Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy
The RIG-I pathway can be activated by RNA containing 5' triphosphate, leading to type I interferon release and immune activation. Hence, RIG-I agonists have been used to induce immune responses against cancer as potential immunotherapy. However, delivery of 5' triphosphorylated RNA molecul...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2022
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/163208 |
| _version_ | 1824456123884765184 |
|---|---|
| author | Peng, Boya Nguyen, Trinh Mai Jayasinghe, Migara Kavishka Gao, Chang Pham, Thach Tuan Vu, Luyen Tien Yeo, Eric Yew Meng Yap, Gracemary Wang, Lingzhi Goh, Boon Cher Tam, Wai Leong Luo, Dahai Le, Minh TN |
| author2 | Lee Kong Chian School of Medicine (LKCMedicine) |
| author_facet | Lee Kong Chian School of Medicine (LKCMedicine) Peng, Boya Nguyen, Trinh Mai Jayasinghe, Migara Kavishka Gao, Chang Pham, Thach Tuan Vu, Luyen Tien Yeo, Eric Yew Meng Yap, Gracemary Wang, Lingzhi Goh, Boon Cher Tam, Wai Leong Luo, Dahai Le, Minh TN |
| author_sort | Peng, Boya |
| collection | NTU |
| description | The RIG-I pathway can be activated by RNA containing 5' triphosphate, leading to type I interferon release and immune activation. Hence, RIG-I agonists have been used to induce immune responses against cancer as potential immunotherapy. However, delivery of 5' triphosphorylated RNA molecules as RIG-I 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-I agonist delivery. We evaluate the anti-cancer activity of two novel RIG-I agonists, the immunomodulatory RNA (immRNA) with a unique secondary structure for efficient RIG-I activation, and a 5' triphosphorylated antisense oligonucleotide with dual function of RIG-I activation and miR-125b inhibition (3p-125b-ASO). We find that RBCEV-delivered immRNA and 3p-125b-ASO trigger the RIG-I 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-I cascade after multiple intratumoral injections of RBCEVs loaded with immRNA or 3p-125b-ASO. Targeted delivery of immRNA using RBCEVs with EGFR-binding nanobody administrated via intrapulmonary delivery facilitates the accumulation of RBCEVs in metastatic cancer cells, leading to potent tumour-specific CD8+ 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-I agonist delivery using RBCEVs for immunotherapy against cancer and cancer metastasis. |
| first_indexed | 2025-02-19T03:49:06Z |
| format | Journal Article |
| id | ntu-10356/163208 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2025-02-19T03:49:06Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | ntu-10356/1632082023-03-05T16:52:39Z Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy Peng, Boya Nguyen, Trinh Mai Jayasinghe, Migara Kavishka Gao, Chang Pham, Thach Tuan Vu, Luyen Tien Yeo, Eric Yew Meng Yap, Gracemary Wang, Lingzhi Goh, Boon Cher Tam, Wai Leong Luo, Dahai Le, Minh TN Lee Kong Chian School of Medicine (LKCMedicine) NTU Institute of Structural Biology Science::Medicine Cancer Extracellular Vesicles, The RIG-I pathway can be activated by RNA containing 5' triphosphate, leading to type I interferon release and immune activation. Hence, RIG-I agonists have been used to induce immune responses against cancer as potential immunotherapy. However, delivery of 5' triphosphorylated RNA molecules as RIG-I 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-I agonist delivery. We evaluate the anti-cancer activity of two novel RIG-I agonists, the immunomodulatory RNA (immRNA) with a unique secondary structure for efficient RIG-I activation, and a 5' triphosphorylated antisense oligonucleotide with dual function of RIG-I activation and miR-125b inhibition (3p-125b-ASO). We find that RBCEV-delivered immRNA and 3p-125b-ASO trigger the RIG-I 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-I cascade after multiple intratumoral injections of RBCEVs loaded with immRNA or 3p-125b-ASO. Targeted delivery of immRNA using RBCEVs with EGFR-binding nanobody administrated via intrapulmonary delivery facilitates the accumulation of RBCEVs in metastatic cancer cells, leading to potent tumour-specific CD8+ 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-I agonist delivery using RBCEVs for immunotherapy against cancer and cancer metastasis. Ministry of Education (MOE) Ministry of Health (MOH) National Medical Research Council (NMRC) Published version This project is funded by the National University of Singapore (grant number NUHSRO/2019/076/STARTUP/02), and the Singapore Ministry of Education (NUHSRO/2020/108/T1/Seed-Mar/04). Gracemary Yap is supported by the NUS Resilient and Growth fellowship (NUS-13), financed by the National Science Foundation of Singapore. This research is also supported by the Singapore Ministry of Health’s National Medical Research Council under its Open Fund – Individual Research Grant (NMRC/OFIRG/0075/2018and OFIRG20nov-0049). 2022-11-29T01:28:14Z 2022-11-29T01:28:14Z 2022 Journal Article Peng, B., Nguyen, T. M., Jayasinghe, M. K., Gao, C., Pham, T. T., Vu, L. T., Yeo, E. Y. M., Yap, G., Wang, L., Goh, B. C., Tam, W. L., Luo, D. & Le, M. T. (2022). Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy. Journal of Extracellular Vesicles, 11(4), e12187-. https://dx.doi.org/10.1002/jev2.12187 2001-3078 https://hdl.handle.net/10356/163208 10.1002/jev2.12187 35430766 2-s2.0-85128386325 4 11 e12187 en NUHSRO/2019/076/STARTUP/02 NUHSRO/2020/108/T1/Seed-Mar/04 NMRC/OFIRG/0075/2018and OFIRG20nov-0049 Journal of Extracellular Vesicles © 2022 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. application/pdf |
| spellingShingle | Science::Medicine Cancer Extracellular Vesicles, Peng, Boya Nguyen, Trinh Mai Jayasinghe, Migara Kavishka Gao, Chang Pham, Thach Tuan Vu, Luyen Tien Yeo, Eric Yew Meng Yap, Gracemary Wang, Lingzhi Goh, Boon Cher Tam, Wai Leong Luo, Dahai Le, Minh TN Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy |
| title | Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy |
| title_full | Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy |
| title_fullStr | Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy |
| title_full_unstemmed | Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy |
| title_short | Robust delivery of RIG-I agonists using extracellular vesicles for anti-cancer immunotherapy |
| title_sort | robust delivery of rig i agonists using extracellular vesicles for anti cancer immunotherapy |
| topic | Science::Medicine Cancer Extracellular Vesicles, |
| url | https://hdl.handle.net/10356/163208 |
| work_keys_str_mv | AT pengboya robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT nguyentrinhmai robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT jayasinghemigarakavishka robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT gaochang robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT phamthachtuan robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT vuluyentien robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT yeoericyewmeng robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT yapgracemary robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT wanglingzhi robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT gohbooncher robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT tamwaileong robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT luodahai robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy AT leminhtn robustdeliveryofrigiagonistsusingextracellularvesiclesforanticancerimmunotherapy |