Chen Mei-Yi, Cheng Ting-Wei, Pan Yi-Chung, Mou Chung-Yuan, Chiang Yun-Wei, Lin Wan-Chen, Hu Che-Ming Jack, Mou Kurt Yun
Chemical Biology and Molecular Biophysics Program, Taiwan International Graduate Program, Academia Sinica, No. 128, Sec. 2, Academia Rd., Nangang (Nankang) Dist., Taipei City 115201, Taiwan.
Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan.
ACS Synth Biol. 2025 Jan 17;14(1):148-160. doi: 10.1021/acssynbio.4c00483. Epub 2025 Jan 6.
Bacterial outer membrane vesicles (OMVs) have emerged as promising vehicles for anticancer drug delivery due to their inherent tumor tropism, immune-stimulatory properties, and potential for functionalization with therapeutic proteins. Despite their advantages, the high lipopolysaccharide (LPS) endotoxin content in the OMVs raises significant safety and regulatory challenges. In this work, we produce LPS-attenuated and LPS-free OMVs and systematically assess the effects of LPS modification on OMVs' physicochemical characteristics, membrane protein content, immune-stimulatory capacity, tolerability, and anticancer efficacy. Our findings reveal that LPS removal increased the maximal tolerated dose of the OMVs by over 25-fold. When adjusted for comparable safety profiles, LPS-free OMVs exhibit superior anticancer effects compared with wild-type OMVs. Mechanistic investigations indicate that the LPS removal obviates immune cell death caused by LPS and reduces the negatory effects of wild type of OMVs on tumor immune cell infiltrates. We further show the functionality of the LPS-free OMV through the incorporation of an IL-2 variant protein (Neo-2/15). This functionalization augments OMV's ability of the OMV to inhibit tumor growth and promote lymphocyte infiltration into the tumor microenvironment. This study presents a safe and functionalizable OMV with improved translational prospect.
细菌外膜囊泡（OMV）因其固有的肿瘤趋向性、免疫刺激特性以及与治疗性蛋白质功能化的潜力，已成为抗癌药物递送的有前景的载体。尽管具有这些优势，但OMV中高含量的脂多糖（LPS）内毒素带来了重大的安全和监管挑战。在这项工作中，我们制备了LPS减弱型和无LPS的OMV，并系统评估了LPS修饰对OMV的物理化学特性、膜蛋白含量、免疫刺激能力、耐受性和抗癌功效的影响。我们的研究结果表明，去除LPS可使OMV的最大耐受剂量增加超过25倍。在调整为可比的安全概况后，无LPS的OMV与野生型OMV相比表现出更优异的抗癌效果。机制研究表明，去除LPS可避免由LPS引起的免疫细胞死亡，并减少野生型OMV对肿瘤免疫细胞浸润的负面影响。我们通过掺入IL-2变体蛋白（Neo-2/15）进一步展示了无LPS的OMV的功能。这种功能化增强了OMV抑制肿瘤生长和促进淋巴细胞浸润到肿瘤微环境中的能力。本研究提出了一种具有改善的转化前景的安全且可功能化的OMV。
