Tyrosine kinase receptor B (TrkB) expression in colorectal cancers highlights anoikis resistance as a survival mechanism of tumour budding cells.

AIMS Tumour buds in colorectal cancer represent an aggressive subgroup of non-proliferating and non-apoptotic tumour cells. We hypothesize that the survival of tumour buds is dependent upon anoikis resistance. The role of tyrosine kinase receptor B (TrkB), a promoter of epithelial-mesenchymal transition and anoikis resistance, in facilitating budding was investigated. METHODS AND RESULTS Tyrosine kinase receptor B immunohistochemistry was performed on a multiple-punch tissue microarray of 211 colorectal cancer resections. Membranous/cytoplasmic and nuclear expression was evaluated in tumour and buds. Tumour budding was assessed on corresponding whole tissue slides. Relationship to Ki-67 and caspase-3 was investigated. Analysis of Kirsten Ras (KRAS), proto-oncogene B-RAF (BRAF) and cytosine-phosphate-guanosine island methylator phenotype (CIMP) was performed. Membranous/cytoplasmic and nuclear TrkB were strongly, inversely correlated (P < 0.0001; r = -0.41). Membranous/cytoplasmic TrkB was overexpressed in buds compared to the main tumour body (P < 0.0001), associated with larger tumours (P = 0.0236), high-grade budding (P = 0.0011) and KRAS mutation (P = 0.0008). Nuclear TrkB was absent in buds (P <0.0001) and in high-grade budding cancers (P =0.0073). Among patients with membranous/cytoplasmic TrkB-positive buds, high tumour membranous/cytoplasmic TrkB expression was a significant, independent adverse prognostic factor [P = 0.033; 1.79, 95% confidence interval (CI) 1.05-3.05]. Inverse correlations between membranous/cytoplasmic TrkB and Ki-67 (r = -0.41; P < 0.0001) and caspase-3 (r =-0.19; P < 0.05) were observed. CONCLUSIONS Membranous/cytoplasmic TrkB may promote an epithelial-mesenchymal transition (EMT)-like phenotype with high-grade budding and maintain viability of buds themselves.

VEGFR2 Signaling Prevents Colorectal Cancer Cell Senescence to Promote Tumorigenesis in Mice With Colitis.

BACKGROUND & AIMS Senescence prevents cellular transformation. We investigated whether vascular endothelial growth factor (VEGF) signaling via its receptor, VEGFR2, regulates senescence and proliferation of tumor cells in mice with colitis-associated cancer (CAC). METHODS CAC was induced in VEGFR2(ΔIEC) mice, which do not express VEGFR2 in the intestinal epithelium, and VEGFR2(fl/fl) mice (controls) by administration of azoxymethane followed by dextran sodium sulfate. Tumor development and inflammation were determined by endoscopy. Colorectal tissues were collected for immunoblot, immunohistochemical, and quantitative polymerase chain reaction analyses. Findings from mouse tissues were confirmed in human HCT116 colorectal cancer cells. We analyzed colorectal tumor samples from patients before and after treatment with bevacizumab. RESULTS After colitis induction, VEGFR2(ΔIEC) mice developed significantly fewer tumors than control mice. A greater number of intestinal tumor cells from VEGFR2(ΔIEC) mice were in senescence than tumor cells from control mice. We found VEGFR2 to activate phosphatidylinositol-4,5-bisphosphate-3-kinase and AKT, resulting in inactivation of p21 in HCT116 cells. Inhibitors of VEGFR2 and AKT induced senescence in HCT116 cells. Tumor cell senescence promoted an anti-tumor immune response by CD8(+) T cells in mice. Patients whose tumor samples showed an increase in the proportion of senescent cells after treatment with bevacizumab had longer progression-free survival than patients in which the proportion of senescent tumor cells did not change before and after treatment. CONCLUSIONS Inhibition of VEGFR2 signaling leads to senescence of human and mouse colorectal cancer cells. VEGFR2 interacts with phosphatidylinositol-4,5-bisphosphate-3-kinase and AKT to inactivate p21. Colorectal tumor senescence and p21 level correlate with patient survival during treatment with bevacizumab.

