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Objective. The lack of an endometrial epithelium-specific promoter has slowed down the development of technically advanced mouse models of endometrial cancer. The aim of this study was to test whether direct in vivo adenoviral-mediated gene delivery can be used to circumvent this problem.Methods. Adenoviruses expressing the LacZ reporter gene or the Cre recombinase were injected into the left horn of the mouse uterus. Histochemistry and immunohistochemistry were used to detect expression of the reporter gene as well as targeted deletion of a floxed allele.Results. Our data demonstrate that in vivo direct injection of adenoviruses can efficiently target the endometrium in the mouse, specifically transducing genes to the glandular epithelial component.Conclusions. This approach will allow the generation of more refined and genetically defined mouse models of endometrial cancer. Endometrial gland-specific transient expression of recombinases, such as Cre, may thus be employed to delete engineered alleles of tumor suppressor genes and to activate the expression of latent oncogenes. [Copyright &y& Elsevier]

Genetically engineered and orthotopic xenograft mouse models have been instrumental for increasing our understanding of thyroid cancer progression and for the development of novel therapeutic approaches in a setting that is more physi ologically relevant than the classical subcutaneous flank implants. However, the anatomical location of the thyroid gland precludes a non-invasive analysis at the cellular level of the interactions between tumor cells and the surrounding microenvironment and does not allow a real-time evaluation of the response of tumor cells to drug treatments. A s a consequence, such studies have generally only relied on endpoint approaches, limiting the amount and depth of the information that could be gathered. Here we descri be the development of an innovative approach to imaging specific aspects of thyroid cancer biology, based on the implantation of a permanent, minimally invasive optical window that allows highresolution, multi-day, intravital imaging of the behavior and cellular dynamics of thyroid tumors in the mouse. We show that this technology allows visual ization of fluorescently tagged tumor cells both in immunocompetent, genetically engineered mouse models of anaplastic thyroid cancer (ATC) and in immunocompromised mice carrying orthotopic implanted human or mouse ATC cells. Furthermore, the use of rec ipient mice in which endothelial cells and macrophages are fluorescently labeled allows the detection of the spatial and functional relationship between tumor cells and their microenvironment. Finally, we show that ATC cells expressing a fluorescent biosensor for caspase 3 activity can be effectively utilized to evaluate, in real-time, the efficacy and kinetics of action of novel small molecule therapeutics. This novel approach to intravital imaging of thyroid cancer represents a platform that will allow, for the fi rst time, the longitudinal, in situ analysis of tumor cell responses to therapy and of their inter action with the microenvironment. [ABSTRACT FROM AUTHOR]

Anaplastic thyroid cancer (ATC) is among the most lethal malignancies. The mitotic kinase PLK1 is overexpressed in the majority of ATCs and PLK1 inhibitors have shown preclinical efficacy. However, they also cause mitotic slippage and endoreduplication, leading to the generation of tetraploid, genetically unstable cell populations. We hypothesized that PI3K activity may facilitate mitotic slippage upon PLK1 inhibition, and thus tested the effect of combining PLK1 and PI3K inhibitors in ATC models, in vitro and in vivo. Treatment with BI6727 and BKM120 resulted in a significant synergistic effect in ATC cells, independent of the levels of AKT activity. Combination of the two drugs enhanced growth suppression at doses for which the single drugs showed no effect, and led to a massive reduction of the tetraploid cells population. Furthermore, combined treatment in PI3Khigh cell lines showed a significant induction of apoptosis. Finally, combined inhibition of PI3K and PLK1 was extremely effective in vivo, in an immunocompetent allograft model of ATC. Our results demonstrate a clear therapeutic potential of combining PLK1 and PI3K inhibitors in anaplastic thyroid tumors. [ABSTRACT FROM AUTHOR]