Proteomic and metabolomic alterations in follicular fluid associated with embryological outcomes in women with isolated ANA-positive infertility: a prospective multi-omics cohort study.
- 1Department of Reproductive Medicine, Women's Hospital of Nanjing Medical University, Nanjing Women and Children's Healthcare Hospital, 123 Tianfeixiang, Mochou Road, Qinhuai District, Nanjing, JiangSu, China.
- 2Department of Reproductive Medicine, Women's Hospital of Nanjing Medical University, Nanjing Women and Children's Healthcare Hospital, 123 Tianfeixiang, Mochou Road, Qinhuai District, Nanjing, JiangSu, China. [email protected].
- 3Department of Reproductive Medicine, Women's Hospital of Nanjing Medical University, Nanjing Women and Children's Healthcare Hospital, 123 Tianfeixiang, Mochou Road, Qinhuai District, Nanjing, JiangSu, China. [email protected].
Abstract
Background
Autoimmune abnormalities are increasingly recognized as contributing factors to unexplained infertility. Among these, antinuclear antibodies (ANAs) are the most frequently detected autoantibodies and have been associated with adverse embryologic outcomes. However, the mechanisms by which isolated ANA positivity (defined as ANA positivity in the absence of anti-extractable nuclear antigen [anti-ENA] antibodies) may influence oocyte developmental competence and early embryonic development remain poorly understood. This study aimed to characterize follicular fluid (FF) molecular alterations associated with isolated ANA positivity using an integrative multi-omics approach.Methods
In this prospective cohort study, 70 infertile women undergoing their first assisted reproductive technology (ART) cycle were enrolled, including 32 women with isolated ANA positivity (serum ANA ≥ 1:80 and anti-ENA negative) and 38 ANA-negative controls. FF samples collected on the day of oocyte retrieval were subjected to integrated metabolomic and proteomic analyses. Differential expression and functional enrichment analyses were performed, followed by correlation and network analyses to explore associations between FF molecular profiles and oocyte as well as embryo developmental competence. Key metabolites were further evaluated using receiver operating characteristic (ROC) analyses to characterize their exploratory discriminatory performance.Results
Women with isolated ANA positivity showed significantly poorer embryo developmental outcomes than ANA-negative controls, including lower IVF/ICSI fertilization rates and reduced numbers of high-quality embryos and blastocysts (all P < 0.05). After adjustment for potential confounders, multivariate linear regression analysis suggested that isolated serum ANA positivity was independently associated with impaired early embryonic development (P < 0.05). Integrated multi-omics analyses revealed significant dysregulation of pathways related to steroid hormone biosynthesis, immune-related pathways including complement and coagulation cascades, amino acid metabolism, and glycerophospholipid metabolism. Among the identified molecular features, three metabolites-L-phenylalanine, SM(d18:1/16:0), and hydrocortisone-were consistently associated with embryo developmental potential and showed stable correlations independent of ANA titers. A combined preliminary metabolite panel was identified that discriminates ANA-positive from ANA-negative women with excellent discriminatory performance (AUC = 0.968). In exploratory analyses of dichotomized oocyte- and early embryo-related outcomes, the panel showed moderate discriminatory performance (AUCs = 0.719-0.771).Conclusions
Isolated ANA positivity is associated with metabolic reprogramming in the follicular microenvironment and impaired early embryonic development. The identified exploratory metabolite signatures provide insight into immune-metabolic alterations that may underlie ANA-associated infertility and may inform future prognostic model development. Their associations appeared consistent across the ANA-titer subgroups examined. However, the panel remains preliminary and at the discovery stage and requires validation in independent cohorts before its clinical relevance or utility can be determined.
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BibTeX
@article{ding2026proteomic,
title = {{Proteomic and metabolomic alterations in follicular fluid associated with embryological outcomes in women with isolated ANA-positive infertility: a prospective multi-omics cohort study.}},
author = {Kai Ding and Wei Jiang and Yidan Chen and Shanren Cao and Xin Li and Jiajia Yao and Yi Wei and Junqiang Zhang and Xiufeng Ling and Chun Zhao},
journal = {Reproductive biology and endocrinology : RB\&E},
year = {2026},
month = aug,
doi = {10.1186/s12958-026-01599-w},
url = {https://europepmc.org/article/MED/42958402},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Proteomic and metabolomic alterations in follicular fluid associated with embryological outcomes in women with isolated ANA-positive infertility: a prospective multi-omics cohort study.
AU - Kai Ding
AU - Wei Jiang
AU - Yidan Chen
AU - Shanren Cao
AU - Xin Li
AU - Jiajia Yao
AU - Yi Wei
AU - Junqiang Zhang
AU - Xiufeng Ling
AU - Chun Zhao
PY - 2026
DA - 2026/08/01
JO - Reproductive biology and endocrinology : RB&E
DO - 10.1186/s12958-026-01599-w
UR - https://europepmc.org/article/MED/42958402
AB - Background Autoimmune abnormalities are increasingly recognized as contributing factors to unexplained infertility. Among these, antinuclear antibodies (ANAs) are the most frequently detected autoantibodies and have been associated with adverse embryologic outcomes. However, the mechanisms by which isolated ANA positivity (defined as ANA positivity in the absence of anti-extractable nuclear antigen [anti-ENA] antibodies) may influence oocyte developmental competence and early embryonic development remain poorly understood. This study aimed to characterize follicular fluid (FF) molecular alterations associated with isolated ANA positivity using an integrative multi-omics approach.Methods In this prospective cohort study, 70 infertile women undergoing their first assisted reproductive technology (ART) cycle were enrolled, including 32 women with isolated ANA positivity (serum ANA ≥ 1:80 and anti-ENA negative) and 38 ANA-negative controls. FF samples collected on the day of oocyte retrieval were subjected to integrated metabolomic and proteomic analyses. Differential expression and functional enrichment analyses were performed, followed by correlation and network analyses to explore associations between FF molecular profiles and oocyte as well as embryo developmental competence. Key metabolites were further evaluated using receiver operating characteristic (ROC) analyses to characterize their exploratory discriminatory performance.Results Women with isolated ANA positivity showed significantly poorer embryo developmental outcomes than ANA-negative controls, including lower IVF/ICSI fertilization rates and reduced numbers of high-quality embryos and blastocysts (all P < 0.05). After adjustment for potential confounders, multivariate linear regression analysis suggested that isolated serum ANA positivity was independently associated with impaired early embryonic development (P < 0.05). Integrated multi-omics analyses revealed significant dysregulation of pathways related to steroid hormone biosynthesis, immune-related pathways including complement and coagulation cascades, amino acid metabolism, and glycerophospholipid metabolism. Among the identified molecular features, three metabolites-L-phenylalanine, SM(d18:1/16:0), and hydrocortisone-were consistently associated with embryo developmental potential and showed stable correlations independent of ANA titers. A combined preliminary metabolite panel was identified that discriminates ANA-positive from ANA-negative women with excellent discriminatory performance (AUC = 0.968). In exploratory analyses of dichotomized oocyte- and early embryo-related outcomes, the panel showed moderate discriminatory performance (AUCs = 0.719-0.771).Conclusions Isolated ANA positivity is associated with metabolic reprogramming in the follicular microenvironment and impaired early embryonic development. The identified exploratory metabolite signatures provide insight into immune-metabolic alterations that may underlie ANA-associated infertility and may inform future prognostic model development. Their associations appeared consistent across the ANA-titer subgroups examined. However, the panel remains preliminary and at the discovery stage and requires validation in independent cohorts before its clinical relevance or utility can be determined.
SN - 1477-7827
ER -