{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T18:46:14Z","timestamp":1792608374267,"version":"4.5.0"},"reference-count":48,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2022,4,16]],"date-time":"2022-04-16T00:00:00Z","timestamp":1650067200000},"content-version":"vor","delay-in-days":15,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001352","name":"National University of Singapore","doi-asserted-by":"publisher","award":["NUHSRO\/2019\/076\/STARTUP\/02"],"award-info":[{"award-number":["NUHSRO\/2019\/076\/STARTUP\/02"]}],"id":[{"id":"10.13039\/501100001352","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Extracellular Vesicle"],"published-print":{"date-parts":[[2022,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The RIG\u2010I pathway can be activated by RNA containing 5\u2032 triphosphate, leading to type I interferon release and immune activation. Hence, RIG\u2010I agonists have been used to induce immune responses against cancer as potential immunotherapy. However, delivery of 5\u2032 triphosphorylated RNA molecules as RIG\u2010I agonists to tumour cells in vivo is challenging due to the susceptibility of these molecules to degradation. In this study, we demonstrate the use of extracellular vesicles (EVs) from red blood cells (RBCs), which are highly amenable for RNA loading and taken up robustly by cancer cells, for RIG\u2010I agonist delivery. We evaluate the anti\u2010cancer activity of two novel RIG\u2010I agonists, the immunomodulatory RNA (immRNA) with a unique secondary structure for efficient RIG\u2010I activation, and a 5\u2032 triphosphorylated antisense oligonucleotide with dual function of RIG\u2010I activation and miR\u2010125b inhibition (3p\u2010125b\u2010ASO). We find that RBCEV\u2010delivered immRNA and 3p\u2010125b\u2010ASO trigger the RIG\u2010I pathway, and induce cell death in both mouse and human breast cancer cells. Furthermore, we observe a significant suppression of tumour growth coupled with increased immune cell infiltration mediated by the activation of RIG\u2010I cascade after multiple intratumoral injections of RBCEVs loaded with immRNA or 3p\u2010125b\u2010ASO. Targeted delivery of immRNA using RBCEVs with EGFR\u2010binding nanobody administrated via intrapulmonary delivery facilitates the accumulation of RBCEVs in metastatic cancer cells, leading to potent tumour\u2010specific CD8<jats:sup>+<\/jats:sup> T cells immune response. This contributes to prominent suppression of breast cancer metastasis in the lung. Hence, this study provides a new strategy for efficient RIG\u2010I agonist delivery using RBCEVs for immunotherapy against cancer and cancer metastasis.<\/jats:p>","DOI":"10.1002\/jev2.12187","type":"journal-article","created":{"date-parts":[[2022,4,17]],"date-time":"2022-04-17T00:30:25Z","timestamp":1650155425000},"source":"Crossref","is-referenced-by-count":84,"title":["Robust delivery of RIG\u2010I agonists using extracellular vesicles for anti\u2010cancer immunotherapy"],"prefix":"10.1002","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9956-0889","authenticated-orcid":false,"given":"Boya","family":"Peng","sequence":"first","affiliation":[{"name":"Department of Pharmacology and Institute for Digital Medicine Yong Loo Lin School of Medicine National University of Singapore  Singapore"},{"name":"Department of Surgery Immunology Program Cancer Program and Nanomedicine Translational Program Yong Loo Lin School of Medicine National University of Singapore  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