在育龄妇女中,阴道微生物组通常由一种或几种乳杆菌组成,包括crispatus,乳酸菌,Paragasseri乳杆菌,木乳杆菌,和卷曲乳杆菌,与最佳的宫颈阴道健康有关;然而,关于其他乳杆菌代谢如何促进宫颈阴道健康,仍然未知。我们假设每种乳杆菌的代谢物不同,并且对健康和体内平衡有独特的贡献。为了解决这个假设,我们利用人类三维(3D)宫颈上皮细胞模型与基因组学分析和非靶向代谢组学相结合,以确定研究较少的阴道乳杆菌的代谢贡献。iners,L.paragasseri,还有L.mulieris.我们的研究验证了阴道乳杆菌表现出密切的系统发育关系。来自公开可用菌株和我们研究中使用的菌株的基因组发现表明,内乳杆菌在代谢上与其他乳酸杆菌不同,可能是由于基因组大小减小。乳杆菌和模拟对照基于全局代谢谱是可区分的。我们鉴定了个体乳杆菌和模拟对照之间95个显著改变的代谢物(P<0.05)。与氨基酸代谢相关的代谢物在乳杆菌中共享。具有潜在抗微生物特性的N-乙酰化氨基酸以物种特异性方式显着升高。L.paragasseri和L.iners共享芳香,但不是碳水化合物衍生的,具有潜在抗菌特性的乳酸代谢物可能有助于宫颈阴道环境的稳态。此外,L.iners独特地改变了脂质代谢,这可能是适应宫颈阴道生态位的标志。总的来说,这些发现进一步阐明了三种关键的阴道乳杆菌在妇科健康中的代谢贡献.重要性乳杆菌属物种通过产生乳酸和其他抗微生物化合物而有助于宫颈阴道健康。然而,关于较少研究但常见的阴道乳杆菌的代谢潜力,仍然未知。这里,我们使用非靶向代谢组学结合我们的3D宫颈上皮细胞模型来识别阴道乳杆菌种类之间的代谢差异(乳杆菌,Paragasseri乳杆菌,和mulieris乳杆菌)以及这些差异如何与维持宫颈上皮的稳态有关。人类3D细胞模型是研究宿主细菌相互作用和减少临床研究中固有的混杂因素的重要工具。因此,这些独特的模型使我们能够破译假定的乳杆菌机制,这些机制有助于它们在健康或疾病中的作用。代谢分析揭示了每个乳杆菌属物种的不同概况,但也共享与抗菌活性相关的代谢贡献:氨基酸代谢,N-乙酰化氨基酸,和芳香乳酸。这些模式在临床研究中提供了与健康相关的代谢物的验证,并提供了新的靶标。包括免疫调节和抗菌代谢物,用于后生物疗法。
In reproductive-age women, the vaginal microbiome is typically dominated by one or a few Lactobacillus species, including Lactobacillus crispatus, Lactobacillus iners, Lactobacillus paragasseri, Lactobacillus mulieris, and Lactobaccillus crispatus, has been associated with optimal cervicovaginal health; however, much is still unknown about how other lactobacilli metabolically contribute to cervicovaginal health. We hypothesized that metabolites of each Lactobacillus species differ and uniquely contribute to health and homeostasis. To address this hypothesis, we utilized a human three-dimensional (3D) cervical epithelial cell model in conjunction with genomics analyses and untargeted metabolomics to determine the metabolic contributions of less-studied vaginal lactobacilli-L. iners, L. paragasseri, and L. mulieris. Our study validated that vaginal lactobacilli exhibit a close phylogenetic relationship. Genomic findings from publicly available strains and those used in our study indicated that L. iners is metabolically distinct from other species of lactobacilli, likely due to a reduced genome size. Lactobacilli and mock controls were distinguishable based on global metabolic profiles. We identified 95 significantly altered metabolites (P < 0.05) between individual lactobacilli and mock controls. Metabolites related to amino acid metabolism were shared among the lactobacilli. N-Acetylated amino acids with potential antimicrobial properties were significantly elevated in a species-specific manner. L. paragasseri and L. iners shared aromatic, but not carbohydrate-derived, lactic acid metabolites with potential antimicrobial properties that may contribute to homeostasis of the cervicovaginal environment. Additionally, L. iners uniquely altered lipid metabolism, which may be a sign of adaptation to the cervicovaginal niche. Overall, these findings further elucidate the metabolic contributions of three key vaginal Lactobacillus species in gynecological health. IMPORTANCE Lactobacillus species contribute to cervicovaginal health by their production of lactic acid and other antimicrobial compounds. Yet, much is still unknown regarding the metabolic potential of lesser-studied but common vaginal lactobacilli. Here, we used untargeted metabolomics coupled with our 3D cervical epithelial cell model to identify metabolic differences among vaginal Lactobacillus species (Lactobacillus iners, Lactobacillus paragasseri, and Lactobacillus mulieris) and how those differences related to maintaining homeostasis of the cervical epithelium. Human 3D cell models are essential tools for studying host-bacteria interactions and reducing confounding factors inherent in clinical studies. Therefore, these unique models allowed us to decipher the putative lactobacilli mechanisms that contribute to their roles in health or disease. Metabolic analyses revealed distinct profiles of each Lactobacillus species but also shared metabolic contributions associated with antimicrobial activity: amino acid metabolism, N-acetylated amino acids, and aromatic lactic acids. These patterns provided validation of metabolites associated with health in clinical studies and provided novel targets, including immunomodulatory and antimicrobial metabolites, for postbiotic therapies.