Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity

Juan R. Cubillos-Ruiz; Xavier Engle; Uciane K. Scarlett; Diana Martinez; Amorette Barber; Raul Elgueta; Li Wang; Yolanda Nesbeth; Yvon Durant; Andrew T. Gewirtz; Charles L. Sentman; Ross Kedl; Jose R. Conejo-Garcia

doi:10.1172/JCI37716

Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity

Juan R. Cubillos-Ruiz, Xavier Engle, Uciane K. Scarlett, Diana Martinez, Amorette Barber, Raul Elgueta, Li Wang, Yolanda Nesbeth, Yvon Durant, Andrew T. Gewirtz, Charles L. Sentman, Ross Kedl, Jose R. Conejo-Garcia

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Abstract

The success of clinically relevant immunotherapies requires reversing tumor-induced immunosuppression. Here we demonstrated that linear polyethylenimine-based (PEI-based) nanoparticles encapsulating siRNA were preferentially and avidly engulfed by regulatory DCs expressing CD11c and programmed cell death 1-ligand 1 (PD-L1) at ovarian cancer locations in mice. PEI-siRNA uptake transformed these DCs from immunosuppressive cells to efficient antigen-presenting cells that activated tumor-reactive lymphocytes and exerted direct tumoricidal activity, both in vivo and in situ. PEI triggered robust and selective TLR5 activation in vitro and elicited the production of hallmark TLR5-inducible cytokines in WT mice, but not in Tlr5-/- littermates. Thus, PEI is a TLR5 agonist that, to our knowledge, was not previously recognized. In addition, PEI-complexed nontargeting siRNA oligonucleotides stimulated TLR3 and TLR7. The nonspecific activation of multiple TLRs (specifically, TLR5 and TLR7) reversed the tolerogenic phenotype of human and mouse ovarian tumor-associated DCs. In ovarian carcinoma-bearing mice, this induced T cell-mediated tumor regression and prolonged survival in a manner dependent upon myeloid differentiation primary response gene 88 (MyD88; i.e., independent of TLR3). Furthermore, gene-specific siRNA-PEI nanocomplexes that silenced immunosuppressive molecules on mouse tumor-associated DCs elicited discernibly superior antitumor immunity and enhanced therapeutic effects compared with nontargeting siRNA-PEI nanocomplexes. Our results demonstrate that the intrinsic TLR5 and TLR7 stimulation of siRNA-PEI nanoparticles synergizes with the gene-specific silencing activity of siRNA to transform tumor-infiltrating regulatory DCs into DCs capable of promoting therapeutic antitumor immunity.

Original language	English
Pages (from-to)	2231-2244
Number of pages	14
Journal	The Journal of clinical investigation
Volume	119
Issue number	8
DOIs	https://doi.org/10.1172/JCI37716
Publication status	Published - 13 Aug 2009

Keywords

Animals
Antigen Presentation
Antigens, CD274
Antigens, CD80
Cytotoxicity, Immunologic
Dendritic Cells
Female
Humans
Membrane Glycoproteins
Mice
Mice, Inbred C57BL
Myeloid Differentiation Factor 88
Nanoparticles
Ovarian Neoplasms
Peptides
Polyethyleneimine
RNA, Small Interfering
T-Lymphocytes
Toll-Like Receptor 5

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1172/JCI37716

Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity Green PDFFinal published version, 4.56 MB

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Cubillos-Ruiz, J. R., Engle, X., Scarlett, U. K., Martinez, D., Barber, A., Elgueta, R., Wang, L., Nesbeth, Y., Durant, Y., Gewirtz, A. T., Sentman, C. L., Kedl, R., & Conejo-Garcia, J. R. (2009). Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity. The Journal of clinical investigation, 119(8), 2231-2244. https://doi.org/10.1172/JCI37716

