Systemic administration of combinatorial dsiRNAs via nanoparticles efficiently suppresses HIV-1 infection in humanized mice.

We evaluated the in vivo efficacy of structurally flexible, cationic PAMAM dendrimers as a small interfering RNA (siRNA) delivery system in a Rag2(-)/-γc-/- (RAG-hu) humanized mouse model for HIV-1 infection. HIV-infected humanized Rag2-/-γc-/- mice (RAG-hu) were injected intravenously (i.v.) with dendrimer-siRNA nanoparticles consisting of a cocktail of dicer substrate siRNAs (dsiRNAs) targeting both viral and cellular transcripts. We report in this study that the dendrimer-dsiRNA treatment suppressed HIV-1 infection by several orders of magnitude and protected against viral induced CD4(+) T-cell depletion. We also demonstrated that follow-up injections of the dendrimer-cocktailed dsiRNAs following viral rebound resulted in complete inhibition of HIV-1 titers. Biodistribution studies demonstrate that the dendrimer-dsiRNAs preferentially accumulate in peripheral blood mononuclear cells (PBMCs) and liver and do not exhibit any discernable toxicity. These data demonstrate for the first time efficacious combinatorial delivery of anti-host and -viral siRNAs for HIV-1 treatment in vivo. The dendrimer delivery approach therefore represents a promising method for systemic delivery of combinations of siRNAs for treatment of HIV-1 infection.

[1]  Dale L. Greiner,et al.  T Cell-Specific siRNA Delivery Suppresses HIV-1 Infection in Humanized Mice , 2008, Cell.

[2]  J. Garcia,et al.  Novel humanized murine models for HIV research , 2009, Current HIV/AIDS reports.

[3]  A. Judge,et al.  siRNA and innate immunity. , 2009, Oligonucleotides.

[4]  Judy Lieberman,et al.  Interfering with disease: a progress report on siRNA-based therapeutics , 2007, Nature Reviews Drug Discovery.

[5]  A. Fire,et al.  Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans , 1998, Nature.

[6]  M. Martínez.  Progress in the Therapeutic Applications of siRNAs Against HIV-1 , 2008, Methods in molecular biology.

[7]  Ling Peng,et al.  PAMAM dendrimers for efficient siRNA delivery and potent gene silencing. , 2006, Chemical communications.

[8]  V. Hornung,et al.  siRNA and isRNA: two edges of one sword. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[9]  J. Lieberman,et al.  Identification of Host Proteins Required for HIV Infection Through a Functional Genomic Screen , 2007, Science.

[10]  Zeger Debyser,et al.  Transportin-SR2 Imports HIV into the Nucleus , 2008, Current Biology.

[11]  Cheng-Cai Zhang,et al.  Importance of size-to-charge ratio in construction of stable and uniform nanoscale RNA/dendrimer complexes. , 2007, Organic & biomolecular chemistry.

[12]  Ling Peng,et al.  PAMAM Dendrimers Mediate siRNA Delivery to Target Hsp27 and Produce Potent Antiproliferative Effects on Prostate Cancer Cells , 2009, ChemMedChem.

[13]  J. Rossi,et al.  An Aptamer-siRNA Chimera Suppresses HIV-1 Viral Loads and Protects from Helper CD4+ T Cell Decline in Humanized Mice , 2011, Science Translational Medicine.

[14]  P. Heegaard,et al.  Dendrimers in drug research. , 2004, Chemical Society reviews.

[15]  Ben Berkhout,et al.  Human Immunodeficiency Virus Type 1 Escapes from RNA Interference-Mediated Inhibition , 2004, Journal of Virology.

[16]  P. Ortega,et al.  Characterization of carbosilane dendrimers as effective carriers of siRNA to HIV-infected lymphocytes. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[17]  John J. Rossi,et al.  Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells , 2009, Nucleic acids research.

[18]  B. Palmer,et al.  HIV-1 infection and CD4 T cell depletion in the humanized Rag2-/-γc-/- (RAG-hu) mouse model , 2006, Retrovirology.

[19]  S. Mehandru,et al.  Role of the gastrointestinal tract in establishing infection in primates and humans , 2008, Current opinion in HIV and AIDS.

