Subcellular Localization Dictates Therapeutic Function - Spatially Targeted Delivery of Amuc 1100 by Engineered Lacticaseibacillus paracasei L9 Enhances Intestinal Barrier in Colitis

Sep 30, 2026

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Engineered Lacticaseibacillus paracasei L9 is used as a robust probiotic chassis to enable spatially targeted delivery of the Akkermansia muciniphila outer membrane protein Amuc_1100, addressing limits from the native host’s fastidious growth. Intracellular, secretory, and surface-display expression strategies are evaluated in LPS-stimulated macrophages and a DSS-induced colitis mouse model. Secretory and surface-displayed strains reduce pro-inflammatory cytokines, while secretory delivery additionally enhances immunomodulation and goblet differentiation. Surface-display delivery uniquely restores mucus-layer integrity and tight junction proteins, with distinct specialization between modes of action.

LanguageEnglish

CategoryHealthcare

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Pages22

Reading Time55 min

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Introduction

Inflammatory bowel disease and intestinal barrier dysfunction

Probiotic contributions to barrier integrity and Amuc_1100 rationale

Article Subcellular Localization Dictates Therapeutic Function: Spatially Targeted Delivery of Amuc_ 1100 by Engineered Lacticaseibacillus paracasei L9 Enhances Intestinal Barrier in Colitis Xinrui Dong 1, Li Lin 1, Weina Miao 1, Zhengyuan Zhai 1, Yanling Hao 2, Ming Zhang 3, Ran Wang 2,4, Shaoyang Ge 4, Hao Zhang 1, Lianzhong Ai 5,6 and Liang Zhao 1,2, * Academic Editor: Katsuhisa Inoue Received: 19 November 2025 Revised: 19 December 2025 Accepted: 28 December 2025 Published: 30 December 2025 Copyright: © 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license. 1 College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China; [s20213061014@cau.edu.cn](s20213061014@cau.edu.cn) (X.D.); [linli@cau.edu.cn](linli@cau.edu.cn) (L.L.); [s20233061205@cau.edu.cn](s20233061205@cau.edu.cn) (W.M.); [zhaizy@cau.edu.cn](zhaizy@cau.edu.cn) (Z.Z.); [zhanghaocau@cau.edu.cn](zhanghaocau@cau.edu.cn) (H.Z.) 2 Key Laboratory of Functional Dairy, Department of Nutrition and Health, China Agricultural University, Beijing 100193, China; [haoyl@cau.edu.cn](haoyl@cau.edu.cn) (Y.H.); [wangran@cau.edu.cn](wangran@cau.edu.cn) (R.W.) 3 School of Food and Health, Beijing Technology and Business University, Beijing 100048, China; [zhangming@th.btbu.edu.cn](zhangming@th.btbu.edu.cn) 4 Research Center for Probiotics, China Agricultural University, Beijing 101299, China; [geshaoyang@foxmail.com](geshaoyang@foxmail.com) 5 Shanghai Engineering Research Center of Food Microbiology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China; [ailianzhong@163.com](ailianzhong@163.com) 6 School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China * Correspondence: [lzhao@cau.edu.cn](lzhao@cau.edu.cn); Tel.: +86-10-62736344 Abstract Background/Objectives: Impaired intestinal barrier function is a hallmark of inflammatory bowel disease (IBD) . Akkermansia muciniphila and its outer membrane protein Amuc_1100 can enhance this barrier, but the clinical application of Amuc_ 1100 is limited by the fastidious growth of its native host. This study aimed to overcome this by utilizing the robust probiotic Lacticaseibacillus paracasei L9 for targeted Amuc_ 1100 delivery. Methods: We engineered Lc. paracasei L9 to express Amuc_ 1100 via intracellular (pA-L9), secretory (pUA-L9), and surface-display (pUPA-L9) strategies. Their efficacy was assessed in Lipopolysaccharide (LPS)-induced macrophages and a dextran sulfate sodium (DSS)-induced colitis mouse model, evaluating inflammation, barrier integrity, and mucosal repair. Results: The secretory (pUA-L9) and surface-display (pUPA-L9) strains most effectively suppressed pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α) in macrophages. In mice, both strains alleviated colitis and outperformed native A. muciniphila in improving disease activity. Crucially, they exhibited distinct, specialized functions: pUA-L9 acted as a systemic immunomodulator, reducing pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α), elevating anti-inflammatory mediators (IL-4 and IL-10), and promoting goblet cell differentiation; notably, the inhibitory effect of pUA-L9 on IL-6 expression was approximately 2-fold greater than that of pUPA-L9 . In contrast, pUPA-L9 excelled in local barrier repair, uniquely restoring mucus layer integrity (Muc1, Muc2, and Tff3) and reinforcing tight junctions (ZO-1, Occludin, Claudin1, Claudin3, and Claudin4) . In particular, pUPA-L9 increased Muc2 expression by approximately 3.6-fold compared with pUA-L9 . Conclusions: We demonstrate that the subcellular localization of Amuc_ 1100 within an engineered probiotic dictates its therapeutic mode of action. The complementary effects of secretory and surface-displayed Amuc_ 1100 offer a nove

FAQs

What problem does the study address about Amuc_1100 delivery?

Amuc_1100 can enhance intestinal barrier function, but its clinical application is limited by the fastidious growth of its native host. The study uses an engineered probiotic to overcome this growth constraint.

Which engineered L. paracasei L9 strategies were tested for Amuc_1100 delivery?

Three expression strategies were used: intracellular (pA-L9), secretory (pUA-L9), and surface-display (pUPA-L9).

How do the secretory and surface-display strains differ in therapeutic effects?

pUA-L9 mainly acts as a systemic immunomodulator by lowering pro-inflammatory cytokines and promoting goblet cell differentiation, whereas pUPA-L9 primarily excels in local barrier repair by restoring mucus-layer integrity and reinforcing tight junction proteins.

Sep 30, 2026

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