Reactions of CF3-enones with arenes under superelectrophilic activation: a pathway to trans-1,3-diaryl-1-CF3-indanes, new cannabinoid receptor ligands

4-Aryl-1,1,1-trifluorobut-3-en-2-ones ArCH[double bond, length as m-dash]CHCOCF3 (CF3-enones) react with arenes in excess of Brønsted superacids (TfOH, FSO3H) to give, stereoselectively, trans-1,3-diaryl-1-trifluoromethyl indanes in 35-85% yields. The reaction intermediates, the O-protonated ArCH[double bond, length as m-dash]CHC(OH(+))CF3 and the O,C-diprotonated ArHC(+)CH2C(OH(+))CF3 species, have been studied by means of (1)H, (13)C, (19)F NMR, and DFT calculations. Both types of the cations may participate in the reaction, depending on their electrophilicity and electron-donating properties of the arenes. The formation of CF3-indanes is a result of cascade reaction of protonated CF3-enones to form chemo-, regio- and stereoselectively three new C-C bonds. The obtained trans-1,3-diaryl-1-trifluoromethyl indanes were investigated as potential ligands for cannabinoid receptors CB1 and CB2 types. The most potent compound showed sub-micromolar affinity for both receptor subtypes with a 6-fold selectivity toward the CB2 receptor with no appreciable cytotoxicity toward SHSY5Y cells.

Na 1 Nachlass Max Horkheimer, 640 - "Simmel and Freudian Philosophy", "Authoritarianism and the Family Today" und "The Chances of Democracy in Germany" (p. IX 18.1-3-IX 19a)

"Simmel and Freudian Philosophy" (GS 5, S. 396-405); 1. Nachruf, verlesen beim Memorial Meeting for Ernst Simmel; datiert: 13.12.1947; veröffentlicht in: International Journal of Psychoanalysis, 29. Jahrgang, 1948, S. 110-113; 2. Abschrift aus Werken und Briefen Siegmund Freuds; Typoskript, 9 Blatt; 3. Freeman, Burriel: 1 Brief mit Unterschirft an Max Horkheimer, Chicago, 10.06.1949; 1 Brief von Max Horkheimer, Los Angeles, 15.06.1949, 2 Blatt; "Authoritarianism and the Family Today" (GS 5, S. 377-395); 1. Aufsatz, datiert 1947, veröffentlicht in: Ruth Nanda Anshen (editor), "The Family: Its Function and Distiny", New York 1949. a) Typsokript, 20 Blatt b) Typoskript mit handschriftlichen Korrekturen, 20 Blatt c) Typoskript mit eigenhändigen Korrekturen, 20 Blatt d)-f) deutsche Fassung mit dem Titel "Autorität und Familie", übersetzt vom Institut für Sozialforschung, 1960; veröffentlicht in : "Erkenntnis und Verantwortung. Festschrift für Theodor Litt", Düsseldorft, 1960 d) Typoskript, 20 Blatt e) Typoskript, 20 Blatt f) Korrekturfahnen aus der Litt- Festschrift, mit dem Titel "Autorität und Familie in der Gegenwart"; 6 Blatt; 2. Schönbach, Peter: 1 Brief mit Unterschrift an Max Horkheier, ohne Ort, 23.06.1960; 1 Blatt; 3. Schönbach, Peter: 1 Brief mit Unterschrift an Friedrich Pollock, ohne Ort, 22.06.1960; 1 Blatt; "The Chances of Democracy in Germany" (GS 12, S. 184-194); 1947 [?] a) Typoskript, 10 Blatt b) Typoskript mit eigenhändigen Korrekturen, 11 Blatt c) Typoskript mit eigenhändigen Korrekturen ,11 Blatt;

Na 1 Nachlass Max Horkheimer, 712 - Entwürfe für ein mögliches Forschungsprojekt über Liberalismus des 19. Jahrhunderts; "Untersuchung des sozialen Klimas in Stadt- und Landkreisen Hessens"; "Betriebsklime. Eine industrie-soziologische Untersuchung aus dem Ruhrgebiet"; Sardemann, Karl: Interview mit einem Juden über sein Leben 1933 bis circa 1953; "Altersbild und Altersvorsorge der Arbeiter und Angestellte" (p. IX 187 - IX 191.1-3)

