Medici Giorgio, Candini Giulia, Mottolese Nicola, Uguagliati Beatrice, Trebbi Federica, Loi Manuela, Bove Angelica Marina, Stojanov Spase, Esposito Erika, Vitagliano Rosalba, D'Amico Federica, Turroni Silvia, Fiori Jessica, Berlec Aleš, Trazzi Stefania, Ciani Elisabetta
Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, Italy.
Department of Biotechnology, Jozef Stefan Institute, Ljubljana, Slovenia.
J Biol Eng. 2025 Jul 17;19(1):65. doi: 10.1186/s13036-025-00538-4.
Protein-based therapeutics have garnered increasing attention across various medical fields for their substantial benefits to human health. Existing strategies for intracellular protein delivery, such as cell-penetrating peptide (CPP)-based approaches, including the transactivator of transcription (TAT) peptide, have shown promising results but also present limitations, particularly due to the need for recombinant protein manufacturing and daily invasive administration. To overcome such hindrances and develop delivery tools that are able to foster the production of the protein directly inside the body of patients, we engineered the lactic acid bacterium, Lactococcus lactis, to express and secrete TATκ-GFP protein. After oral administration of the recombinant L. lactis in mice, we found the presence of the TATκ-GFP protein not only in the intestinal wall but also in the mouse liver, heart, and brain. This provides the first evidence that a recombinant TATκ-fused protein, secreted by L. lactis in the mouse host gut, by virtue of the presence of the TATκ peptide, can efficiently pass from the intestinal wall to other organs. With future research and development, this technology based on lactic acid bacteria (LAB) as a platform for the production and delivery of therapeutic proteins could offer significant advancements in the field of therapeutic protein delivery.
基于蛋白质的疗法因其对人类健康的巨大益处而在各个医学领域受到越来越多的关注。现有的细胞内蛋白质递送策略，如基于细胞穿透肽（CPP）的方法，包括转录激活因子（TAT）肽，已显示出有前景的结果，但也存在局限性，特别是由于需要重组蛋白制造和每日侵入性给药。为了克服这些障碍并开发能够促进在患者体内直接产生蛋白质的递送工具，我们对乳酸乳球菌进行了工程改造，使其表达并分泌TATκ-GFP蛋白。在小鼠口服重组乳酸乳球菌后，我们发现TATκ-GFP蛋白不仅存在于肠壁中，还存在于小鼠的肝脏、心脏和大脑中。这提供了首个证据，即乳酸乳球菌在小鼠宿主肠道中分泌的重组TATκ融合蛋白，凭借TATκ肽的存在，能够有效地从肠壁传递到其他器官。随着未来的研发，这种基于乳酸菌（LAB）作为治疗性蛋白质生产和递送平台的技术，有望在治疗性蛋白质递送领域取得重大进展。
