Targeted inhibition of hantavirus replication and intracranial pathogenesis by a chimeric protein-delivered siRNA.

Abstract

Hantavirus (HV) infection, which underlies hantavirus hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome, remains to be a severe clinical challenge. Here, we synthesized small interfering RNAs (siRNAs) that target the encoding sequences of HV strain 76-118, and validated their inhibitory role in virus replication in HV-infected monkey kidney Vero E6 cells. A chimeric protein, 3G1-Cκ-tP, consisting of a single-chain antibody fragment (3G1) against the HV surface envelop glycoprotein, the constant region of human immunoglobulin κ chain (Cκ), and truncated protamine (amino acids 8-29, tP), was further generated. The fusion protein showed high affinity to HV antigen on the infected cell membrane, and internalized through clathrin-mediated endocytosis; it bound to siRNAs via the basic nucleic acid-rich protamine fragment, leading to their specific delivery into HV-infected cells and efficient inhibition of virus replication. An encephalitis mouse model was established via intracranial HV administration. Intraperitoneal injection of siRNAs complexed with 3G1-Cκ-tP achieved specific distribution of siRNAs in HV-infected brain cells, significantly reduced HV antigen levels, and effective protection from HV infection-derived animal death. These results provide a compelling rationale for novel therapeutic protocols designed for HV infection and related disorders.

• Authors: Jie Yang, Ji-Feng Sun, Ting-Ting Wang, Xiao-Hong Guo, Jun-Xia Wei, Lin-Tao Jia, An-Gang Yang
• Journal: Antiviral research (Antiviral Res) Vol. 147 Pg. 107-115 (Nov 2017) ISSN: 1872-9096 [Electronic] Netherlands
• PMID: 29017779 (Publication Type: Journal Article)
• Copyright: Copyright © 2017 Elsevier B.V. All rights reserved.

Chemical References

• Antigens, Viral
• RNA, Small Interfering
• Recombinant Proteins
• Single-Chain Antibodies
• Viral Envelope Proteins

Topics

• Animals
• Animals, Newborn
• Antigens, Viral (metabolism)
• Chlorocebus aethiops
• Disease Models, Animal
• Drug Delivery Systems
• Orthohantavirus (drug effects)
• Hemorrhagic Fever with Renal Syndrome (drug therapy, pathology, virology)
• Humans
• Mice
• Mice, Inbred BALB C
• Protein Binding
• RNA Interference
• RNA, Small Interfering (administration & dosage, chemistry, genetics, pharmacology)
• Recombinant Proteins (administration & dosage, genetics, pharmacology)
• Single-Chain Antibodies (genetics)
• Vero Cells
• Viral Envelope Proteins (genetics)
• Viral Load (drug effects)
• Virus Replication (drug effects)