title: Synthesis and delivery of short, noncoding RNA by B lymphocytes.
abstract: Evolutionarily conserved short (20-30 nucleotides) noncoding RNAs (microRNAs) are powerful regulators of gene expression in a variety of physiological and pathological processes. As such, means to efficiently modulate microRNA function constitute an important therapeutic opportunity. Here we demonstrate that primary B lymphocytes can be genetically programmed with nonviral plasmid DNA for the biogenesis and delivery of antisense sequences (anti-microRNA) against microRNA-150 (miR-150). Within 18 h of transfection with an anti-miR-150 construct, primary B lymphocytes secrete ∼3,000 copies of anti-miR-150 molecules per cell. Anti-miR-150 molecules released by B lymphocytes were internalized by CD8 T lymphocytes during cross-priming in vitro and in vivo, resulting in marked down-regulation of endogenous miR-150. However, such internalization was not observed in the absence of cross-priming. These results suggest that shuttling anti-miR-150 molecules from B lymphocytes to T cells requires the activation of receiver T cells via the antigen receptor. Finally, anti-miR-150 synthesized in B cells were secreted both as free and extracellular vesicle-associated fractions, but only extracellular vesicle-associated anti-miR-150 were apparently taken up by CD8 T cells. Collectively, these data indicate that primary B lymphocytes represent an efficient platform for the synthesis and delivery of short, noncoding RNA, paving the way for an approach to immunogenomic therapies.
authors.affiliation: Laboratory of Immunology, Department of Medicine and Moores Cancer Center, and Center for Theoretical and Applied Neuro-Oncology, Division of Neurosurgery and Moores Cancer Center, University of California, San Diego, La Jolla, CA 92093-0815.
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Almanza
authors.forename: Veronika
authors.initials: V
authors.lastname: Anufreichik
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Rodvold
authors.forename: Kevin T
authors.initials: KT
authors.lastname: Chiu
authors.forename: Alexandra
authors.initials: A
authors.lastname: DeLaney
authors.forename: Johnny C
authors.initials: JC
authors.lastname: Akers
authors.forename: Clark C
authors.initials: CC
authors.lastname: Chen
authors.forename: Maurizio
authors.initials: M
authors.lastname: Zanetti
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001402:B-Lymphocytes / Q000378:metabolism*; D045142:Cross-Priming; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009841:Oligonucleotides / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D058727:RNA, Small Untranslated / Q000096:biosynthesis* / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection
chemical_list: D000906:Antibodies; D035683:MicroRNAs; D009841:Oligonucleotides; D058727:RNA, Small Untranslated; C533097:MIR150, human
keywords: immunotherapy; microvesicles