Cytomegalovirus promotes murine glioblastoma growth via pericyte recruitment and angiogenesis
Harald Krenzlin, Prajna Behera, Viola Lorenz, Carmela Passaro, Mykola Zdioruk, Michal O. Nowicki, Korneel Grauwet, Hong Zhang, Magdalena Skubal, Hirotaka Ito, Rachel Zane, Michael Gutknecht, Marion B. Griessl, Franz Ricklefs, Lai Ding, Sharon Peled, Arun Rooj, C. David James, Charles S. Cobbs, Charles H. Cook, E. Antonio Chiocca, Sean E. Lawler
Harald Krenzlin, Prajna Behera, Viola Lorenz, Carmela Passaro, Mykola Zdioruk, Michal O. Nowicki, Korneel Grauwet, Hong Zhang, Magdalena Skubal, Hirotaka Ito, Rachel Zane, Michael Gutknecht, Marion B. Griessl, Franz Ricklefs, Lai Ding, Sharon Peled, Arun Rooj, C. David James, Charles S. Cobbs, Charles H. Cook, E. Antonio Chiocca, Sean E. Lawler
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Research Article Oncology Virology

Cytomegalovirus promotes murine glioblastoma growth via pericyte recruitment and angiogenesis

Abstract

Cytomegalovirus (CMV) has been implicated in glioblastoma (GBM); however, a mechanistic connection in vivo has not been established. The purpose of this study is to characterize the effects of murine CMV (MCMV) on GBM growth in murine models. Syngeneic GBM models were established in mice perinatally infected with MCMV. We found that tumor growth was markedly enhanced in MCMV+ mice, with a significant reduction in overall survival compared with that of controls (P < 0.001). We observed increased angiogenesis and tumor blood flow in MCMV+ mice. MCMV reactivation was observed in intratumoral perivascular pericytes and tumor cells in mouse and human GBM specimens, and pericyte coverage of tumor vasculature was strikingly augmented in MCMV+ mice. We identified PDGF-D as a CMV-induced factor essential for pericyte recruitment, angiogenesis, and tumor growth. The antiviral drug cidofovir improved survival in MCMV+ mice, inhibiting MCMV reactivation, PDGF-D expression, pericyte recruitment, and tumor angiogenesis. These data show that MCMV potentiates GBM growth in vivo by increased pericyte recruitment and angiogenesis due to alterations in the secretome of CMV-infected cells. Our model provides evidence for a role of CMV in GBM growth and supports the application of antiviral approaches for GBM therapy.

Authors

Harald Krenzlin, Prajna Behera, Viola Lorenz, Carmela Passaro, Mykola Zdioruk, Michal O. Nowicki, Korneel Grauwet, Hong Zhang, Magdalena Skubal, Hirotaka Ito, Rachel Zane, Michael Gutknecht, Marion B. Griessl, Franz Ricklefs, Lai Ding, Sharon Peled, Arun Rooj, C. David James, Charles S. Cobbs, Charles H. Cook, E. Antonio Chiocca, Sean E. Lawler

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Figure 2

MCMV infection accelerates GBM blood vessel formation in mice.

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MCMV infection accelerates GBM blood vessel formation in mice. (A) Ki67 ...
(A) Ki67 (green) immunofluorescence in brain sections taken from animals at the end point of survival studies. DAPI-stained nuclei are shown in blue. Scale bar: 50 μm. Graph shows Ki67-positive nuclei counts from 12 independent fields in 3 tumor samples. ***P < 0.005, Student’s t test. (B) CD31 (red) immunofluorescence in sections from GL261Luc2 tumors at survival end points (left panels). Scale bars: 100 μm. AngioTool analysis (right panels). Graphs show data for vascular parameters. Images are from 12 independent fields for each condition. n = 4. *P < 0.05; **P < 0.01 Student’s t test. (C) Kaplan-Meier survival curve of mice intracranially implanted with the murine CT-2A GBM cell line. Uninfected (n = 6) vs. MCMV+ (n = 6). P < 0.01, log-rank test. Median survival is indicated on plot and shown in parentheses. (D) Left panels show CD31 immunofluorescence staining (red) of tumor vasculature in 20 μm sections from CT-2A tumors in control and MCMV+ animals at survival end points. Scale bar: 100 μm. Graphs show representations of the data for vascular parameters based on analysis of angiogenesis and vessel morphology. Images are from 12 independent fields and 3 tumor specimens for each condition. *P < 0.05; ***P < 0.005, Student’s t test. (E) ASL-fMRI showing T2 coronal sections with ASL heatmap overlay. Quantitative analysis of both groups at corresponding ROIs. n = 3. Box extends from the 25th to 75th percentile, and the median is indicated by a horizontal line. Whiskers represent the maximum and minimum values. *P < 0.05, Student’s t test.