Inhibition of replication of hepatitis B virus in transgenic mice following administration of hepatotropic lipoplexes containing guanidinopropyl-modified siRNAs.
Chronic infection with hepatitis B virus (HBV) occurs commonly and complications that arise from persistence of the virus are associated with high mortality. Available licensed drugs have modest curative efficacy and advancing new therapeutic strategies to eliminate the virus is therefore a priority. HBV is susceptible to inactivation by exogenous gene silencers that harness RNA interference (RNAi) and the approach has therapeutic potential. To advance RNAi-based treatment for HBV infection, use in vivo of hepatotropic lipoplexes containing siRNAs with guanidinopropyl (GP) modifications is reported here. Lipoplexes contained polyglutamate, which has previously been shown to facilitate formulation and improve efficiency of the non-viral vectors. GP moieties were included in a previously described anti-HBV siRNA that effectively targeted the conserved viral X sequence. Particles had physical properties that were suitable for use in vivo: average diameter was approximately 50-200 nm and surface charge (zeta potential) was +65 mV. Efficient hepatotropic delivery of labeled siRNA was observed following systemic intravenous injection of the particles into HBV transgenic mice. Good inhibition of markers of viral replication was observed without evidence of toxicity. Efficacy of the GP-modified siRNAs was significantly more durable and formulations made up with chemically modified siRNAs were less immunostimulatory. An RNAi-mediated mechanism was confirmed by demonstrating that HBV mRNA cleavage occurred in vivo at the intended target site. Collectively these data indicate that GP-modified siRNAs formulated in anionic polymer-containing lipoplexes are effective silencers of HBV replication in vivo and have therapeutic potential.
Wits/SA MRC Antiviral Gene Therapy Research Unit, School of Pathology, Health Sciences Faculty, University of the Witwatersrand, Johannesburg, Private Bag 3, Wits 2050, South Africa.
Musa D
MD
Marimani
Wits/SA MRC Antiviral Gene Therapy Research Unit, School of Pathology, Health Sciences Faculty, University of the Witwatersrand, Johannesburg, Private Bag 3, Wits 2050, South Africa.
Abdullah
A
Ely
Goethe-University, Institute of Organic Chemistry & Chemical Biology, Max-von-Laue-Str. 7, 60438 Frankfurt am Main, Germany.
Maximilian C R
MC
Buff
Goethe-University, Institute of Organic Chemistry & Chemical Biology, Max-von-Laue-Str. 7, 60438 Frankfurt am Main, Germany.
Stefan
S
Bernhardt
Goethe-University, Institute of Organic Chemistry & Chemical Biology, Max-von-Laue-Str. 7, 60438 Frankfurt am Main, Germany.
Joachim W
JW
Engels
UTCBS, CNRS UMR8258, INSERM U1022, Université Paris Descartes, Chimie ParisTech, 75006 Paris, France.
Daniel
D
Scherman
UTCBS, CNRS UMR8258, INSERM U1022, Université Paris Descartes, Chimie ParisTech, 75006 Paris, France.
Virginie
V
Escriou
Wits/SA MRC Antiviral Gene Therapy Research Unit, School of Pathology, Health Sciences Faculty, University of the Witwatersrand, Johannesburg, Private Bag 3, Wits 2050, South Africa. Electronic address: Patrick.Arbuthnot@wits.ac.za.
Patrick
P
Arbuthnot
D000818:Animals; D016207:Cytokines / Q000378:metabolism; D006146:Guanidines / Q000737:chemistry*; D006514:Hepatitis B Surface Antigens / Q000097:blood; D006515:Hepatitis B virus / Q000502:physiology*; D008081:Liposomes; D008099:Liver / Q000378:metabolism; D008822:Mice, Transgenic; D010957:Plasmids; D011108:Polymers / Q000737:chemistry; D059367:RNA Cleavage; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D012367:RNA, Viral / Q000378:metabolism; D014779:Virus Replication
D016207:Cytokines; D006146:Guanidines; D006514:Hepatitis B Surface Antigens; D008081:Liposomes; D011108:Polymers; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral
Guanidinopropyl modification; HBV; Lipoplex; Polyglutamate; RNAi; SiRNA