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基因表面修饰的外膜囊泡靶向癌细胞中的MUC1抗原。

Genetically surface-modified outer membrane vesicles targeting MUC1 antigen in cancer cells.

作者信息

Laotee Sedthawut, Arunmanee Wanatchaporn

机构信息

Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.

Center of Excellence in Cancer Cell and Molecular Biology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.

出版信息

Biotechnol Rep (Amst). 2024 Sep 3;44:e00854. doi: 10.1016/j.btre.2024.e00854. eCollection 2024 Dec.

DOI:10.1016/j.btre.2024.e00854
PMID:39290790
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11406022/
Abstract

Outer membrane vesicles (OMVs), non-replicating spherical liposomes derived from Gram-negative bacteria, are a promising vaccine platform and multifunctional delivery systems. Their ability to be modified via genetic engineering for the incorporation and display of heterologous proteins enhances their functionality. In this study, we demonstrated a bio-ligation approach to display single-chain variable fragments (scFv) on the OMV surface using the SpyTag/SpyCatcher system. SpyTag-fused scFv, expressed by mammalian cells, bound to OMVs with SpyCatcher-fused Lpp'OmpA after a simple incubation. Biophysical analysis indicated that the conjugated OMVs maintained their physicochemical properties. We used an scFv targeting mucin 1 protein (MUC1) for specific cell targeting. Confocal microscopy revealed that conjugated OMVs specifically bound to and were internalized by MUC1-presenting cells, but not by MUC1-deficient cells. In conclusion, this rapid and efficient bio-ligation system facilitates the display of functional scFv on OMV surfaces, offering a promising approach for targeted delivery to MUC1-expressing cancer cells.

摘要

外膜囊泡（OMV）是源自革兰氏阴性菌的非复制性球形脂质体，是一种很有前景的疫苗平台和多功能递送系统。它们能够通过基因工程进行修饰，以掺入和展示异源蛋白，从而增强其功能。在本研究中，我们展示了一种生物连接方法，即使用SpyTag/SpyCatcher系统在外膜囊泡表面展示单链可变片段（scFv）。由哺乳动物细胞表达的与SpyTag融合的scFv，在简单孵育后与与SpyCatcher融合的Lpp'OmpA结合到外膜囊泡上。生物物理分析表明，缀合的外膜囊泡保持了它们的物理化学性质。我们使用靶向粘蛋白1蛋白（MUC1）的scFv进行特异性细胞靶向。共聚焦显微镜显示，缀合的外膜囊泡特异性结合到表达MUC1的细胞并被其内化，但不会被缺乏MUC1的细胞内化。总之，这种快速有效的生物连接系统有助于在外膜囊泡表面展示功能性scFv，为靶向递送至表达MUC1的癌细胞提供了一种很有前景的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/814861ad5750/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/4f3ee682f7cb/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/e01de373baf5/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/31823eca65db/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/983d834134f9/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/92743c25d851/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/2174583969e1/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/814861ad5750/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/4f3ee682f7cb/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/e01de373baf5/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/31823eca65db/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/983d834134f9/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/92743c25d851/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/2174583969e1/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ee3/11406022/814861ad5750/gr7.jpg

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