SARS-CoV-2 infects epithelial cells of the blood-cerebrospinal fluid barrier rather than endothelial cells or pericytes of the blood-brain barrier

Chiara Stüdle; Hideaki Nishihara; Sven Wischnewski; Laila Kulsvehagen; Sylvain Perriot; Hiroshi Ishikawa; Horst Schroten; Stephan Frank; Nikolaus Deigendesch; Renaud Du Pasquier; Lucas Schirmer; Anne-Katrin Pröbstel; Britta Engelhardt

doi:10.1186/s12987-023-00479-4

SARS-CoV-2 infects epithelial cells of the blood-cerebrospinal fluid barrier rather than endothelial cells or pericytes of the blood-brain barrier

Fluids Barriers CNS. 2023 Oct 24;20(1):76. doi: 10.1186/s12987-023-00479-4.

Authors

Chiara Stüdle¹, Hideaki Nishihara^{2

3}, Sven Wischnewski⁴, Laila Kulsvehagen⁵, Sylvain Perriot⁶, Hiroshi Ishikawa⁷, Horst Schroten⁸, Stephan Frank⁹, Nikolaus Deigendesch⁹, Renaud Du Pasquier^{6

10}, Lucas Schirmer^{4

11

12}, Anne-Katrin Pröbstel⁵, Britta Engelhardt¹³

Affiliations

¹ Theodor Kocher Institute, University of Bern, Bern, Switzerland. chiara.stuedle@unibe.ch.
² Theodor Kocher Institute, University of Bern, Bern, Switzerland.
³ Department of Neurotherapeutics, Yamaguchi University, Yamaguchi, Japan.
⁴ Department of Neurology, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
⁵ Departments of Neurology, Biomedicine and Clinical Research, Research Center for Clinical Neuroimmunology and Neuroscience Basel (RC2NB), University Hospital Basel and University of Basel, Basel, Switzerland.
⁶ Laboratory of Neuroimmunology, Neuroscience Research Centre, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV) and University of Lausanne, Lausanne, Switzerland.
⁷ Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, University of Tsukuba, Tsukuba, 305-8575, Ibaraki, Japan.
⁸ Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
⁹ Pathology, Institute of Medical Genetics and Pathology, University Hospital Basel and University of Basel, Basel, Switzerland.
¹⁰ Service of Neurology, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV), University of Lausanne, Lausanne, Switzerland.
¹¹ Center for Translational Neuroscience and Institute for Innate Immunoscience, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
¹² Interdisciplinary Center for Neurosciences, Heidelberg University, Heidelberg, Germany.
¹³ Theodor Kocher Institute, University of Bern, Bern, Switzerland. britta.engelhardt@unibe.ch.

Abstract

Background: As a consequence of SARS-CoV-2 infection various neurocognitive and neuropsychiatric symptoms can appear, which may persist for several months post infection. However, cell type-specific routes of brain infection and underlying mechanisms resulting in neuroglial dysfunction are not well understood.

Methods: Here, we investigated the susceptibility of cells constituting the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB) of the choroid plexus (ChP) to SARS-CoV-2 infection using human induced pluripotent stem cell (hiPSC)-derived cellular models and a ChP papilloma-derived epithelial cell line as well as ChP tissue from COVID-19 patients, respectively.

Results: We noted a differential infectibility of hiPSC-derived brain microvascular endothelial cells (BMECs) depending on the differentiation method. Extended endothelial culture method (EECM)-BMECs characterized by a complete set of endothelial markers, good barrier properties and a mature immune phenotype were refractory to SARS-CoV-2 infection and did not exhibit an activated phenotype after prolonged SARS-CoV-2 inoculation. In contrast, defined medium method (DMM)-BMECs, characterized by a mixed endothelial and epithelial phenotype and excellent barrier properties were productively infected by SARS-CoV-2 in an ACE2-dependent manner. hiPSC-derived brain pericyte-like cells (BPLCs) lacking ACE2 expression were not susceptible to SARS-CoV-2 infection. Furthermore, the human choroid plexus papilloma-derived epithelial cell line HIBCPP, modeling the BCSFB was productively infected by SARS-CoV-2 preferentially from the basolateral side, facing the blood compartment. Assessment of ChP tissue from COVID-19 patients by RNA in situ hybridization revealed SARS-CoV-2 transcripts in ChP epithelial and ChP stromal cells.

Conclusions: Our study shows that the BCSFB of the ChP rather than the BBB is susceptible to direct SARS-CoV-2 infection. Thus, neuropsychiatric symptoms because of COVID-19 may rather be associated with dysfunction of the BCSFB than the BBB. Future studies should consider a role of the ChP in underlying neuropsychiatric symptoms following SARS-CoV-2 infection.

Keywords: Blood-brain barrier; Blood-cerebrospinal fluid barrier; Choroid plexus epithelial cells; SARS-CoV-2; hiPSC-derived brain microvascular endothelial cells.

MeSH terms

Angiotensin-Converting Enzyme 2 / metabolism
Blood-Brain Barrier / metabolism
COVID-19*
Choroid Plexus / metabolism
Endothelial Cells / metabolism
Epithelial Cells / metabolism
Humans
Induced Pluripotent Stem Cells* / metabolism
Pericytes / metabolism
SARS-CoV-2 / metabolism

Substances

Angiotensin-Converting Enzyme 2

Grants and funding

ERC StG "DecOmPress/ERC_/European Research Council/International