Entwürfe für ein Mögliches Forschungsprojekt über Liberlaismus des 19. Jahrhunderts; 1953; 1. Typoskript mit eigenhändigen Korrekturen von Theodor W. Adorno; 7 Blatt; 2. Typoskript mit eigenhändigen Korrekturen von Theodor W. Adorno; 3 Blatt; "Untersuchungn des sozialen Klimas in Stadt- und Landkreisen Hessens"; Entwürfe für ein "Handbuch der Methoden zur Messung des sozialen Klimas", nicht veröffentlicht; 1953; 1. Pollock, Friedrich: "Einführung" zum "Handbuch" a) Typoskript mit eigenhändigen Korrekturen; 8 Blatt; b) Typoskript mit eigenhändigen Korrekturen, 5 Blatt; c) Manuskript, "Vorwort"; 5 Blatt; d) Baumert, Gerhard: Entwurf des "Vorworts"; Typoskript mit handschriftlichen Korrekturen, 2 Blatt; e) Baumert, Gerhard: "Notizen zu Vorwort und Einführung des Handbuchs"; Typoskript, 1 Blatt; 2. Inhaltsverzeichnis zum "Handbuch"; 1 Blatt; 3. Pollock, Friedrich: eigenhändige Notizen zur Einführung; 7 Blatt; 4. Pollock, Friedrich: 1 eigenhändiger Brief mit Unterschrift an Theodor W. Adorno, Santa Monica, 21.09.1953; 1 Blatt; "Betriebsklime. Eine industrie-soziologische Untersuchung aus dem Ruhrgebiet" 1954-1956 veröffentlicht als Band 3 der Frankfurter Beiträge zur Soziologie, Frankfurt 1955; 1. Druckfahnen, mit handschriftlichen Korrekturen; 38 Blatt; 2. Korrektur-Notizen zu den Druckfahnen; 4 Blatt; 3. Adorno, Theodor W. [Mitarbeit]: "Grundreiz [Betriebsumfrage)" a) Typoskript mit handschriftlichen Korrekturen, 4 Blatt; b) Typoskript, 5 Blatt; 4. Adorno, Theodor W.: 1 Brief an Max Horkheimer, ohne Ort, 30.06.1954; 1 Blatt; 5. Fragebogen- Entwurf; Typsokript, 2 Blatt; 6. Fragebogen- Entwurf; Typoskript, 2 Blatt; 7. Adorno, Theodor W.: 1 Brief an Max Horkheimer, ohne Ort, ohne Datum [1954]; 1 Blatt; 8. Dirks, Walter: "Notiz über meine Reise nach Köln, Düsseldorf und Essen", 22.02.1955. Typoskript, 2 Blatt; 9. Becker, Hellmut: "Aktennotiz, Betreff: Ersetzung der Pressekonferenz über die Mannesmann-Studie durch in den nächsten Wochen und Monaten aufeinanderfolgende Artikel qualifizierter Korrespondenten", 15.02.1955. Typoskript, 1 Blatt; 10. Adorno, Theodor W.: 1 Brief an Hermann Winkhaus, Mannesmann AG, ohne Ort, 05.03.1956; 2 Blatt; 11. Winkhaus, Hermann, Mannesmann AG: 1 Breifabschrift an das Institut für Sozialforschung, Düsseldorf, 18.07.1955; 12. Presseveröffentlichungen zur Betriebsklima-Untersuchung; 1 Ordner, 13 Blatt; 13. Zeitschriftenartikel und Abschriften von Artikeln zur Betriebsklima- Untersuchung; mit: Becker, Egon: 2 Briefe mit Unterschrift an Max Horkheimer, Frankfurt, 1955; 1 Brief von Max Horkheimer, Zürich, 25.10.1955; Sardemann, Karl: Interview mit einem Juden über sein Leben 1933 bis circa 1953, insbesondere seine Erlebnisse im Konzentrationslager; 1955; 1. Interview; Typsokript, 38 Blatt; 2. Sardemann, Karl: 1 Brief mit Unterschrift an Max Horkheimer, ohne Ort, 15.12.1955; 1 Blatt; "Altersbild und Altersvorsorge der Arbeiter und Angestellten" veröffentlicht als Sonderheft 1 der Frankfurter Beiträge zur Soziologie, Frankfurt 1958.; Zwischenbericht zum Projekt und Material, 1955; 1. Becker, Egon, u.a.: "Zwischenbericht" 10.03.1953; Typoskript, 9 Blatt; mit einem Brief mit Unterschrift von Egon Becker an Max Horkheimer, ohne Ort, 11.03.1955; 1 Blatt; 2. Fragebogen; als Typoskript vervielfältigt, 16 Blatt; 3. Fragebogen; als Typoskript vervielfältigt, 11 Blatt;

