreicht. Diese Quick and Dirty Theorie wurde durch die Etablierung des Neuen Marktes und dem damit einsetzenden Borsenboom untermauert, so dass eines der wichtigsten geschaftlichen Grundprinzipien, die langfristige Orientierung eines Unternehmens mit einer detaillierten Ausrichtung nach dem okonomischen Prinzip, auser Kraft gesetzt worden ist. Essenziell fur ein neu gegrundetes Unternehmen war die Tatsache, dass es sich schnell namentlich in der Offentlichkeit bekannt machte (der sogenannte First-MoverEffekt). Letztendlich hat sich fur diese Unternehmen das Sprichwort nach Konfuzius „Wer nicht an die Zukunft denkt, der wird bald grose Sorgen haben“ bewahrheitet.

Bevor ein Controlling / ein IT-Controlling die Wertschopfungskette von IT-Projekten mit effizienten Controlling-Instrumentarien unterstutzen kann, ist es erforderlich, die zu „controllende“ Wertschopfungskette naher zu spezifizieren. Nur wenn eine Wegbeschreibung, sei sie auch noch so grob strukturiert, vorliegt, lasst sich ein Ziel erreichen. Bezuglich der Wertschopfungskette von IT-Projekten bedeutet dies, dass diese in ihren Grundstrukturen dem Betrachter bekannt ist, bevor er effektiv und effizient arbeiten kann. Daher widmet sich dieses Kapitel im Speziellen dieser Anforderung, um hier die Grundlagen fur die weiteren Ausfuhrungen zu legen.

Nach Abschluss eines IT-Projektes wird das entsprechende Ergebnis dem Auftraggeber ubergeben, der dieses IT-Produkt im Sinne seiner Unternehmensprodukte einsetzt. Es wird die letzte Stufe der Wertschopfungskette von Software-Produkten angestosen, in der das entwickelte Produkt einen positiven Ergebnisbeitrag fur das Unternehmen zu erbringen hat. Gleichzeitig erfolgt auch eine Analyse der abgeschlossenen Projekte, um Erkenntnisse und Verbesserungspotenzial fur nachfolgende Projekte zu ermitteln. Die Ergebnisse dieser Analyse sind in einer Projekterfahrungsdatenbank festzuhalten, die jedem Projektmitarbeiter respektive dem Controlling zuganglich ist. Insbesondere gilt es, die Periodenberichte unter dem Aspekt des Aufwandsanfalles zu untersuchen. So ist festzustellen, in welchen Perioden oder auch Software-Entwicklungsphasen der Projektaufwand gegenuber den Planwerten erhoht gewesen ist und ob man dort mit verbesserten Methoden und Werkzeugen Einsparungen hatte erzielen konnen.

Nach Definition der Unternehmensstrategie und der daraus abgeleiteten Strategie fur den IT-Bereich tritt die nachste Phase innerhalb des Software-Produktlebenszyklusses in Kraft. Strategien sind lediglich Handlungspramissen, die ohne weitere Aktivitaten nicht umsetzbar sind. Durch die Verwendung der Balanced Scorecard Methode sind diese zwar teilweise zu konkreten Handlungsempfehlungen ausgebaut worden, doch stehen vielfach noch konkrete Masnahmen zu deren Umsetzung aus. So ist es nicht ratsam, ein Strategiegedanke ohne jegliche Wirtschaftlichkeitsbetrachtungen beziehungsweise Detailanalysen bezuglich dessen Praxistauglichkeit in die Realitat umzusetzen. Des Weiteren sind auch entsprechende geldwerte Mittel fur den Umsetzungsprozess bereitzustellen, die in jedem Unternehmen als knappes Gut angesehen werden. Es gilt diese geldwerten Mittel unter dem Primat der unternehmensbezogenen Wertsteigerung zielgerichtet zu verteilen. Hierzu ist die Aufstellung eines Budgetplans erforderlich, der die erwarteten Ausgaben und/oder Einnahmen auf der Grundlage der genauesten verfugbaren Informationen bestimmter Kosten- und Einnahmearten fur eine definierte Zeit enthalt214.

Die Bedeutung eines effizienten und effektiven Controllings innerhalb eines Unternehmens wird an dieser Stelle nicht naher erlautert. Sie wird als hoch vorausgesetzt, da das Controlling die Aufgabe hat, als Fuhrungssubsystem die Unternehmensfuhrung zu unterstutzen94. Des Weiteren obliegt ihm die Aufgabe, die Aktivitaten aller Unternehmensteilbereiche sowohl in sachlicher als auch zeitlicher Hinsicht im Sinne der Unternehmensziele abzustimmen. Durch gezielte Plan-Ist-Vergleiche werden Abweichungen aufgezeigt, analysiert und kommuniziert, sowie Masnahmen zur Gegensteuerung vorgeschlagen, teilweise selbstandig eingeleitet und koordiniert95. Hierzu ist die Entwicklung unternehmensspezifischer Planungs- und Planungskontrollsysteme erforderlich, die dem Controlling ein zukunftsorientiertes Denken und Handeln ermoglichen96. Als funktionsubergreifendes Steuerungsinstrumentarium sorgt es sich um eine ergebnisorientierte Koordination von Planungs-, Kontroll- und Informationsversorgungsmechanismen97, die erst eine nach dem okonomischen Prinzip verfahrende Unternehmenssteuerung erlauben.

IT-Vorhaben werden in der Regel in Projektform abgewickelt. Dieser Abwicklungsvorgang reprasentiert meistens nur die operative Umsetzung einer Idee beziehungsweise einer expliziten Anforderung. Jedes Unternehmen strebt nach einem geldwerten okonomischen Erfolg, der durch die Erfullung von Kundenbedurfnissen erlangt wird. Ein Projekt legt hierfur im IT-Bereich die Grundlagen durch Schaffung eines entsprechenden IT-Produktes. Es kann jedoch nur einen Teilprozess eines komplexen Gefuges verkorpern, das dem Ziel der Befriedigung eines oder mehrerer Kundenbedurfnisse dient.