Page 86 - 2025中醫藥與天然藥物聯合學術研討會-中醫藥與天然藥物的挑戰Ｘ機遇與未來大會手冊

Dual-Targeted Extracellular Vesicles as Natural Nanocarriers for Multi-

Gene Therapy in Pancreatic Cancer

1
1
Bi-Da Hsiang, Guan-Wan Liu, Chi-Ling Chiang* ,1,2

1 Institute of Biopharmaceutical Sciences, National Yang Ming Chiao Tung University, Taipei,
11221 Taiwan
2 Department of Chemical and Biomolecular Engineering, The Ohio State University,
Columbus, OH, 43210 USA
* E-mail: Clchiang@nycu.edu.tw

Abstract
Extracellular vesicles (EVs), including exosomes and microvesicles, have emerged as
attractive alternatives for gene delivery because of their low immunogenicity, minimal
cytotoxicity, and ability to cross physiological barriers. As naturally derived carriers secreted
by cells, EVs can themselves be regarded as a form of natural medicine, uniquely suited for
therapeutic delivery. Yet, EV-based therapy still faces major challenges in large-scale
production, selective targeting, and efficient cargo encapsulation. Pancreatic ductal
adenocarcinoma (PDAC), a malignancy marked by multiple gene mutations, dense stromal
fibrosis, and poor therapeutic response, highlights the urgent need for carriers capable of
delivering multiple gene cargoes to bulky solid tumors. To address this, our group developed a
dual-targeted extracellular vesicle (dtEV) engineered to carry high loads of therapeutic mRNA
and siRNA. These dtEVs display an Fc receptor anchor protein coupled with a tissue-targeting
peptide and a humanized monoclonal antibody, enabling precise tumor targeting. Using an
asymmetric cell electroporation platform, we sequentially transfected donor cells with plasmids
encoding both the RNA and protein of interest, resulting in abundant dtEVs with high RNA
loading efficiency.
In preclinical models, including fibrotic PDAC tumoroids that recapitulate the desmoplastic
tumor microenvironment, as well as orthotopic PANC-1 and patient-derived xenograft tumors,
dtEVs carrying KRAS G12D siRNA and TP53 mRNA, when combined with low-dose
Gemcitabine, significantly suppressed tumor growth and metastasis while prolonging survival.
Our work demonstrates a clinically accessible and scalable strategy for producing targeted EVs,
positioning them as a next-generation natural medicine for delivering multiple therapeutic gene
cargoes to large, treatment-resistant, and fibrotic solid tumors.

Keywords: Extracellular vesicles (EVs); Dual-targeting; RNA delivery; Pancreatic ductal
adenocarcinoma (PDAC); Precision oncology

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