Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target

Research output: Contribution to journalArticle

Nuphar Veiga, Meir Goldsmith, Yael Diesendruck, Srinivas Ramishetti, Daniel Rosenblum, Eran Elinav, Mark A. Behlke, Itai Benhar, Dan Peer

Interferon regulatory factor 8 (IRF8) protein plays a critical role in the differentiation, polarization, and activation of mononuclear phagocytic cells. In light of previous studies, we explored the therapeutic potential of IRF8 inhibition as immunomodulatory therapy for inflammatory bowel disease (IBD). To this end, we utilized siRNA-loaded lipid-based nanoparticles (siLNPs) and demonstrated a ∼90% reduction of IRF8 mRNA levels in vitro (PV < 0.0001), alongside a notable reduction in IRF8 protein. Moreover, silencing IRF8 ex vivo in splenocytes lead to a profound downregulation of IRF8 protein, followed by an immunomodulatory effect, as represented by a decrease in the secretion of TNFα, IL6 and IL12/IL23 (IL12p40) proinflammatory cytokines (PV = 0.0045, 0.0330, <0.0001, respectively). In order to silence IRF8 in vivo, selectively in inflammatory leukocytes, we used siLNPs that were coated with anti-Ly6C antibodies via our recently published ASSET targeting approach. Through this strategy, we have demonstrated a selective binding of the targeted-LNPs (T-LNPs) to Ly6C + inflammatory leukocytes. Finally, an immunomodulatory effect was demonstrated in vivo in an IBD mouse model with a profound decrease of TNFα, IL6, IL12/IL23, and IL1β pro-inflammatory cytokines (n = 5, PV < 0.0001, <0.0001, <0.0001, 0.02, respectively) and an improvement of colon-morphology as assessed by colon-length measurements and colonoscopy (PV < 0.0001). Overall, using antibody-targeted siLNPs, we showed a notable reduction of IRF8 mRNA and protein and demonstrated a targeted immunomodulation therapeutic effect ex vivo and in vivo, in the DSS colitis model. We claim that a selective silencing of IRF8 in inflammatory leukocytes (such as Ly6C+) may serve as a therapeutic approach for treating inflammatory disorders.

Original languageEnglish (US)
Pages (from-to)33-41
Number of pages9
JournalJournal of Controlled Release
Volume313
DOIs
StatePublished - Nov 10 2019

PMID: 31634546

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Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target. / Veiga, Nuphar; Goldsmith, Meir; Diesendruck, Yael; Ramishetti, Srinivas; Rosenblum, Daniel; Elinav, Eran; Behlke, Mark A.; Benhar, Itai; Peer, Dan.

In: Journal of Controlled Release, Vol. 313, 10.11.2019, p. 33-41.

Research output: Contribution to journalArticle

Harvard

Veiga, N, Goldsmith, M, Diesendruck, Y, Ramishetti, S, Rosenblum, D, Elinav, E, Behlke, MA, Benhar, I & Peer, D 2019, 'Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target' Journal of Controlled Release, vol. 313, pp. 33-41. https://doi.org/10.1016/j.jconrel.2019.10.001

APA

Veiga, N., Goldsmith, M., Diesendruck, Y., Ramishetti, S., Rosenblum, D., Elinav, E., ... Peer, D. (2019). Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target. Journal of Controlled Release, 313, 33-41. https://doi.org/10.1016/j.jconrel.2019.10.001

Vancouver

Veiga N, Goldsmith M, Diesendruck Y, Ramishetti S, Rosenblum D, Elinav E et al. Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target. Journal of Controlled Release. 2019 Nov 10;313:33-41. https://doi.org/10.1016/j.jconrel.2019.10.001

Author

Veiga, Nuphar ; Goldsmith, Meir ; Diesendruck, Yael ; Ramishetti, Srinivas ; Rosenblum, Daniel ; Elinav, Eran ; Behlke, Mark A. ; Benhar, Itai ; Peer, Dan. / Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target. In: Journal of Controlled Release. 2019 ; Vol. 313. pp. 33-41.

BibTeX

@article{a3c173a73c5947eab2c35a0af60074aa,
title = "Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target",
abstract = "Interferon regulatory factor 8 (IRF8) protein plays a critical role in the differentiation, polarization, and activation of mononuclear phagocytic cells. In light of previous studies, we explored the therapeutic potential of IRF8 inhibition as immunomodulatory therapy for inflammatory bowel disease (IBD). To this end, we utilized siRNA-loaded lipid-based nanoparticles (siLNPs) and demonstrated a ∼90{\%} reduction of IRF8 mRNA levels in vitro (PV < 0.0001), alongside a notable reduction in IRF8 protein. Moreover, silencing IRF8 ex vivo in splenocytes lead to a profound downregulation of IRF8 protein, followed by an immunomodulatory effect, as represented by a decrease in the secretion of TNFα, IL6 and IL12/IL23 (IL12p40) proinflammatory cytokines (PV = 0.0045, 0.0330, <0.0001, respectively). In order to silence IRF8 in vivo, selectively in inflammatory leukocytes, we used siLNPs that were coated with anti-Ly6C antibodies via our recently published ASSET targeting approach. Through this strategy, we have demonstrated a selective binding of the targeted-LNPs (T-LNPs) to Ly6C + inflammatory leukocytes. Finally, an immunomodulatory effect was demonstrated in vivo in an IBD mouse model with a profound decrease of TNFα, IL6, IL12/IL23, and IL1β pro-inflammatory cytokines (n = 5, PV < 0.0001, <0.0001, <0.0001, 0.02, respectively) and an improvement of colon-morphology as assessed by colon-length measurements and colonoscopy (PV < 0.0001). Overall, using antibody-targeted siLNPs, we showed a notable reduction of IRF8 mRNA and protein and demonstrated a targeted immunomodulation therapeutic effect ex vivo and in vivo, in the DSS colitis model. We claim that a selective silencing of IRF8 in inflammatory leukocytes (such as Ly6C+) may serve as a therapeutic approach for treating inflammatory disorders.",
keywords = "IBD, Immunomodulation, IRF8, LNP, Nanoparticle, RNAi",
author = "Nuphar Veiga and Meir Goldsmith and Yael Diesendruck and Srinivas Ramishetti and Daniel Rosenblum and Eran Elinav and Behlke, {Mark A.} and Itai Benhar and Dan Peer",
year = "2019",
month = "11",
day = "10",
doi = "10.1016/j.jconrel.2019.10.001",
language = "English (US)",
volume = "313",
pages = "33--41",
journal = "Journal of Controlled Release",
issn = "0168-3659",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Leukocyte-specific siRNA delivery revealing IRF8 as a potential anti-inflammatory target

