TY - JOUR
T1 - Delivery of siRNAs to dendritic cells using DEC205-targeted lipid nanoparticles to inhibit immune responses
AU - Katakowski, Joseph A.
AU - Mukherjee, Gayatri
AU - Wilner, Samantha E.
AU - Maier, Keith E.
AU - Harrison, Michael Travis
AU - Di Lorenzo, Teresa P.
AU - Levy, Matthew
AU - Palliser, Deborah
N1 - Funding Information:
We thank Nahoko Dunlap (SRI International) for technical assistance with PBMC and moDC assays. We thank the Analytical Imaging Facility (AECOM) for generating cryo-EM images and the Flow Cytometry Facility (AECOM) for help with cell sorting. This work was supported in part by the NIH - R21 AI093539 and R01 AI099567 (D.P.); R21 CA157366, R21 CA157366 and Stand Up 2 Cancer (SU2C-AACRIRG0809) (M.L.); R01 DK094327, R01 DK064315, and R03 AI119225 (T.P.D.); P60 DK020541, which supports the Diabetes Research Center of the Albert Einstein College of Medicine; and P30 CA013330, which supports the flow cytometry facility; T32 NS007098 (J.A.K.); and by the Juvenile Diabetes Research Foundation (Academic R & D Award 17-2010-789 to T.P.D. and D.P.). T.P.D. is the Diane Belfer, Cypres and Endelson Families Faculty Scholar in Diabetes Research.
Funding Information:
We thank Nahoko Dunlap (SRI International) for technical assistance with PBMC and moDC assays. We thank the Analytical Imaging Facility (AECOM) for generating cryo-EM images and the Flow Cytometry Facility (AECOM) for help with cell sorting. This work was supported in part by the NIH - R21 AI093539 and R01 AI099567 (D.P.); R21 CA157366, R21 CA157366 and Stand Up 2 Cancer (SU2C-AACR-IRG0809) (M.L.); R01 DK094327, R01 DK064315, and R03 AI119225 (T.P.D.); P60 DK020541, which supports the Diabetes Research Center of the Albert Einstein College of Medicine; and P30 CA013330, which supports the flow cytometry facility; T32 NS007098 (J.A.K.); and by the Juvenile Diabetes Research Foundation (Academic R & D Award 17-2010-789 to T.P.D. and D.P.). T.P.D. is the Diane Belfer, Cypres and Endelson Families Faculty Scholar in Diabetes Research.
Publisher Copyright:
© 2016 The American Society of Gene & Cell Therapy.
PY - 2016/2/1
Y1 - 2016/2/1
N2 - Due to their ability to knock down the expression of any gene, siRNAs have been heralded as ideal candidates for treating a wide variety of diseases, including those involving "undruggable" targets. However, the therapeutic potential of siRNAs remains severely limited by a lack of effective delivery vehicles. Recently, lipid nanoparticles (LNPs) containing ionizable cationic lipids have been developed for hepatic siRNA delivery. However, their suitability for delivery to other cell types has not been determined. We have modified LNPs for preferential targeting to dendritic cells (DCs), central regulators of immune responses. To achieve directed delivery, we coated LNPs with a single-chain antibody (scFv; DEC-LNPs), specific to murine DEC205, which is highly expressed on distinct DC subsets. Here we show that injection of siRNAs encapsulated in DEC-LNPs are preferentially delivered to DEC205 + DCs. Gene knockdown following uptake of DEC-LNPs containing siRNAs specific for the costimulatory molecules CD40, CD80, and CD86 dramatically decreases gene expression levels. We demonstrate the functionality of this knockdown with a mixed lymphocyte response (MLR). Overall, we report that injection of LNPs modified to restrict their uptake to a distinct cell population can confer profound gene knockdown, sufficient to inhibit powerful immune responses like the MLR.
AB - Due to their ability to knock down the expression of any gene, siRNAs have been heralded as ideal candidates for treating a wide variety of diseases, including those involving "undruggable" targets. However, the therapeutic potential of siRNAs remains severely limited by a lack of effective delivery vehicles. Recently, lipid nanoparticles (LNPs) containing ionizable cationic lipids have been developed for hepatic siRNA delivery. However, their suitability for delivery to other cell types has not been determined. We have modified LNPs for preferential targeting to dendritic cells (DCs), central regulators of immune responses. To achieve directed delivery, we coated LNPs with a single-chain antibody (scFv; DEC-LNPs), specific to murine DEC205, which is highly expressed on distinct DC subsets. Here we show that injection of siRNAs encapsulated in DEC-LNPs are preferentially delivered to DEC205 + DCs. Gene knockdown following uptake of DEC-LNPs containing siRNAs specific for the costimulatory molecules CD40, CD80, and CD86 dramatically decreases gene expression levels. We demonstrate the functionality of this knockdown with a mixed lymphocyte response (MLR). Overall, we report that injection of LNPs modified to restrict their uptake to a distinct cell population can confer profound gene knockdown, sufficient to inhibit powerful immune responses like the MLR.
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U2 - 10.1038/mt.2015.175
DO - 10.1038/mt.2015.175
M3 - Article
C2 - 26412590
AN - SCOPUS:84957844235
SN - 1525-0016
VL - 24
SP - 146
EP - 155
JO - Molecular Therapy
JF - Molecular Therapy
IS - 1
ER -