Disruption of 3D tissue integrity facilitates adenovirus infection by deregulating the coxsackievirus and adenovirus receptor

The human coxsackievirus and adenovirus receptor (CAR) represents the primary cellular site of adenovirus attachment during infection. An understanding of the mechanisms regulating its expression could contribute to improving efficacy and safety of adenovirus-based therapies. We characterized regulation of CAR expression in a 3D cell culture model of human breast cancer progression, which mimics aspects of the physiological tissue context in vitro. Phenotypically normal breast epithelial cells (S1) and their malignant derivative (T4-2 cells) were grown either on tissue culture plastic (2D) or 3D cultures in basement membrane matrix. S1 cells grown in 3D showed low levels of CAR, which was expressed mainly at cell–cell junctions. In contrast, T4-2 cells expressed high levels of CAR, which was mainly in the cytoplasm. When signaling through the epidermal growth factor receptor was inhibited in T4-2 cells, cells reverted to a normal phenotype, CAR protein expression was significantly reduced, and the protein relocalized to cell–cell junctions. Growth of S1 cells as 2D cultures or in 3D in collagen-I, a nonphysiological microenvironment for these cells, led to up-regulation of CAR to levels similar to those in T4-2 cells, independently of cellular growth rates. Thus, expression of CAR depends on the integrity and polarity of the 3D organization of epithelial cells. Disruption of this organization by changes in the microenvironment, including malignant transformation, leads to up-regulation of CAR, thus enhancing the cell's susceptibility to adenovirus infection.

[1]  A. Fattaey,et al.  An Adenovirus Mutant That Replicates Selectively in p53- Deficient Human Tumor Cells , 1996, Science.

[2]  J. Bergelson,et al.  The dual impact of coxsackie and adenovirus receptor expression on human prostate cancer gene therapy. , 2000, Cancer research.

[3]  S. Hellman,et al.  Separating favorable from unfavorable prognostic markers in breast cancer: the role of E-cadherin. , 2000, Cancer research.

[4]  M. Bissell,et al.  Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[5]  G. Nemerow,et al.  Integrins α v β 3 and α v β 5 promote adenovirus internalization but not virus attachment , 1993, Cell.

[6]  A. Sagalowsky,et al.  The mechanism of the growth-inhibitory effect of coxsackie and adenovirus receptor (CAR) on human bladder cancer: a functional analysis of car protein structure. , 2001, Cancer research.

[7]  M. Mareel,et al.  Internalization of the E-Cadherin/Catenin Complex and Scattering of Human Mammary Carcinoma Cells MCF-7/AZ after Treatment with Conditioned Medium from Human Skin Squamous Carcinoma Cells COLO 16 , 2000, Cell adhesion and communication.

[8]  J. Bergelson,et al.  Isolation of a Common Receptor for Coxsackie B Viruses and Adenoviruses 2 and 5 , 1997, Science.

[9]  P. Bryant,et al.  Loss of the tight junction MAGUK ZO-1 in breast cancer: relationship to glandular differentiation and loss of heterozygosity. , 1998, The American journal of pathology.

[10]  O. Petersen,et al.  Trisomy 7p and malignant transformation of human breast epithelial cells following epidermal growth factor withdrawal. , 1996, Cancer research.

[11]  R. Kuwano,et al.  The coxsackievirus-adenovirus receptor protein as a cell adhesion molecule in the developing mouse brain. , 2000, Brain research. Molecular brain research.

[12]  C. Larabell,et al.  Reversion of the Malignant Phenotype of Human Breast Cells in Three-Dimensional Culture and In Vivo by Integrin Blocking Antibodies , 1997, The Journal of cell biology.

[13]  M J Bissell,et al.  Cellular growth and survival are mediated by beta 1 integrins in normal human breast epithelium but not in breast carcinoma. , 1995, Journal of cell science.

[14]  A. Ridley,et al.  Activation of both MAP kinase and phosphatidylinositide 3-kinase by Ras is required for hepatocyte growth factor/scatter factor-induced adherens junction disassembly. , 1998, Molecular biology of the cell.

[15]  D. R. Wilson,et al.  Tumor suppression and therapy sensitization of localized and metastatic breast cancer by adenovirus p53. , 2001, Human gene therapy.

[16]  N. Chapman,et al.  Expression of the Coxsackievirus and Adenovirus Receptor in Cultured Human Umbilical Vein Endothelial Cells: Regulation in Response to Cell Density , 1999, Journal of Virology.

[17]  D. Goodenough,et al.  Restoration of tight junction structure and barrier function by down-regulation of the mitogen-activated protein kinase pathway in ras-transformed Madin-Darby canine kidney cells. , 2000, Molecular biology of the cell.

[18]  Mina J Bissell,et al.  Normal and tumor-derived myoepithelial cells differ in their ability to interact with luminal breast epithelial cells for polarity and basement membrane deposition. , 2010, Journal of cell science.

[19]  M. Sachs,et al.  Integrin αv and coxsackie adenovirus receptor expression in clinical bladder cancer , 2002 .

[20]  N. Auersperg,et al.  E-cadherin induces mesenchymal-to-epithelial transition in human ovarian surface epithelium. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Gerhard Christofori,et al.  A causal role for E-cadherin in the transition from adenoma to carcinoma , 1998, Nature.

[22]  Deepika Walpita,et al.  Studying actin-dependent processes in tissue culture , 2002, Nature Reviews Molecular Cell Biology.

[23]  T. Godfrey,et al.  Quantitative mRNA expression analysis from formalin-fixed, paraffin-embedded tissues using 5' nuclease quantitative reverse transcription-polymerase chain reaction. , 2000, The Journal of molecular diagnostics : JMD.

[24]  R. Dummer,et al.  The presence of human coxsackievirus and adenovirus receptor is associated with efficient adenovirus-mediated transgene expression in human melanoma cell cultures. , 1998, Human gene therapy.

[25]  Kenneth M. Yamada,et al.  Taking Cell-Matrix Adhesions to the Third Dimension , 2001, Science.

[26]  Adenoviral GM-CSF gene transduction into breast cancer cells induced long-lasting antitumor immunity in mice , 1999, Breast cancer.

[27]  C. Larabell,et al.  Reciprocal interactions between beta1-integrin and epidermal growth factor receptor in three-dimensional basement membrane breast cultures: a different perspective in epithelial biology. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[28]  K. Chew,et al.  Expression of the coxsackie adenovirus receptor in normal prostate and in primary and metastatic prostate carcinoma: potential relevance to gene therapy. , 2002, Cancer research.

[29]  A. Sagalowsky,et al.  Loss of adenoviral receptor expression in human bladder cancer cells: a potential impact on the efficacy of gene therapy. , 1999, Cancer research.

[30]  J. Bergelson,et al.  The coxsackievirus and adenovirus receptor is a transmembrane component of the tight junction , 2001, Proceedings of the National Academy of Sciences of the United States of America.