전립선암 표적을 위한 hCD64 단백질 발현 세포막 유래 나노 소포 연구

Abstract

Extracellular vesicles (EVs) are ideal therapeutic and diagnostic tools for various diseases. However, EVs commonly lack mass production and targeting capability, and are easily phagocytosed by immune cells such as macrophages. This requires their administration at high doses, which can lead to toxicity and adverse reactions. To overcome these limitations, the use of cell-derived mimetic nanovesicles containing Fc gamma receptor (FcγR) protein was proposed in this study. Plasma membrane-derived nanovesicles are fabricated with blebs produced in giant plasma membrane vesicles (GPMVs) through the treatment of sulfide breaker, N-Ethylmaleimide (NEM). The plasma membrane-derived nanovesicles are named bleb-derived nanovesicles (BNVs). BNVs are derived from cells that have low toxicity and immunogenicity. Their homogeneous nano-scale dimensions allow for enhancing tissue penetration. Furthermore, the internal components of BNVs lack various organelles, thereby minimizing the potential risk. In this study, BNVs were engineered from HEK293T cells expressing human FcγRI (hCD64), which has the strongest affinity to monomeric IgG among the FcR family. To enhance prostate cancer targeting, anti-PSMA (prostate-specific membrane antigen) antibodies were bound to the hCD64-expressing-BNVs and confirmed their targeting ability. This study verified the expression and functionality of hCD64 protein at the cellular and BNV levels. We determined concentration and time conditions for optimal binding and validated targeting ability using PSMA-expressing cell lines. To confirm in vivo targeting, the prostate cancer xenograft mouse model was established using 22Rv1, a human prostate cancer cell line. It was proved that the target function was possible even in vivo through IgG2 attached hCD64-BNVs (hCD64- BNVs-IgG2). This study demonstrated the potential of hCD64-expressing-BNVs as a platform for the diagnosis and therapy of prostate cancer by confirming their ability to target PSMA. Therefore, this study is expected to contribute to the proposition of “precision targeting” for cancer diagnosis and therapy systems. The hypothesis and workflow for this study are illustrated in scheme 1. Scheme 1. Bleb-derived nanovesicles (BNVs) expressing human FcγRI (hCD64) protein enhanced the targeting ability of prostate cancer.