Christof Granzow

Synthesis of new boron-rich building blocks for boron neutron capture therapy or energy-filtering transmission electron microscopy.

The synthesis of a new ortho‐carborane derivative, tetracarboranylketone 4, is reported here. Ketone 4 was prepared from a tetraalkynylated ketone by the addition of decaborane. The keto group was then easily modified to yield the glycosides 17α and 18β, which contain glucose or galactose, respectively, and the nucleotide 13 b. In addition to ketone 4, which is acyclic, cyclic ketone 8 was also synthesised. X‐ray diffraction analysis of compound 4 indicated the presence of two toluene guest molecules per molecule of the host compound. Furthermore, compound 4 displays a rather low cytotoxicity. These novel products can be used as building blocks to create a new class of biomolecules containing high‐density carborane clusters. Such molecules may constitute powerful tools for applications like Boron Neutron Capture Therapy or Energy‐Filtering Transmission Electron Microscopy.

Abstract 1889: Formation and merger of H69 and H69AR small cell lung cancer (SCLC) tumor cell clusters (TCCs)in vitro

A novel manifestation of tumor cell invasiveness, termed mutual cellular pervasion (MCP), was observed in vitro. It involved TCCs formed by SCLC cells. Suspension cultures of H69 and H69AR cells were grown in flavin-free RPMI 1640 medium with 10% fetal bovine serum as the only source of flavins. Experiments and subcultivations were performed under light with wavelengths above 520 nm. TCC morphology was studied by confocal microscopy and transmission and scanning electron microscopy. Interactions of TCCs were followed by video microscopy in phase contrast and fluorescence modes. Data were analyzed using the Definiens software suite. The interaction of H69AR TCCs with confluent monolayers of human umbilical vascular endothelial cells (HUVEC) was visualized. Freshly subcultivated, singularized tumor cells rapidly formed aggregates. After two days, solid TCCs prevailed whose surface cells were connected by desmosome-like junctions. Apoptosis but not necrosis became obvious in central parts of clusters with diameters above 100 to 150 μm. TCCs enlarged through both cell proliferation and serial cluster mergers. In the course of TCC merger, four stages were distinguishable: (1) directed cluster movement leading to contact of clusters, (2) formation of a near circular, symmetric structure termed contact disc. It consists of planar surface areas to which both TCCs involved contribute equally. (3) Merger of TCCs took place in the contact disc, and was accompanied by active movement of adjacent cells. (4) Starting from the merger zone, major surface areas of newly formed clusters were mutually pervaded by cells from both participating cell clusters. After numerous merger events, only low numbers of TCCs reaching sizes of up to 2 to 3 mm were present at subcultivation. No spontaneous bursting, shrinkage, degeneration, or decay of TCCs has been observed. However, when TCCs of H69AR cells were confronted with confluent layers of HUVEC, drastic changes occurred in both formations. First, intense oscillatory cellular movements were triggered on TCC surfaces, preceding the release of cell aggregates which moved away from TCCs to the HUVEC layer. At the same time, HUVEC were disconnected from each other and started moving either randomly or in an oscillatory fashion. Individual tumor cells attached to the much larger HUVEC and spread on their surface, resulting in full covering which secluded HUVEC from the medium. To our knowledge, neither formation nor merger of malignant TCCs in vitro have been described so far. TCC merger involves mutual cellular penetration of the involved TCC surfaces, i. e. MCP. Contrary to the invasion of tumor cells into non-tumor tissue, pervasion does not lead to destruction but results in TCC growth. However, key features of tumor cell invasion in vivo, such as chemotaxis, spreading and TCC formation as well as disruption of endothelial layers, manifest under conditions in vitro, thus facilitating related experimental studies. Citation Format: Nico O. Ruprecht, Martin Hungerbuehler, Philipp Kellmann, Nathalie Harder, Gunter Schmidt, Carlos Wotzkow, Beat Haenni, Fabian Blank, Hendrik Von Tengg-Kobligk, Christof Granzow. Formation and merger of H69 and H69AR small cell lung cancer (SCLC) tumor cell clusters (TCCs) in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1889. doi:10.1158/1538-7445.AM2017-1889

Characterization of Proteins Involved in Mammalian Mitosis

Our interest in the molecular mechanisms of mitosis includes aspects of the mechanochemistry and fidelity of chromosome distribution, and the control of ontogeny which, in addition to differentiation, requires specific timing and positioning of cell divisions to generate the pattern of some 1014 cells of a mammal from a single fertilized egg and to compensate for cell losses.