@article{898814ec8e84417cb3936fbf410f20dd,

title = "Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity",

abstract = "The success of clinically relevant immunotherapies requires reversing tumor-induced immunosuppression. Here we demonstrated that linear polyethylenimine-based (PEI-based) nanoparticles encapsulating siRNA were preferentially and avidly engulfed by regulatory DCs expressing CD11c and programmed cell death 1-ligand 1 (PD-L1) at ovarian cancer locations in mice. PEI-siRNA uptake transformed these DCs from immunosuppressive cells to efficient antigen-presenting cells that activated tumor-reactive lymphocytes and exerted direct tumoricidal activity, both in vivo and in situ. PEI triggered robust and selective TLR5 activation in vitro and elicited the production of hallmark TLR5-inducible cytokines in WT mice, but not in Tlr5-/- littermates. Thus, PEI is a TLR5 agonist that, to our knowledge, was not previously recognized. In addition, PEI-complexed nontargeting siRNA oligonucleotides stimulated TLR3 and TLR7. The nonspecific activation of multiple TLRs (specifically, TLR5 and TLR7) reversed the tolerogenic phenotype of human and mouse ovarian tumor-associated DCs. In ovarian carcinoma-bearing mice, this induced T cell-mediated tumor regression and prolonged survival in a manner dependent upon myeloid differentiation primary response gene 88 (MyD88; i.e., independent of TLR3). Furthermore, gene-specific siRNA-PEI nanocomplexes that silenced immunosuppressive molecules on mouse tumor-associated DCs elicited discernibly superior antitumor immunity and enhanced therapeutic effects compared with nontargeting siRNA-PEI nanocomplexes. Our results demonstrate that the intrinsic TLR5 and TLR7 stimulation of siRNA-PEI nanoparticles synergizes with the gene-specific silencing activity of siRNA to transform tumor-infiltrating regulatory DCs into DCs capable of promoting therapeutic antitumor immunity.",

keywords = "Animals, Antigen Presentation, Antigens, CD274, Antigens, CD80, Cytotoxicity, Immunologic, Dendritic Cells, Female, Humans, Membrane Glycoproteins, Mice, Mice, Inbred C57BL, Myeloid Differentiation Factor 88, Nanoparticles, Ovarian Neoplasms, Peptides, Polyethyleneimine, RNA, Small Interfering, T-Lymphocytes, Toll-Like Receptor 5",

author = "Cubillos-Ruiz, {Juan R.} and Xavier Engle and Scarlett, {Uciane K.} and Diana Martinez and Amorette Barber and Raul Elgueta and Li Wang and Yolanda Nesbeth and Yvon Durant and Gewirtz, {Andrew T.} and Sentman, {Charles L.} and Ross Kedl and Conejo-Garcia, {Jose R.}",

year = "2009",

month = aug,

day = "13",

doi = "10.1172/JCI37716",

language = "English",

volume = "119",

pages = "2231--2244",

journal = "The Journal of clinical investigation",

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publisher = "The American Society for Clinical Investigation",

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Cubillos-Ruiz, JR, Engle, X, Scarlett, UK, Martinez, D, Barber, A, Elgueta, R, Wang, L, Nesbeth, Y, Durant, Y, Gewirtz, AT, Sentman, CL, Kedl, R & Conejo-Garcia, JR 2009, 'Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity', The Journal of clinical investigation, vol. 119, no. 8, pp. 2231-2244. https://doi.org/10.1172/JCI37716

TY - JOUR

T1 - Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity

AU - Cubillos-Ruiz, Juan R.

AU - Engle, Xavier

AU - Scarlett, Uciane K.

AU - Martinez, Diana

AU - Barber, Amorette

AU - Elgueta, Raul

AU - Wang, Li

AU - Nesbeth, Yolanda

AU - Durant, Yvon

AU - Gewirtz, Andrew T.

AU - Sentman, Charles L.

AU - Kedl, Ross

AU - Conejo-Garcia, Jose R.

