mSystems

One-year restoration of vaginal health: synergistic dynamics of microbiome and metabolome following the elimination of high-grade cervical intraepithelial neoplasia

ABSTRACT Therapeutic elimination of high-grade cervical intraepithelial neoplasia (CIN) is widely implemented for cervical cancer prevention. Despite the demonstrated dysbiosis of vaginal microenvironment in high-grade CIN, its post-therapy restorations remain to be poorly understood, especially in functional aspects. This study aimed to characterize temporal changes in both vaginal microbiota (VM) and metabolome (VMeta) following therapeutic elimination of high-grade CIN. We conducted a longitudinal study of 32 HPV-positive women with high-grade CIN who underwent therapeutic procedures. Vaginal swabs were collected at baseline (pre-therapy) and at 6- and 12-month follow-up visits for integrated VM and VMeta analysis. We observed a gradual restoration of Lactobacillus crispatus levels from baseline to 12 months ( P < 0.05). Concurrently, we detected significant decreases in dysbiosis-associated bacteria, including Prevotella bivia , Ureaplasma parvum , and Peptoniphilus sp. 6 months post-therapy compared to the baseline. VMeta analysis revealed distinct metabolic shifts across the follow-up periods. The early post-therapy phase (baseline to 6 months) was characterized by enrichment of glycerophospholipids and depletion of nucleotide metabolites, while the later phase (6–12 months) showed increases in flavonoids, lysophospholipids, bioactive amides, and amino acid metabolism. Integration of correlation and dynamic Bayesian network analysis indicated potential regulatory relationships and time-lag effects involving HPV infection, L. crispatus , Bifidobacterium sp., Streptococcus anginosus , Megasphaera sp., U. parvum, and those metabolites. This study enhances our understanding of a sequential restoration process post-therapy in the vaginal microenvironment. IMPORTANCE Therapeutic elimination of high-grade CIN is routine, yet functional recovery of the vaginal ecosystem is poorly defined. In a 12-month longitudinal multi-omics study of 32 women, we show stepwise restoration: progressive L. crispatus dominance with sustained decreases in dysbiosis-associated taxa ( P. bivia, U. parvum, Peptoniphilus ). Metabolically, an early rise in glycerophospholipids and fall in nucleotide metabolites is followed by later enrichment of flavonoids, lysophospholipids, bioactive amides, and amino acid derivatives. Correlation and dynamic Bayesian network analyses reveal putative regulatory links, time-lag effects, and downstream impacts of HPV clearance. These findings deliver a functional roadmap of post-therapy recovery, nominate measurable microbial–metabolite milestones and candidate biomarkers for monitoring, and suggest targets for adjunct interventions to accelerate re-establishment of protective states. This work informs precision follow-up in cervical cancer prevention programs.

Lactobacillus crispatus -derived nCEV vesicles promote cutaneous wound healing and inhibit HPV16 infection

ABSTRACT Cervical cancer (CC) is a common gynecological malignancy, primarily associated with persistent infection by high-risk human papillomavirus (HPV). The vaginal microbiota, dominated by Lactobacillus species, plays a critical role in maintaining a healthy vaginocervical microenvironment. However, the influence of Lactobacillus on the pathogenesis and treatment of CC remains underexplored. Here, we report that the optimized 60–80 nm size dominant of extracellular vesicles from various Lactobacillus species (nLEVs) significantly affects cutaneous wound healing and HPV infection. Specifically, nCEVs from Lactobacillus crispatus demonstrate superior efficacy compared to nIEVs from Lactobacillus iners in promoting cell migration, angiogenesis, and wound healing via macrophage M2 polarization and blocking intravaginal HPV16 infection. Moreover, metabolic profiling revealed that D-lactate may be key to the functions of nCEVs. Altogether, our findings uncover a novel nCEV/D-lactate-mediated mechanism that promotes homeostasis and offers potential new approaches for the prevention and treatment of CC. IMPORTANCE This study identifies Lactobacillus crispatus -derived vesicles (nCEVs) as crucial for cervicovaginal homeostasis, functioning through promoting wound healing via macrophage polarization and blocking HPV infection, and delivering D-lactate—a key bioactive component. These insights advance microbiome-based female healthcare interventions.