AU - Veiga, Nuphar

AU - Goldsmith, Meir

AU - Diesendruck, Yael

AU - Ramishetti, Srinivas

AU - Rosenblum, Daniel

AU - Elinav, Eran

AU - Behlke, Mark A.

AU - Benhar, Itai

AU - Peer, Dan

PY - 2019/11/10

Y1 - 2019/11/10

N2 - Interferon regulatory factor 8 (IRF8) protein plays a critical role in the differentiation, polarization, and activation of mononuclear phagocytic cells. In light of previous studies, we explored the therapeutic potential of IRF8 inhibition as immunomodulatory therapy for inflammatory bowel disease (IBD). To this end, we utilized siRNA-loaded lipid-based nanoparticles (siLNPs) and demonstrated a ∼90% reduction of IRF8 mRNA levels in vitro (PV < 0.0001), alongside a notable reduction in IRF8 protein. Moreover, silencing IRF8 ex vivo in splenocytes lead to a profound downregulation of IRF8 protein, followed by an immunomodulatory effect, as represented by a decrease in the secretion of TNFα, IL6 and IL12/IL23 (IL12p40) proinflammatory cytokines (PV = 0.0045, 0.0330, <0.0001, respectively). In order to silence IRF8 in vivo, selectively in inflammatory leukocytes, we used siLNPs that were coated with anti-Ly6C antibodies via our recently published ASSET targeting approach. Through this strategy, we have demonstrated a selective binding of the targeted-LNPs (T-LNPs) to Ly6C + inflammatory leukocytes. Finally, an immunomodulatory effect was demonstrated in vivo in an IBD mouse model with a profound decrease of TNFα, IL6, IL12/IL23, and IL1β pro-inflammatory cytokines (n = 5, PV < 0.0001, <0.0001, <0.0001, 0.02, respectively) and an improvement of colon-morphology as assessed by colon-length measurements and colonoscopy (PV < 0.0001). Overall, using antibody-targeted siLNPs, we showed a notable reduction of IRF8 mRNA and protein and demonstrated a targeted immunomodulation therapeutic effect ex vivo and in vivo, in the DSS colitis model. We claim that a selective silencing of IRF8 in inflammatory leukocytes (such as Ly6C+) may serve as a therapeutic approach for treating inflammatory disorders.

AB - Interferon regulatory factor 8 (IRF8) protein plays a critical role in the differentiation, polarization, and activation of mononuclear phagocytic cells. In light of previous studies, we explored the therapeutic potential of IRF8 inhibition as immunomodulatory therapy for inflammatory bowel disease (IBD). To this end, we utilized siRNA-loaded lipid-based nanoparticles (siLNPs) and demonstrated a ∼90% reduction of IRF8 mRNA levels in vitro (PV < 0.0001), alongside a notable reduction in IRF8 protein. Moreover, silencing IRF8 ex vivo in splenocytes lead to a profound downregulation of IRF8 protein, followed by an immunomodulatory effect, as represented by a decrease in the secretion of TNFα, IL6 and IL12/IL23 (IL12p40) proinflammatory cytokines (PV = 0.0045, 0.0330, <0.0001, respectively). In order to silence IRF8 in vivo, selectively in inflammatory leukocytes, we used siLNPs that were coated with anti-Ly6C antibodies via our recently published ASSET targeting approach. Through this strategy, we have demonstrated a selective binding of the targeted-LNPs (T-LNPs) to Ly6C + inflammatory leukocytes. Finally, an immunomodulatory effect was demonstrated in vivo in an IBD mouse model with a profound decrease of TNFα, IL6, IL12/IL23, and IL1β pro-inflammatory cytokines (n = 5, PV < 0.0001, <0.0001, <0.0001, 0.02, respectively) and an improvement of colon-morphology as assessed by colon-length measurements and colonoscopy (PV < 0.0001). Overall, using antibody-targeted siLNPs, we showed a notable reduction of IRF8 mRNA and protein and demonstrated a targeted immunomodulation therapeutic effect ex vivo and in vivo, in the DSS colitis model. We claim that a selective silencing of IRF8 in inflammatory leukocytes (such as Ly6C+) may serve as a therapeutic approach for treating inflammatory disorders.

KW - IBD

KW - Immunomodulation

KW - IRF8

KW - LNP

KW - Nanoparticle

KW - RNAi

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UR - http://www.scopus.com/inward/citedby.url?scp=85073962267&partnerID=8YFLogxK

U2 - 10.1016/j.jconrel.2019.10.001

DO - 10.1016/j.jconrel.2019.10.001

M3 - Article

VL - 313

SP - 33

EP - 41

JO - Journal of Controlled Release

T2 - Journal of Controlled Release

JF - Journal of Controlled Release

SN - 0168-3659

ER -

ID: 54647562