The reproductive tract microbiome in women with polycystic ovary syndrome and across different menstrual cycle phases.

Study Question: Do polycystic ovary syndrome (PCOS), menstrual cycle phases, and ovulatory status affect reproductive tract (RT) microbiome profiles?

Summary Answer: We identified microbial features associated with menstrual cycle phases in the upper and lower RT microbiome, but only two specific differences in the upper RT according to PCOS status.

What Is Known Already: The vaginal and uterine microbiome profiles vary throughout the menstrual cycle. Studies have reported alterations in the vaginal microbiome among women diagnosed with PCOS.

Study Design, Size, Duration: This prospective case-control study included a cohort of 37 healthy control women and 52 women diagnosed with PCOS. Microbiome samples were collected from the vagina as vaginal swabs (VS) and from the uterus as endometrial flushing (EF) aspirate samples, and compared according to PCOS diagnosis, the menstrual cycle phases, and ovulatory status, at Oulu University Hospital (Oulu, Finland) from January 2017 to March 2020.

Participants/materials, Setting, Methods: A total of 83 VS samples and 80 EF samples were collected. Age and body mass index (BMI) were matched between women with and without PCOS. Clinical characteristics were assessed using blood samples collected between cycle days 2 and 8, and microbial DNA was sequenced on the Ion Torrent platform. Microbial alpha diversity (i.e. the observed number of unique genera and Shannon diversity index) was analysed across sample types, PCOS diagnosis and menstrual cycle phases. Linear mixed-effects models were utilised to identify microbial features in relation to PCOS and the menstrual cycle phases. Associations between the beta diversity of the RT microbiome and PCOS- and cycle-related clinical features were calculated using PERMANOVA.

Main Results And The Role Of Chance: Microbial alpha diversity showed no difference with PCOS (VS: Pobserved feature = 0.836, Pshannon = 0.998; EF: Pobserved feature = 0.366, Pshannon = 0.185), but varied with menstrual cycle phases (VS: Pobserved feature = 0.001, Pshannon = 0.882; EF: Pobserved feature = 0.026, Pshannon = 0.048). No difference was observed in beta diversity based on either PCOS or the menstrual cycle phases (VS: PPCOS = 0.280, Pcycle = 0.115; EF: PPCOS = 0.234, Pcycle = 0.088). In the endometrial flushing samples, we identified two novel microbial features, characterised by the ratio of differential abundance of two genera, associated with PCOS (FDR ≤ 0.1) and 13 novel features associated with the menstrual cycle phases (FDR ≤ 0.1).

Limitations, Reasons For Caution: Although this was the first study to simultaneously analyse, the lower and upper RT microbiome in women with and without PCOS, the limited sample size of anovulatory cases may hinder the detection of differences related to PCOS and ovulatory status.

Wider Implications Of The Findings: The main finding suggests that PCOS and the menstrual cycle phases are associated with specific microbial features in the upper RT, indicating that the analysis of the upper RT microbiome can potentially identify biomarkers for both PCOS and menstrual cycle phases.

Study Funding/competing Interest(s): This research was funded by the Research Council of Finland (grants no. 315921, 321763, 336449), the Sigrid Jusélius Foundation, Novo Nordisk Foundation (grant no. NNF21OC0070372), and the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant (MATER, grant no. 813707). This research was also funded by the Estonian Research Council (grants no. PRG1076, PRG1414), the Horizon Europe grant (NESTOR, grant no. 101120075) of the European Commission, and EMBO Installation Grant (grant no. 3573). The funders did not participate in any processes of the study. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Trial Registration Number: N/A.