Article
Mechanisms of Cytomegalovirus to tumour transmission across the blood-brain barrier in glioblastoma
Mechanismen des Cytomegalovirus zur Überwindung der Blut-Hirn-Schranke beim Glioblastom
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Authors
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Harald Krenzlin
- Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Neurochirurgische Klinik und Poliklinik, Mainz, Deutschland
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Leon Schmidt
- Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Neurochirurgische Klinik und Poliklinik, Mainz, Deutschland
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Thomas Berenz
- Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Neurochirurgische Klinik und Poliklinik, Mainz, Deutschland
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Niels Lemmermann
- Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Institut für Virologie, Mainz, Deutschland
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Sean Lawler
- Brigham and Women's Hospital, Department of Neurosurgery, Boston, MA, Vereinigte Staaten
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Florian Ringel
- Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Neurochirurgische Klinik und Poliklinik, Mainz, Deutschland
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Naureen Keric
- Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Neurochirurgische Klinik und Poliklinik, Mainz, Deutschland
| Published: | May 25, 2022 |
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Outline
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Objective: Cytomegalovirus (CMV) has been linked to glioblastoma for over a decade. However, mechanisms of how CMV tumor infection occurs are poorly understood. After infection, CMV persists in mononuclear myeloid and dendritic cell progenitors. Here we identify possible transmission pathways involving mononuclear cells and in transit infection of cells of the blood-brain barrier in human and murine in vitro and in vivo models.
Methods: In vivo, mononuclear cells from blood and bone marrow from CMV latent mice were harvested and transferred to CMV naïve animals carrying orthotopic Gl261fluc glioblastoma. Viral infection was analysed using immunofluorescence. In vitro, human cells of the BBB were co-culture forming spheroids mimicking the blood-brain barrier (3D spheroid BBB model). The viral transmission was analysed using fluorescence-labeled (mCherry and GFP) murine (MCMV) and human CMV (HCMV) viruses (GFP CMV) via live-cell imaging and cry section of murine brain slices.
Results: Bone marrow and peripheral blood mononuclear cells from CMV latent animals were positively stained for CMV using two immunofluorescence antibodies. Transfer of these cells into seronegative CMV naïve mice carrying Gl261 orthotopic tumors led to tumor infection and viral spreading within the glioblastoma. Incubation of BBB spheres consisting of human astrocytes, human brain vascular pericytes (hBVP), and human brain microvascular endothelial cells (hCMEC) with HCMV lead to sequential infection of these cells. In our model, endothelial cells on the outer perimeter were infected first, followed by hBVP, and then astrocytes located in the core of these spheres. Plaque assays and live-cell imaging showed lytic replication of HCMV in hBVP and hCMEC. Using Rhod-Dextran, we found the permeability of the blood-brain barrier reduced by 10.4% (p=0.0167) on day 3 and 39.23% (p=0.0046) on day seven after infection.
Conclusion: Mononuclear cells are a site of latent CMV infection from where infection can spread to glioblastomas. One mechanism of viral entry involves crossing the BBB and infection of perivascular cells in transit. THBVP and hCMEC might then maintain and drive the infection, while tightening the blood-brain barrier might interfere with chemotherapy.
