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Off-target hepatic protein expression from delivered mRNA raises safety concerns, and hepatic clearance limits efficient iLNP delivery to target organs. In this study, we address these challenges in mouse models by stealth-coating the liver sinusoidal endothelial (LSE) wall, the primary gateway for nanoparticle entry into the liver. Specifically, oligocations conjugated with two-armed PEG (2-arm-PEG-oligocations), a clinically relevant material used in oligonucleotide delivery trials, were employed to transiently anchor PEG to the LSE wall with balanced affinity, ensuring robust coating followed by gradual biliary clearance. This approach reduced hepatic protein expression from iLNPs, subsequently administered either systemically or locally, by more than 10-fold. Importantly, the strategy preserved iLNP accumulation in the spleen, a key target organ for vaccines, effectively redirecting iLNPs from the liver to the spleen. Consequently, in vaccine applications, preinjection of the 2-arm-PEG-oligocation preserved or even enhanced vaccination efficacy while minimizing concerns associated with antigen expression in the liver. In applications involving cytokine mRNA therapy, specifically intratumoral interleukin-12 (IL-12) mRNA administration, systemic preinjection of the 2-arm-PEG-oligocation successfully reduced off-target hepatic IL-12 expression and subsequent systemic IL-12 exposure, while maintaining antitumor efficacy. Collectively, these results demonstrate that LSE-wall stealth coating is a generalizable strategy to improve both the safety and efficacy of iLNP-based mRNA vaccines and immunotherapies.<\/jats:p>","DOI":"10.1021\/acsnano.6c08403","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T06:02:19Z","timestamp":1787032939000},"page":"23958-23974","source":"Crossref","is-referenced-by-count":1,"title":["2-arm-PEG-Oligocations\nTransiently Shield the\nLiver Sinusoids to Mitigate Off-Target Hepatic Expression of mRNA\nLipid Nanoparticles"],"prefix":"10.1021","volume":"20","author":[{"given":"Anjaneyulu","family":"Dirisala","sequence":"first","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0288-0921","authenticated-orcid":true,"given":"Bhaskar","family":"Chatterjee","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"Institute of Science Tokyo , , 1-5-45 Yushima , Bunkyo-Ku, ,","place":["Tokyo, Japan, 113-8510"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nguyen Bui Thao","family":"Le","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"Institute of Science Tokyo , , 1-5-45 Yushima , Bunkyo-Ku, ,","place":["Tokyo, Japan, 113-8510"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuko","family":"Toh","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miki","family":"Matsui-Masai","sequence":"additional","affiliation":[{"name":"NANO MRNA Co. Ltd. 2-5-1 Atago , Minato-Ku, ,","place":["Tokyo, Japan, 105-6226"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8874-233X","authenticated-orcid":true,"given":"Saed","family":"Abbasi","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"The Johns Hopkins University School of Medicine , , 400 N. Broadway , , ,","place":["Baltimore, Maryland, United States, 21231"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xueying","family":"Liu","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2736-2058","authenticated-orcid":true,"given":"Theofilus A.","family":"Tockary","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nan","family":"Qiao","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"Institute of Science Tokyo , , 1-5-45 Yushima , Bunkyo-Ku, ,","place":["Tokyo, Japan, 113-8510"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junichi","family":"Ishikawa","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"Institute of Science Tokyo , , 1-5-45 Yushima , Bunkyo-Ku, ,","place":["Tokyo, Japan, 113-8510"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4857-8825","authenticated-orcid":true,"given":"Jumpei","family":"Norimatsu","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"Institute of Science Tokyo , , 1-5-45 Yushima , Bunkyo-Ku, ,","place":["Tokyo, Japan, 113-8510"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5559-8519","authenticated-orcid":true,"given":"Yuki","family":"Mochida","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"Institute of Science Tokyo , , 1-5-45 Yushima , Bunkyo-Ku, ,","place":["Tokyo, Japan, 113-8510"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shigeto","family":"Fukushima","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3691-3608","authenticated-orcid":true,"given":"Makoto","family":"Oba","sequence":"additional","affiliation":[{"name":"Kyoto Prefectural University of Medicine , , 1-5 Shimogamohangi-Cho , Sakyo-Ku, ,","place":["Kyoto, Japan, 