PY - 2009/8/13

Y1 - 2009/8/13

N2 - The success of clinically relevant immunotherapies requires reversing tumor-induced immunosuppression. Here we demonstrated that linear polyethylenimine-based (PEI-based) nanoparticles encapsulating siRNA were preferentially and avidly engulfed by regulatory DCs expressing CD11c and programmed cell death 1-ligand 1 (PD-L1) at ovarian cancer locations in mice. PEI-siRNA uptake transformed these DCs from immunosuppressive cells to efficient antigen-presenting cells that activated tumor-reactive lymphocytes and exerted direct tumoricidal activity, both in vivo and in situ. PEI triggered robust and selective TLR5 activation in vitro and elicited the production of hallmark TLR5-inducible cytokines in WT mice, but not in Tlr5-/- littermates. Thus, PEI is a TLR5 agonist that, to our knowledge, was not previously recognized. In addition, PEI-complexed nontargeting siRNA oligonucleotides stimulated TLR3 and TLR7. The nonspecific activation of multiple TLRs (specifically, TLR5 and TLR7) reversed the tolerogenic phenotype of human and mouse ovarian tumor-associated DCs. In ovarian carcinoma-bearing mice, this induced T cell-mediated tumor regression and prolonged survival in a manner dependent upon myeloid differentiation primary response gene 88 (MyD88; i.e., independent of TLR3). Furthermore, gene-specific siRNA-PEI nanocomplexes that silenced immunosuppressive molecules on mouse tumor-associated DCs elicited discernibly superior antitumor immunity and enhanced therapeutic effects compared with nontargeting siRNA-PEI nanocomplexes. Our results demonstrate that the intrinsic TLR5 and TLR7 stimulation of siRNA-PEI nanoparticles synergizes with the gene-specific silencing activity of siRNA to transform tumor-infiltrating regulatory DCs into DCs capable of promoting therapeutic antitumor immunity.

AB - The success of clinically relevant immunotherapies requires reversing tumor-induced immunosuppression. Here we demonstrated that linear polyethylenimine-based (PEI-based) nanoparticles encapsulating siRNA were preferentially and avidly engulfed by regulatory DCs expressing CD11c and programmed cell death 1-ligand 1 (PD-L1) at ovarian cancer locations in mice. PEI-siRNA uptake transformed these DCs from immunosuppressive cells to efficient antigen-presenting cells that activated tumor-reactive lymphocytes and exerted direct tumoricidal activity, both in vivo and in situ. PEI triggered robust and selective TLR5 activation in vitro and elicited the production of hallmark TLR5-inducible cytokines in WT mice, but not in Tlr5-/- littermates. Thus, PEI is a TLR5 agonist that, to our knowledge, was not previously recognized. In addition, PEI-complexed nontargeting siRNA oligonucleotides stimulated TLR3 and TLR7. The nonspecific activation of multiple TLRs (specifically, TLR5 and TLR7) reversed the tolerogenic phenotype of human and mouse ovarian tumor-associated DCs. In ovarian carcinoma-bearing mice, this induced T cell-mediated tumor regression and prolonged survival in a manner dependent upon myeloid differentiation primary response gene 88 (MyD88; i.e., independent of TLR3). Furthermore, gene-specific siRNA-PEI nanocomplexes that silenced immunosuppressive molecules on mouse tumor-associated DCs elicited discernibly superior antitumor immunity and enhanced therapeutic effects compared with nontargeting siRNA-PEI nanocomplexes. Our results demonstrate that the intrinsic TLR5 and TLR7 stimulation of siRNA-PEI nanoparticles synergizes with the gene-specific silencing activity of siRNA to transform tumor-infiltrating regulatory DCs into DCs capable of promoting therapeutic antitumor immunity.

KW - Animals

KW - Antigen Presentation

KW - Antigens, CD274

KW - Antigens, CD80

KW - Cytotoxicity, Immunologic

KW - Dendritic Cells

KW - Female

KW - Humans

KW - Membrane Glycoproteins

KW - Mice

KW - Mice, Inbred C57BL

KW - Myeloid Differentiation Factor 88

KW - Nanoparticles

KW - Ovarian Neoplasms

KW - Peptides

KW - Polyethyleneimine

KW - RNA, Small Interfering

KW - T-Lymphocytes

KW - Toll-Like Receptor 5

U2 - 10.1172/JCI37716

DO - 10.1172/JCI37716

M3 - Article

C2 - 19620771

SN - 0021-9738

VL - 119

SP - 2231

EP - 2244

JO - The Journal of clinical investigation

JF - The Journal of clinical investigation

IS - 8

ER -

Polyethylenimine-based siRNA nanocomplexes reprogram tumor-associated dendritic cells via TLR5 to elicit therapeutic antitumor immunity

Abstract

Keywords

UN SDGs

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