Oncogenic activation of c-Abl tyrosine kinase in non-small cell lung cancer: FUS1 tumor suppressor down-regulates c-Abl tyrosine kinase

The FUS1 tumor suppressor gene (TSG) has been found to be deficient in many human non-small cell lung cancer (NSCLC) tissue samples and cell lines (1,2,3). Studies have shown potent anti-tumor activity of FUS1 in animal models where FUS1 was delivered through a liposomal vector (4) and the use of FUS1 as a therapeutic agent is currently being studied in clinical human trials (5). Currently, the mechanisms of FUS1 activity are being investigated and my studies have shown that c-Abl tyrosine kinase is inhibited by the FUS1 TSG.^ Considering that many NSCLC cell lines are FUS1 deficient, my studies further identified that FUS1 deficient NSCLC cells have an activated c-Abl tyrosine kinase. C-Abl is a known proto-oncogene and while c-Abl kinase is tightly regulated in normal cells, constitutively active Abl kinase is known to contribute to the oncogenic phenotype in some types of hematopoietic cancers. My studies show that the active c-Abl kinase contributes to the oncogenicity of NSCLC cells, particularly in tumors that are deficient in FUS1, and that c-Abl may prove to be a viable target in NSCLC therapy.^ Current studies have shown that growth factor receptors play a role in NSCLC. Over-expression of the epidermal growth factor receptor (EGFR) plays a significant role in aggressiveness of NSCLC. Current late stage treatments include EFGR tyrosine kinase inhibitors or EGFR antibodies. Platelet-derived growth factor receptor (PDGFR) also has been shown to play a role in NSCLC. Of note, both growth factor receptors are known upstream activators of c-Abl kinase. My studies indicate that growth factor receptor simulation along deficiency in FUS1 expression contributes to the activation of c-Abl kinase in NSCLC cells. ^

Circumvention of cellular defense responses to DNA -directed nucleoside analogues by the protein kinase inhibitor, UCN-01

DNA-directed nucleoside analogues, such as ara-C, fludarabine, and gemcitabine, are antimetabolites effective in the treatment of a variety of cancers. However, resistance to nucleoside analogue-based chemotherapy in treatments is still a major problem in therapy. Therefore, it is essential to develop rationales for optimizing the use of nucleoside analogues in combination with other anticancer drugs or modalities such as radiation. The present study focuses on establishing mechanism-based combination strategy to overcome resistance to nucleoside analogues. ^ I hypothesized that the cytostatic concentrations of nucleoside analogues may cause S-phase arrest by activating an S-phase checkpoint that consists of a series of kinases. This may allow cells to repair damaged DNA over time and spare cytotoxicity. Thus, the ability of cells to enact an S-phase arrest in response to incorporation of potentially lethal amounts of nucleoside analogue may serve as a mechanism of resistance to S-phase-specific agents. As a corollary, the addition of a kinase inhibitor, such as UCN-01, may dysregulate the checkpoint response and abrogate the survival of S-phase-arrested cells by suppression of the survival signaling pathways. Using gemcitabine as a model of S-phase-specific nucleoside analogues in human acute myelogenous leukemia ML-1 cells, I demonstrated that cells arrested in S-phase in response to cytostatic conditions. Proliferation continued after washing the cells into drug-free medium, suggesting S-phase arrest served as a resistance mechanism of cancer cells to spare cytotoxicity of nucleoside analogues. However, nontoxic concentrations of UCN-01 rapidly killed S-phase-arrested cells by apoptosis. Furthermore, the molecular mechanism for UCN-01-induced apoptosis in S-phase-arrested cells was through inhibition of survival pathways associated with these cells. In this regard, suppression of the PI 3-kinase-Akt-Bad survival pathway as well as the NF-κB signaling pathway were associated with induction of apoptosis in S-phase-arrested cells by UCN-01, whereas the Ras-Raf-MEK-ERK pathway appeared not involved. This study has provided the rationales and strategies for optimizing the design of effective combination therapies to overcome resistance to nucleoside analogues. In fact, a clinical trial of the combination of ara-C with UCN-01 to treat relapsed or refractory AML patients has been initiated at U.T.M.D. Anderson Cancer Center. ^