modeling-astrocytoma-pathogenesis-vitro-vivo-using-cortical

Modeling Astrocytoma Pathogenesis <em>In Vitro</em> and <em>In Vivo</em> Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

Current astrocytoma models are limited in their ability to define the roles of oncogenic mutations in specific brain cell types during disease ...

University of North Carolina School of Medicine

Immune surveillance of the airways is critical to maintain the integrity and health of the lung. We have identified a family of ligands expressed on the surface of stressed airway epithelial cells whose function is to bind the NKG2D-activating receptor found on several pulmonary lymphocytes, including natural killer cells, gammadelta(+) T cells, and CD8(+) T cells. We employed real-time PCR and flow cytometry in normal and transformed airway epithelial cell to demonstrate that major histocompatibility complex class I chain-related (MIC) B and the UL-16 binding protein (ULBP) ligands (ULBP1-4) are ubiquitously expressed at the mRNA level in all cell lines. MICA/B surface expression was present on 70% of transformed cell lines but was undetectable on primary cells. We demonstrate that MICA/B and ULBP 1, 2, 3, and 4 expression is rare or absent on the cell surface of unstimulated normal human bronchial epithelial cells although transcripts and intracellular proteins are present. Normal human bronchial epithelial cells exposed to 0.3 mM hydrogen peroxide exhibit an induction of all ligands examined on the cell surface. Surface expression is independent of changes in transcript level or total cellular protein and is mediated by the ERK family of mitogen-activated protein kinases. The induction of NKG2D ligands on stressed airway epithelial cells represents a potentially important mechanism of immune cell activation in regulation of pulmonary health and disease.

NKG2D ligands are expressed on stressed human airway epithelial cells.

Persistent macrophage accumulation and alveolar enlargement are hallmark features of chronic obstructive pulmonary disease (COPD). A role for CD8(+) lymphocytes in the development of COPD is suggested based on observations that this T cell subset is increased in the airways and parenchyma of smokers that develop COPD with airflow limitation. In this study, we utilize a mouse model of COPD to examine the contributions of CD8(+) T cells in the persistent macrophage accumulation and airspace enlargement resulting from chronic irritant exposure.

CD8+ T cells contribute to macrophage accumulation and airspace enlargement following repeated irritant exposure.

Chronic obstructive pulmonary disease (COPD) is a lethal progressive lung disease culminating in permanent airway obstruction and alveolar enlargement. Previous studies suggest CTL involvement in COPD progression; however, their precise role remains unknown. Here, we investigated whether the CTL activation receptor NK cell group 2D (NKG2D) contributes to the development of COPD. Using primary murine lung epithelium isolated from mice chronically exposed to cigarette smoke and cultured epithelial cells exposed to cigarette smoke extract in vitro, we demonstrated induced expression of the NKG2D ligand retinoic acid early transcript 1 (RAET1) as well as NKG2D-mediated cytotoxicity. Furthermore, a genetic model of inducible RAET1 expression on mouse pulmonary epithelial cells yielded a severe emphysematous phenotype characterized by epithelial apoptosis and increased CTL activation, which was reversed by blocking NKG2D activation. We also assessed whether NKG2D ligand expression corresponded with pulmonary disease in human patients by staining airway and peripheral lung tissues from never smokers, smokers with normal lung function, and current and former smokers with COPD. NKG2D ligand expression was independent of NKG2D receptor expression in COPD patients, demonstrating that ligand expression is the limiting factor in CTL activation. These results demonstrate that aberrant, persistent NKG2D ligand expression in the pulmonary epithelium contributes to the development of COPD pathologies.

Sustained CTL activation by murine pulmonary epithelial cells promotes the development of COPD-like disease.

Over the last decade, genetically engineered mouse models have been extensively used to dissect the genetic requirements for neoplastic initiation and progression of diffuse gliomas. While these models faithfully recapitulate the histopathological features of human gliomas, comparative genomic analyses are increasingly being utilized to comprehensively assess their fidelity to recently identified molecular subtypes of these tumors. Future progress with these models will rely on incorporating insights not only from oncogenomics studies of cancer, but also from the developmental neuroscience and stem cell biology fields to design accurate and experimentally tractable models for use in translational cancer research, particularly for experimental therapeutics studies of molecularly defined subtypes of gliomas.

Genetically engineered mouse models of diffuse gliomas.

Glioblastoma (GBM) genomes feature recurrent genetic alterations that dysregulate core intracellular signaling pathways, including the G1/S cell cycle checkpoint and the MAPK and PI3K effector arms of receptor tyrosine kinase (RTK) signaling. Elucidation of the phenotypic consequences of activated RTK effectors is required for the design of effective therapeutic and diagnostic strategies.

Article	Year
NKG2D ligands are expressed on stressed human airway epithelial cells. American journal of physiology. Lung cellular and molecular physiology\| PubMed ID: 16473864	2006
CD8+ T cells contribute to macrophage accumulation and airspace enlargement following repeated irritant exposure. Experimental and molecular pathology\| PubMed ID: 17950725	2007
Sustained CTL activation by murine pulmonary epithelial cells promotes the development of COPD-like disease. The Journal of clinical investigation\| PubMed ID: 19197141	2009
Genetically engineered mouse models of diffuse gliomas. Brain research bulletin\| PubMed ID: 21684324	2012
Cooperativity between MAPK and PI3K signaling activation is required for glioblastoma pathogenesis. Neuro-oncology\| PubMed ID: 23814263	2013

Mark Vitucci

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