TY - JOUR
T1 - Oncolytic H-1 parvovirus binds to sialic acid on laminins for cell attachment and entry
AU - Kulkarni, Amit
AU - Ferreira, Tiago
AU - Bretscher, Clemens
AU - Grewenig, Annabel
AU - El-Andaloussi, Nazim
AU - Bonifati, Serena
AU - Marttila, Tiina
AU - Palissot, Valérie
AU - Hossain, Jubayer A.
AU - Azuaje, Francisco
AU - Miletic, Hrvoje
AU - Ystaas, Lars A.R.
AU - Golebiewska, Anna
AU - Niclou, Simone P.
AU - Roeth, Ralf
AU - Niesler, Beate
AU - Weiss, Amélie
AU - Brino, Laurent
AU - Marchini, Antonio
N1 - Funding Information:
We thank Winfried Stocker (EuroImmun, Lübeck, Germany) for providing the pTriEX-1-LAMC1 plasmid, Richard Harbottle (DKFZ, Heidelberg, Germany) for the pCAG-S/ MAR cloning vector. We are also thankful to the team of the DKFZ Virus Production and Development Unit, in particular Marcus Müller, Barbara Liebetrau and Barbara Leuchs, for helping with virus production and titration and for providing the anti-H-1PV conformational antibody. We are grateful to Jean Rommelaere, Assia Angelova, Marcelo Ehrlich and Steeve Boulant for fruitful discussions. We also thank Sandra Caldeira and Caroline Hadley for critical reading of the manuscript. This study was supported initially by a seeding grant from Institut National du Cancer (INCA) to A.M. and L.B. and at later stages by a grant from ORYX GmbH to A.M. We would also like to express our deepest gratitude to André Welter for his generous donation.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/6/22
Y1 - 2021/6/22
N2 - H-1 parvovirus (H-1PV) is a promising anticancer therapy. However, in-depth understanding of its life cycle, including the host cell factors needed for infectivity and oncolysis, is lacking. This understanding may guide the rational design of combination strategies, aid development of more effective viruses, and help identify biomarkers of susceptibility to H-1PV treatment. To identify the host cell factors involved, we carry out siRNA library screening using a druggable genome library. We identify one crucial modulator of H-1PV infection: laminin γ1 (LAMC1). Using loss- and gain-of-function studies, competition experiments, and ELISA, we validate LAMC1 and laminin family members as being essential to H-1PV cell attachment and entry. H-1PV binding to laminins is dependent on their sialic acid moieties and is inhibited by heparin. We show that laminins are differentially expressed in various tumour entities, including glioblastoma. We confirm the expression pattern of laminin γ1 in glioblastoma biopsies by immunohistochemistry. We also provide evidence of a direct correlation between LAMC1 expression levels and H-1PV oncolytic activity in 59 cancer cell lines and in 3D organotypic spheroid cultures with different sensitivities to H-1PV infection. These results support the idea that tumours with elevated levels of γ1 containing laminins are more susceptible to H-1PV-based therapies.
AB - H-1 parvovirus (H-1PV) is a promising anticancer therapy. However, in-depth understanding of its life cycle, including the host cell factors needed for infectivity and oncolysis, is lacking. This understanding may guide the rational design of combination strategies, aid development of more effective viruses, and help identify biomarkers of susceptibility to H-1PV treatment. To identify the host cell factors involved, we carry out siRNA library screening using a druggable genome library. We identify one crucial modulator of H-1PV infection: laminin γ1 (LAMC1). Using loss- and gain-of-function studies, competition experiments, and ELISA, we validate LAMC1 and laminin family members as being essential to H-1PV cell attachment and entry. H-1PV binding to laminins is dependent on their sialic acid moieties and is inhibited by heparin. We show that laminins are differentially expressed in various tumour entities, including glioblastoma. We confirm the expression pattern of laminin γ1 in glioblastoma biopsies by immunohistochemistry. We also provide evidence of a direct correlation between LAMC1 expression levels and H-1PV oncolytic activity in 59 cancer cell lines and in 3D organotypic spheroid cultures with different sensitivities to H-1PV infection. These results support the idea that tumours with elevated levels of γ1 containing laminins are more susceptible to H-1PV-based therapies.
UR - http://www.scopus.com/inward/record.url?scp=85108334249&partnerID=8YFLogxK
UR - https://www.ncbi.nlm.nih.gov/pubmed/34158478
U2 - 10.1038/s41467-021-24034-7
DO - 10.1038/s41467-021-24034-7
M3 - Article
C2 - 34158478
AN - SCOPUS:85108334249
SN - 2041-1723
VL - 12
SP - 3834
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 3834
ER -