Human motor neurons derived from induced pluripotent stem cells are susceptible to SARS-CoV-2 infection

Gioia Cappelletti; Claudia Colombrita; Fiona Limanaqi; Sabrina Invernizzi; Micaela Garziano; Claudia Vanetti; Claudia Moscheni; Serena Santangelo; Silvia Zecchini; Daria Trabattoni; Vincenzo Silani; Mario Clerici; Antonia Ratti; Mara Biasin

doi:10.3389/fncel.2023.1285836

Human motor neurons derived from induced pluripotent stem cells are susceptible to SARS-CoV-2 infection

Front Cell Neurosci. 2023 Dec 5:17:1285836. doi: 10.3389/fncel.2023.1285836. eCollection 2023.

Authors

Gioia Cappelletti¹, Claudia Colombrita², Fiona Limanaqi^{1

3}, Sabrina Invernizzi², Micaela Garziano^{1

3}, Claudia Vanetti¹, Claudia Moscheni¹, Serena Santangelo⁴, Silvia Zecchini¹, Daria Trabattoni¹, Vincenzo Silani^{2

5}, Mario Clerici^{3

6}, Antonia Ratti^{2

4}, Mara Biasin¹

Affiliations

¹ Laboratory of Immune-Biology, Department of Biomedical and Clinical Sciences, University of Milan, Milan, Italy.
² Department of Neurology and Laboratory of Neuroscience, IRCCS Istituto Auxologico Italiano, Milan, Italy.
³ Laboratory of Immunology, Department of Pathophysiology and Transplantation, University of Milan, Milan, Italy.
⁴ Department of Medical Biotechnology and Translational Medicine, Aldo Ravelli Center for Neurotechnology and Experimental Brain Therapeutics, University of Milan, Milan, Italy.
⁵ Department of Pathophysiology and Transplantation, "Dino Ferrari" Center, University of Milan, Milan, Italy.
⁶ Don C. Gnocchi Foundation, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Foundation, Milan, Italy.

Abstract

Introduction: COVID-19 typically causes Q7 respiratory disorders, but a high proportion of patients also reports neurological and neuromuscular symptoms during and after SARSCoV-2 infection. Despite a number of studies documenting SARS-CoV-2 infection of various neuronal cell populations, the impact of SARS-CoV-2 exposure on motor neuronal cells specifically has not been investigated so far.

Methods: Thus, by using human iPSC-derived motor neurons (iPSC-MNs) we assessed: (i) the expression of SARS-CoV-2 main receptors; (ii) iPSC-MN infectability by SARS-CoV-2; and (iii) the effect of SARS-CoV-2 exposure on iPSC-MN transcriptome.

Results: Gene expression profiling and immunofluorescence (IF) analysis of the main host cell receptors recognized by SARS-CoV-2 revealed that all of them are expressed in iPSC-MNs, with CD147 and NRP1 being the most represented ones. By analyzing SARS-CoV-2 N1 and N2 gene expression over time, we observed that human iPSC-MNs were productively infected by SARS-CoV-2 in the absence of cytopathic effect. Supernatants collected from SARS-CoV-2-infected iPSC-MNs were able to re-infect VeroE6 cells. Image analyses of SARS-CoV-2 nucleocapsid proteins by IF confirmed iPSC-MN infectability. Furthermore, SARS-CoV-2 infection in iPSCMNs significantly altered the expression of genes (IL-6, ANG, S1PR1, BCL2, BAX, Casp8, HLA-A, ERAP1, CD147, MX1) associated with cell survival and metabolism, as well as antiviral and inflammatory response.

Discussion: These results suggest for the very first time that SARS-CoV-2 can productively infect human iPSC-derived MNs probably by binding CD147 and NRP1 receptors. Such information will be important to unveil the biological bases of neuromuscular disorders characterizing SARS-CoV-2 infection and the so called long-COVID symptoms.

Keywords: COVID-19; SARS-CoV-2 infection; iPSC-derived motor neurons; long-COVID; neuroinflammation; neuromuscular disorders.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. Partially supported by grants from Fondazione Alessandro e Vincenzo Negroni Prati Morosini and Fondazione Romeo and Enrica Invernizzi, by EU funding within the Next Generation EU-MUR PNRR Extended Partnership initiative on Emerging Infectious Diseases (Project no. PE00000007, INF-ACT) and by the Italian Ministry of Health. SS was recipient of a fellowship from the Ph.D. program in Experimental Medicine of the University of Milan, Milan.