RESUMO
Recent evidence has shown that many different tissues accumulate mutations even though the tissue is phenotypically normal. Therefore, generating mouse models for visualizing the tissue level effects that happen after oncogenic mutation in a single, isolated cell are critical for understanding tumor initiation and the role of competition in stem cell dynamics. Most mouse models have oncogenic mutations at the level of the entire mouse, the entire tissue, or all cells of a specific type in a tissue. However, these mouse models do not mimic the microenvironmental interactions that occur after an isolated cell acquires an oncogenic mutation because of the large number of mutant cells. We developed a mouse model for sporadic and isolated mutation of target alleles to better address the questions of sporadic cancer and stem cell competition. The following chapter describes methods for utilizing this mouse model and a few examples of the novel findings of using such a model.
Assuntos
Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Células-Tronco/metabolismo , Células-Tronco/patologia , Alelos , Animais , Animais Geneticamente Modificados , Modelos Animais de Doenças , Progressão da Doença , Neoplasias Intestinais/metabolismo , Neoplasias Intestinais/patologia , Intestino Delgado/metabolismo , Intestino Delgado/patologia , Camundongos , Mutação/genética , Mutação/fisiologiaRESUMO
Viral gene therapy is a means of delivering genes to replace malfunctioning ones, to kill cancer cells, or to correct genetic mutations. This technology is emerging as a powerful clinical tool; however, it is still limited by viral tropism, uptake and clearance by the liver, and most importantly an immune response. To overcome these challenges, we sought to merge the robustness of viral gene expression and the versatility of nanoparticle technology. Here, we describe a method for cloaking adenovirus (Ad) in silica (SiAd) as a nanoparticle formulation that significantly enhances transduction. Intratumoral injections in human glioma xenografts revealed SiAd expressing luciferase improved tumor transduction while reducing liver uptake. In immune-competent mice SiAd induced no inflammatory cytokines and reduced production of neutralizing antibodies. Finally, SiAd expressing TNF-related apoptosis-inducing ligand inhibited tumor growth of glioma xenografts. These results reveal that silica cloaking of Ad can enhance viral gene delivery while reducing immunogenicity.