[20]  Ling Peng,et al.  Polycationic dendrimers interact with RNA molecules: polyamine dendrimers inhibit the catalytic activity of Candida ribozymes. , 2005, Chemical communications.

[21]  J. Lieberman,et al.  siRNA-directed inhibition of HIV-1 infection , 2002, Nature Medicine.

[22]  David P. Bartel,et al.  Passenger-Strand Cleavage Facilitates Assembly of siRNA into Ago2-Containing RNAi Enzyme Complexes , 2005, Cell.

[23]  John J. Rossi,et al.  Strategies for silencing human disease using RNA interference , 2007, Nature Reviews Genetics.

[24]  A. Eguchi,et al.  Efficient siRNA Delivery into Primary Cells by Peptide Transduction-dsRNA Binding Domain (PTD-DRBD) Fusion Protein , 2009, Nature Biotechnology.

[25]  Daniel G. Anderson,et al.  Knocking down barriers: advances in siRNA delivery , 2009, Nature Reviews Drug Discovery.

[26]  K. Kehn-Hall,et al.  The utilization of humanized mouse models for the study of human retroviral infections , 2009, Retrovirology.

[27]  J. Rossi.  RNAi as a treatment for HIV-1 infection. , 2006, BioTechniques.

[28]  G. Rettig,et al.  Progress Toward In Vivo Use of siRNAs-II , 2006, Molecular Therapy.

[29]  C. Apetrei,et al.  Mucosal immune dysfunction in AIDS pathogenesis. , 2008, AIDS reviews.

[30]  M. Manns,et al.  Humanized mice for modeling human infectious disease: challenges, progress, and outlook. , 2009, Cell host & microbe.

[31]  John J. Rossi,et al.  The promises and pitfalls of RNA-interference-based therapeutics , 2009, Nature.

[32]  D. Peer,et al.  RNAi-mediated CCR5 silencing by LFA-1-targeted nanoparticles prevents HIV infection in BLT mice. , 2010, Molecular therapy : the journal of the American Society of Gene Therapy.

[33]  J. Rossi,et al.  Progress and prospects: RNA-based therapies for treatment of HIV infection , 2007, Gene Therapy.

[34]  Sangdun Choi,et al.  Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy , 2005, Nature Biotechnology.

[35]  Hongjie Dai,et al.  siRNA delivery into human T cells and primary cells with carbon-nanotube transporters. , 2007, Angewandte Chemie.

[36]  E. Connick,et al.  Mucosal transmission of R5 and X4 tropic HIV-1 via vaginal and rectal routes in humanized Rag2-/- gammac -/- (RAG-hu) mice. , 2008, Virology.

[37]  T. Tuschl,et al.  Human Argonaute2 mediates RNA cleavage targeted by miRNAs and siRNAs. , 2004, Molecular cell.

[38]  John J Rossi,et al.  Novel dual inhibitory function aptamer-siRNA delivery system for HIV-1 therapy. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[39]  P. Sharp,et al.  RNAi Double-Stranded RNA Directs the ATP-Dependent Cleavage of mRNA at 21 to 23 Nucleotide Intervals , 2000, Cell.

[40]  Judy Lieberman,et al.  Antibody mediated in vivo delivery of small interfering RNAs via cell-surface receptors , 2005, Nature Biotechnology.

[41]  B. Berkhout,et al.  Towards a durable RNAi gene therapy for HIV-AIDS , 2009, Expert opinion on biological therapy.

[42]  M. Morilla,et al.  Ethylendiamine core PAMAM dendrimers/siRNA complexes as in vitro silencing agents. , 2009, International journal of pharmaceutics.

[43]  John J Rossi,et al.  Control of HIV-1 replication by RNA interference. , 2004, Virus research.

[44]  John J Rossi,et al.  Genetic therapies against HIV , 2007, Nature Biotechnology.

[45]  A. Banerjea,et al.  Bispecific short hairpin siRNA constructs targeted to CD4, CXCR4, and CCR5 confer HIV-1 resistance. , 2003, Oligonucleotides.