Thomas Geiger

A Twist-Snail Axis Critical for TrkB-Induced Epithelial-Mesenchymal Transition-Like Transformation, Anoikis Resistance, and Metastasis

ABSTRACT In a genomewide anoikis suppression screen for metastasis genes, we previously identified the neurotrophic receptor tyrosine kinase TrkB. In mouse xenografts, activated TrkB caused highly invasive and metastatic tumors. Here, we describe that TrkB also induces a strong morphological transformation, resembling epithelial-mesenchymal transition (EMT). This required TrkB kinase activity, a functional mitogen-activated protein kinase pathway, suppression of E-cadherin, and induction of Twist, a transcription factor contributing to EMT and metastasis. RNA interference (RNAi)-mediated Twist depletion blocked TrkB-induced EMT-like transformation, anoikis suppression, and growth of tumor xenografts. By searching for essential effectors of TrkB-Twist signaling, we found that Twist induces Snail, another EMT regulator associated with poor cancer prognosis. Snail depletion impaired EMT-like transformation and anoikis suppression induced by TrkB, but in contrast to Twist depletion, it failed to inhibit tumor growth. Instead, Snail RNAi specifically impaired the formation of lung metastases. Epistasis experiments suggested that Twist acts upstream from Snail. Our results demonstrate that TrkB signaling activates a Twist-Snail axis that is critically involved in EMT-like transformation, tumorigenesis, and metastasis. Moreover, our data shed more light on the epistatic relationship between Twist and Snail, two key transcriptional regulators of EMT and metastasis.

Critical Role for TrkB Kinase Function in Anoikis Suppression, Tumorigenesis, and Metastasis

Anoikis, or cell death induced by cell detachment, provides protection against the metastatic spread of tumor cells. We have previously shown that the neurotrophic receptor tyrosine kinase TrkB suppresses anoikis in rat intestinal epithelial cells and renders them highly tumorigenic and metastatic. Because TrkB is overexpressed in several aggressive human cancers, first attempts are being made to target TrkB in cancer therapy. However, the mechanisms underlying TrkB-mediated anoikis suppression, tumorigenesis, and metastasis still remain largely elusive. Although, to date, most attempts to neutralize TrkB in tumors aim to inactivate its kinase activity, it is unclear whether TrkB kinase activity is required for its oncogenic functions. Indeed, it has been suggested that also other properties of the receptor contribute to functions that are relevant to tumor cell survival. Specifically, several adhesion motifs reside within the extracellular domains of TrkB. In line with this, TrkB-expressing epithelial cells form large cellular aggregates in suspension cultures, possibly facilitating tumor cell survival. Therefore, we set out to study the relative contributions of TrkBs kinase activity and its adhesion domains to anoikis suppression and oncogenicity. On the basis of a structure-function analysis, we report that TrkB kinase activity is required and, unexpectedly, also sufficient for anoikis suppression, tumor formation, and experimental metastasis. Thus, TrkB can act tumorigenically independent of its adhesion motifs. These results suggest that targeting the enzymatic activity of TrkB might be beneficial in cancer therapy.

PDGF suppresses the activation of group II phospholipase A2 gene expression by interleukin I and forskolin in mesangial cells

Treatment of rat mesangial cells with interleukin 1β (IL‐1β) and forskolin greatly enhanced the expression of group II phospholipase A2 (PLA2) mRNA, with subsequent increased synthesis and secretion of PLA2 as detected by PLA2 activity measurements and immunoprecipitation of culture media of [35S]methionine‐labelled mesangial cells. PDGF—BB dose‐dependently suppressed the IL‐1β‐ and forskolin‐induced elevation of PLA2 mRNA, as well as PLA2 synthesis and secretion. In contrast, PDGF—AA had no inhibitory effect. The tyrosine kinase inhibitor genistein dose‐dependently antagonized the inhibitory effect of PDGF‐BB on IL‐1β‐stimulated PLA2 secretion, thus suggesting that tyrosine phosphorylation may be required for PDGF‐BB inhibition of PLA2 gene expression in mesangial cells.

Transforming growth factors type-β and dexamethasone attenuate group II phospholipase A2 gene expression by interleukin-1 and forskolin in rat mesangial cells

Treatment of rat mesangial cells with interleukin‐1β(IL‐1β) and forskolin induced, in a synergistic fashion,the expression of group II phospholipase A2 (PLA2) mRNA, with subsequent increased synthesis and secretion of PLA2. In contrast, interleukin‐6 did not increase PLA2 mRNA levels of PLA2 activity. Transforming growth factor (TGF) β1, TGFβ2 and TGFβ3 equipotently attenuated the IL‐1β‐ and forskolin‐induced elevation of PLA2 mRNA, as well as PLA2 synthesis and secretion. The glucocorticoid dexamethasone only partially suppressed the IL‐1β‐ and forskolin‐induced elevation of PLA2 mRNA, but totally inhibited PLA2 synthesis and secretion.

Antisense- and antigene-based drug design strategies in oncology

Publisher Summary Cancer represents the second most frequent cause of death in the civilized countries and it is expected to become the leading mortal disease in the immediate future. The treatment of cancer presently is based on a combination of surgery, radiotherapy, and chemotherapy, with surgery and chemotherapy having been used in the treatment of localized, regional, and disseminated disease over the past 20 years with only minor innovative changes. The improved (though still highly insufficient) understanding of the factors contributing to carcinogenesis may now offer the possibility to design oligonucleotide-based therapeutic agents that could be largely devoid of the unspecific toxic side effects displayed by traditional anticancer agents. This chapter focuses on the use of exogenously delivered synthetic oligonucleotides as potential therapeutic agents. The chapter discusses related approaches involving—for example, the plasmid-derived endogenous expression of antisense RNA or the application of exogenously delivered large antisense RNA molecules.