Gut-derived metabolite 3-methylxanthine enhances cisplatin-induced apoptosis via dopamine receptor D1 in a mouse model of ovarian cancer

ABSTRACT Platinum-based chemotherapy failure represents a significant challenge in the management of ovarian cancer (OC) and contributes to disease recurrence and poor prognosis. Recent studies have shed light on the involvement of the gut microbiota in modulating anticancer treatments. However, the precise underlying mechanisms, by which gut microbiota regulates the response to platinum-based therapy, remain unclear. Here, we investigated the role of gut microbiota on the anticancer response of cisplatin and its underlying mechanisms. Our results demonstrate a substantial improvement in the anticancer efficacy of cisplatin following antibiotic-induced perturbation of the gut microbiota in OC-bearing mice. 16S rRNA sequencing showed a pronounced alteration in the composition of the gut microbiome in the cecum contents following exposure to cisplatin. Through metabolomic analysis, we identified distinct metabolic profiles in the antibiotic-treated group, with a notable enrichment of the gut-derived metabolite 3-methylxanthine in antibiotic-treated mice. Next, we employed a strategy combining transcriptome analysis and chemical–protein interaction network databases. We identified metabolites that shared structural similarity with 3-methylxanthine, which interacted with genes enriched in cancer-related pathways. It is identified that 3-methylxanthinesignificantly enhances the effectiveness of cisplatin by promoting apoptosis both in vivo and in vitro . Importantly, through integrative multiomics analyses, we elucidated the mechanistic basis of this enhanced apoptosis, revealing a dopamine receptor D1-dependent pathway mediated by 3-methylxanthine. This study elucidated the mechanism by which gut-derived metabolite 3-methylxanthine mediated cisplatin-induced apoptosis. Our findings highlight the potential translational significance of 3-methylxanthine as a promising adjuvant in conjunction with cisplatin, aiming to improve treatment outcomes for OC patients. IMPORTANCE The precise correlation between the gut microbiota and the anticancer effect of cisplatin in OC remains inadequately understood. Our investigation has revealed that manipulation of the gut microbiota via the administration of antibiotics amplifies the efficacy of cisplatin through the facilitation of apoptosis in OC-bearing mice. Metabolomic analysis has demonstrated that the cecum content from antibiotic-treated mice exhibits an increase in the levels of 3-methylxanthine, which has been shown to potentially enhance the therapeutic effectiveness of cisplatin by an integrated multiomic analysis. This enhancement appears to be attributable to the promotion of cisplatin-induced apoptosis, with 3-methylxanthine potentially exerting its influence via the dopamine receptor D1-dependent pathway. These findings significantly contribute to our comprehension of the impact of the gut microbiota on the anticancer therapy in OC. Notably, the involvement of 3-methylxanthine suggests its prospective utility as a supplementary component for augmenting treatment outcomes in patients afflicted with ovarian cancer.

Microbiome and metabolomic changes associated with HPV clearance in women undergoing local excisional treatment for cervical intraepithelial neoplasia

ABSTRACT Cervical intraepithelial neoplasia (CIN) is a common gynecological condition often associated with persistent human papillomavirus (HPV) infections. Although the loop electrosurgical excision procedure (LEEP) is effective in removing lesions, some patients remain HPV positive post-treatment. In this prospective study, we enrolled reproductive-age women diagnosed with HPV-related CIN and employed a multi-omics analysis of cervicovaginal secretion and cervical tissue microbiomes, alongside non-targeted and targeted metabolomic assessments. We aim to explore the role of the cervicovaginal and intratissue microbiota and associated metabolites on HPV clearance following LEEP. We observed significant shifts in bacterial diversity and composition in both cervicovaginal secretion and cervical tissue samples. Notably, distinct bacterial species, such as Lactobacillus and certain anaerobes (e.g., Prevotella bivia ), were correlated with HPV clearance post-LEEP. Metabolomic profiling revealed that the HPV-cleared group exhibited elevated acetic acid levels and significant alterations in glycerophospholipid metabolism, suggesting a potential role in promoting HPV clearance. Correlation analyses demonstrated significant associations between altered bacteria and metabolites with HPV status, with models incorporating both achieving high predictive accuracy. Overall, our findings highlight the importance of the cervicovaginal and intratissue microbiomes and metabolites in facilitating HPV clearance, suggesting potential therapeutic targets for patients with CIN. IMPORTANCE The clearance of human papillomavirus (HPV) after local excisional treatment for cervical intraepithelial neoplasia is crucial for patient health. This study reveals significant alterations in the cervicovaginal secretion and cervical tissue microbiomes, alongside metabolomic changes, which are associated with HPV clearance. Through a comprehensive multi-omics approach, we identified specific bacterial species and metabolic changes that correlate with successful HPV clearance post-loop electrosurgical excision procedure. Notably, the presence of beneficial Lactobacillus species and elevated levels of acetic acid linked to glycerophospholipid metabolism emerged as potential biomarkers for HPV status, suggesting that these factors play a pivotal role in improving treatment outcomes. These findings highlight the potential for microbiome-targeted therapies to enhance HPV clearance and provide insights into the microbial and metabolic mechanisms involved in cervical health.

Cervicovaginal DNA Virome Alterations Are Associated with Genital Inflammation and Microbiota Composition

HPV infection is an established risk factor for cervical cancer. However, more broadly, the role of the cervicovaginal virome in cervical cancer progression is not well understood.