606-0823"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazunori","family":"Kataoka","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8726-0478","authenticated-orcid":true,"given":"Satoshi","family":"Uchida","sequence":"additional","affiliation":[{"name":"Kawasaki Institute of Industrial Promotion , , 3-25-14 Tonomachi , Kawasaki-Ku, ,","place":["Kawasaki, Japan, 210-0821"]},{"name":"Institute of Science Tokyo , , 1-5-45 Yushima , Bunkyo-Ku, ,","place":["Tokyo, Japan, 113-8510"]},{"name":"Kyoto Prefectural University of Medicine , , 1-5 Shimogamohangi-Cho , Sakyo-Ku, ,","place":["Kyoto, Japan, 606-0823"]},{"name":"The University of Tokyo , , 4-6-1, Shirokanedai , Minato-Ku, ,","place":["Tokyo, Japan, 108-0071"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,18]]},"reference":[{"issue":"12","key":"2026090306595593700_cit1","doi-asserted-by":"publisher","first-page":"1078","DOI":"10.1056\/NEJMoa2110475","article-title":"Safety of the BNT162b2 mRNA Covid-19 vaccine in a nationwide\nsetting","volume":"385","author":"Barda","year":"2021","journal-title":"N. 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Ther."},{"issue":"2","key":"2026090306595593700_cit5","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1016\/j.immuni.2005.06.008","article-title":"Suppression of RNA\nrecognition by Toll-like receptors: the impact of nucleoside modification\nand the evolutionary origin of RNA","volume":"23","author":"Kariko","year":"2005","journal-title":"Immunity"},{"key":"2026090306595593700_cit6","doi-asserted-by":"publisher","first-page":"337","DOI":"10.1016\/j.jconrel.2015.08.051","article-title":"N(1)-methylpseudouridine-incorporated mRNA outperforms\npseudouridine-incorporated mRNA by providing enhanced protein expression\nand reduced immunogenicity in mammalian cell lines and mice","volume":"217","author":"Andries","year":"2015","journal-title":"J. Controlled Release"},{"issue":"5","key":"2026090306595593700_cit7","doi-asserted-by":"publisher","first-page":"748","DOI":"10.1021\/acscentsci.1c00197","article-title":"Modifications in an emergency: The\nrole of N1-methylpseudouridine\nin COVID-19 vaccines","volume":"7","author":"Nance","year":"2021","journal-title":"ACS Cent. Sci."},{"issue":"12","key":"2026090306595593700_cit8","doi-asserted-by":"publisher","first-page":"1078","DOI":"10.1038\/s41578-021-00358-0","article-title":"Lipid nanoparticles\nfor mRNA delivery","volume":"6","author":"Hou","year":"2021","journal-title":"Nat. Rev. Mater."},{"issue":"16","key":"2026090306595593700_cit9","doi-asserted-by":"publisher","first-page":"15231","DOI":"10.1021\/acsnano.2c12584","article-title":"The mRNA vaccine revolution:\nCOVID-19 has launched the future of\nvaccinology","volume":"17","author":"Ye","year":"2023","journal-title":"ACS Nano"},{"key":"2026090306595593700_cit10","doi-asserted-by":"publisher","first-page":"114972","DOI":"10.1016\/j.addr.2023.114972","article-title":"Polyplex designs for\nimproving the stability and safety of RNA therapeutics","volume":"199","author":"Uchida","year":"2023","journal-title":"Adv. Drug Delivery Rev."},{"issue":"11","key":"2026090306595593700_cit11","doi-asserted-by":"publisher","first-page":"1993","DOI":"10.1016\/j.immuni.2022.10.014","article-title":"Innate immune mechanisms\nof mRNA vaccines","volume":"55","author":"Verbeke","year":"2022","journal-title":"Immunity"},{"issue":"1","key":"2026090306595593700_cit12","doi-asserted-by":"publisher","first-page":"422","DOI":"10.1080\/15476286.2024.2333123","article-title":"S. mRNA vaccine designs\nfor optimal adjuvanticity and\ndelivery","volume":"21","author":"Mochida","year":"2024","journal-title":"RNA Biol."},{"issue":"12","key":"2026090306595593700_cit13","doi-asserted-by":"publisher","first-page":"2877","DOI":"10.1016\/j.immuni.2021.11.001","article-title":"Lipid nanoparticles enhance the efficacy of mRNA and\nprotein subunit vaccines by inducing robust T follicular helper cell\nand humoral responses","volume":"54","author":"Alameh","year":"2021","journal-title":"Immunity"},{"key":"2026090306595593700_cit14","doi-asserted-by":"publisher","first-page":"474","DOI":"10.1038\/s41590-022-01168-4","article-title":"Making innate sense\nof mRNA vaccine adjuvanticity","volume":"23","author":"Kobiyama","year":"2022","journal-title":"Nat. Immunol."},{"issue":"5","key":"2026090306595593700_cit15","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1038\/s41551-019-0378-3","article-title":"Visualization\nof early\nevents in mRNA vaccine delivery in non-human primates via PET-CT and\nnear-infrared imaging","volume":"3","author":"Lindsay","year":"2019","journal-title":"Nat. Biomed. Eng."},{"issue":"1","key":"2026090306595593700_cit16","doi-asserted-by":"publisher","first-page":"102083","DOI":"10.1016\/j.omtn.2023.102083","article-title":"E. mRNA vaccine trafficking and resulting\nprotein expression after intramuscular administration","volume":"35","author":"Hassett","year":"2024","journal-title":"Mol. Ther."