Multiple roles for the receptor tyrosine kinase axl in tumor formation

A focus of contemporary cancer therapeutic development is the targeting of both the transformed cell and the supporting cellular microenvironment. Cell migration is a fundamental cellular behavior required for the complex interplay between multiple cell types necessary for tumor development. We therefore developed a novel retroviral-based screening technology in primary human endothelial cells to discover genes that control cell migration. We identified the receptor tyrosine kinase Axl as a novel regulator of endothelial cell haptotactic migration towards the matrix factor vitronectin. Using small interfering RNA-mediated silencing and overexpression of wild-type or mutated receptor proteins, we show that Axl is a key regulator of multiple angiogenic behaviors including endothelial cell migration, proliferation, and tube formation in vitro. Moreover, using sustained, retrovirally delivered short hairpin RNA (shRNA) Axl knockdown, we show that Axl is necessary for in vivo angiogenesis in a mouse model. Furthermore, we show that Axl is also required for human breast carcinoma cells to form a tumor in vivo. These findings indicate that Axl regulates processes vital for both neovascularization and tumorigenesis. Disruption of Axl signaling using a small-molecule inhibitor will hence simultaneously affect both the tumor and stromal cell compartments and thus represents a unique approach for cancer therapeutic development.     

Tibial subacute osteomyelitis with intraosseous abscess: an unusual complication of intraosseous infusion

Intravenous (IV) access is a critical step in patient care, especially in the emergency and/or trauma setting. Recently, intraosseous (IO) infusion has re-emerged as a recommended alternative to central venous access in both the pediatric and the adult patient. We present the case of an older adult male patient several months after emergency tibial IO infusion, now with left shin pain, and the MRI and culture findings diagnostic of subacute osteomyelitis with IO abscess, an unusual complication of IO infusion.     

Activation of the PKB/AKT pathway by ICAM-2

We identified intracellular adhesion molecule-2 (ICAM-2) in a genetic screen as an activator of the PI3K/AKT pathway leading to inhibition of apoptosis. ICAM-2 induced tyrosine phosphorylation of ezrin and PI3K kinase membrane translocation, resulting in phosphatidylinositol 3,4,5 production, PDK-1 and AKT activation, and subsequent phosphorylation of AKT targets BAD, GSK3, and FKHR. ICAM-2 clustering protected primary human CD19+ cells from TNFalpha- and Fas-mediated apoptosis as determined by single-cell analysis. ICAM-2 engagement by CD19+ cells of its natural receptor, LFA-1, on CD4+ naive cells specifically induced AKT activity in the absence of an MHC-peptide interaction. These results attribute a novel signaling function to ICAM-2 that might suggest mechanisms by which ICAM-2 signals intracellular communication at various immunological synapses.     

Postural destabilization induced by trunk extensor muscles fatigue is suppressed by use of a plantar pressure-based electro-tactile biofeedback

Separate studies have reported that postural control during quiet standing could be (1) impaired with muscle fatigue localized at the lower back, and (2) improved through the use of plantar pressure-based electro-tactile biofeedback, under normal neuromuscular state. The aim of this experiment was to investigate whether this biofeedback could reduce postural destabilization induced by trunk extensor muscles. Ten healthy adults were asked to stand as immobile as possible in four experimental conditions: (1) no fatigue/no biofeedback, (2) no fatigue/biofeedback, (3) fatigue/no biofeedback and (4) fatigue/biofeedback. Muscular fatigue was achieved by performing trunk repetitive extensions until maximal exhaustion. The underlying principle of the biofeedback consisted of providing supplementary information related to foot sole pressure distribution through electro-tactile stimulation of the tongue. Centre of foot pressure (CoP) displacements were recorded using a force platform. Results showed (1) increased CoP displacements along the antero-posterior axis in the fatigue than no fatigue condition in the absence of biofeedback and (2) no significant difference between the no fatigue and fatigue conditions in the presence of biofeedback. This suggests that subjects were able to efficiently integrate an artificial plantar pressure information delivered through electro-tactile stimulation of the tongue that allowed them to suppress the destabilizing effect induced by trunk extensor muscles fatigue.     

Mast cell tryptase regulates rat colonic myocytes through proteinase-activated receptor 2.

Proteinase-activated receptor-2 (PAR-2) is a G protein-coupled receptor that is cleaved and activated by trypsin-like enzymes. PAR-2 is highly expressed by small intestinal enterocytes where it is activated by luminal trypsin. The location, mechanism of activation, and biological functions of PAR-2 in the colon, however, are unknown. We localized PAR-2 to the muscularis externa of the rat colon by immunofluorescence. Myocytes in primary culture also expressed PAR-2, assessed by immunofluorescence and RT-PCR. Trypsin, SLIGRL-NH2 (corresponding to the PAR-2 tethered ligand), mast cell tryptase, and a filtrate of degranulated mast cells stimulated a prompt increase in [Ca2+]i in myocytes. The response to tryptase and the mast cell filtrate was inhibited by the tryptase inhibitor BABIM, and abolished by desensitization of PAR-2 with trypsin. PAR-2 activation inhibited the amplitude of rhythmic contractions of strips of rat colon. This response was unaffected by indomethacin, l-NG-nitroarginine methyl ester, a bradykinin B2 receptor antagonist and tetrodotoxin. Thus, PAR-2 is highly expressed by colonic myocytes where it may be cleaved and activated by mast cell tryptase. This may contribute to motility disturbances of the colon during conditions associated with mast cell degranulation.