},{"issue":"9","key":"2026090306595593700_cit17","doi-asserted-by":"publisher","first-page":"3078","DOI":"10.1016\/j.ymthe.2022.07.007","article-title":"mRNA-LNP vaccines tuned\nfor systemic immunization induce strong antitumor immunity by engaging\nsplenic immune cells","volume":"30","author":"Bevers","year":"2022","journal-title":"Mol. Ther."},{"issue":"45","key":"2026090306595593700_cit18","doi-asserted-by":"publisher","first-page":"2303795","DOI":"10.1002\/adfm.202303795","article-title":"Harnessing topology and stereochemistry\nof glycidylamine-derived lipid nanoparticles for in vivo mRNA delivery\nto immune cells in spleen and their application for cancer vaccination","volume":"33","author":"Abd Elwakil","year":"2023","journal-title":"Adv. Funct. Mater."},{"key":"2026090306595593700_cit19","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1016\/j.jconrel.2023.03.041","article-title":"Spleen-selective\nco-delivery of mRNA and TLR4 agonist-loaded LNPs\nfor synergistic immunostimulation and Th1 immune responses","volume":"357","author":"Pan","year":"2023","journal-title":"J. Controlled Release"},{"issue":"46","key":"2026090306595593700_cit20","doi-asserted-by":"publisher","DOI":"10.1002\/adma.202303614","article-title":"Esterase-labile quaternium lipidoid enabling improved\nmRNA-LNP stability and spleen-selective mRNA transfection","volume":"35","author":"Zhang","year":"2023","journal-title":"Adv. Mater."},{"issue":"44","key":"2026090306595593700_cit21","doi-asserted-by":"publisher","first-page":"30701","DOI":"10.1021\/acsnano.4c09902","article-title":"Spleen-targeted mRNA vaccine doped\nwith manganese adjuvant for robust anticancer immunity in vivo","volume":"18","author":"Luo","year":"2024","journal-title":"ACS Nano"},{"issue":"41","key":"2026090306595593700_cit22","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.ady6460","article-title":"Replacing cholesterol and PEGylated lipids\nwith zwitterionic ionizable\nlipids in LNPs for spleen-specific mRNA translation","volume":"11","author":"Zhao","year":"2025","journal-title":"Sci. Adv."},{"key":"2026090306595593700_cit23","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1016\/j.addr.2020.06.019","article-title":"Controlling timing and location in\nvaccines","volume":"158","author":"Irvine","year":"2020","journal-title":"Adv. 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Ther."},{"key":"2026090306595593700_cit27","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1016\/j.jconrel.2020.03.006","article-title":"Different\nkinetics for the hepatic uptake of lipid nanoparticles between the\napolipoprotein E\/low density lipoprotein receptor and the N-acetyl-d-galactosamine\/asialoglycoprotein\nreceptor pathway","volume":"322","author":"Sato","year":"2020","journal-title":"J. Controlled Release"},{"issue":"52","key":"2026090306595593700_cit28","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2109256118","article-title":"On the\nmechanism of tissue-specific\nmRNA delivery by selective organ targeting nanoparticles","volume":"118","author":"Dilliard","year":"2021","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"4","key":"2026090306595593700_cit29","doi-asserted-by":"publisher","first-page":"1751","DOI":"10.1002\/j.2040-4603.2015.tb00660.x","article-title":"Liver sinusoidal\nendothelial\ncells","volume":"5","author":"Sorensen","year":"2015","journal-title":"Compr. Physiol."},{"issue":"3","key":"2026090306595593700_cit30","doi-asserted-by":"publisher","first-page":"2138","DOI":"10.1021\/acsnano.7b06995","article-title":"Directing\nnanoparticle biodistribution through evasion and exploitation of stab2-dependent\nnanoparticle uptake","volume":"12","author":"Campbell","year":"2018","journal-title":"ACS Nano"},{"issue":"10","key":"2026090306595593700_cit31","doi-asserted-by":"publisher","first-page":"10130","DOI":"10.1021\/acsnano.8b04947","article-title":"Dense and dynamic polyethylene\nglycol shells cloak nanoparticles from uptake by liver endothelial\ncells for long blood circulation","volume":"12","author":"Zhou","year":"2018","journal-title":"ACS Nano"},{"issue":"11","key":"2026090306595593700_cit32","doi-asserted-by":"publisher","first-page":"1212","DOI":"10.1038\/nmat4718","article-title":"Mechanism of hard-nanomaterial clearance by\nthe liver","volume":"15","author":"Tsoi","year":"2016","journal-title":"Nat. Mater."},{"key":"2026090306595593700_cit33","doi-asserted-by":"publisher","first-page":"345","DOI":"10.1016\/j.jconrel.2015.08.007","article-title":"Expression\nkinetics of nucleoside-modified mRNA delivered in lipid nanoparticles\nto mice by various routes","volume":"217","author":"Pardi","year":"2015","journal-title":"J. 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