Volume 70, December 2025, 101237Author links open overlay panel, , , , Highlights•

BRCA1 mutation status influences cytokine levels in periovulatory follicular fluid.

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RANKL and CSF-1 levels are higher in follicular fluid from BRCA1 mutation carriers.

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RANKL and CSF-1 increase NFκB signaling in fallopian tube epithelial cells.

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RANKL and CSF-1 increase levels of interferon-stimulated genes.

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CSF-1 and RANKL effects in fallopian tube epithelial cells require NFκB signaling.

Abstract

Pathogenic germline mutations in BRCA1 predispose individuals to high-grade serous tubo-ovarian cancer (HGSTOC), which originates in the fallopian tube epithelium (FTE). Identified non-genetic risk factors are consistent with a potential role for repetitive exposure of FTE cells to follicular fluid during ovulation in the development of this disease. We previously showed that BRCA1 deficiency in non-malignant FTE cells associates with increased proinflammatory NFκB signaling, which could contribute to the development of HGSTOC. Additionally, exposure of BRCA1 mutated primary FTE cells to periovulatory follicular fluid resulted in further increased levels of proinflammatory gene transcripts as compared to cells isolated from control patients. In this study, we compared cytokine levels in periovulatory follicular fluid collected from BRCA1 mutation carriers to that of non-BRCA1 mutation carriers. Follicular fluid was collected from 59 patients diagnosed with breast cancer undergoing controlled ovarian stimulation and oocyte retrieval as part of their oncofertility treatment. Samples included 13 patients with confirmed BRCA1 mutations, 15 patients with mutations in other susceptibility genes and 31 patients confirmed as non-BRCA1 mutation carriers. Levels of 92 inflammatory proteins were measured using an antibody array with a proximity extension assay. Partial Least-Squares Discriminant Analysis indicated that samples from BRCA1 mutation carriers clustered separately from other samples, indicating BRCA1 mutation status influences cytokine levels in follicular fluid. RANKL and CSF-1 were among 7 proteins found to be statistically elevated in follicular fluid from BRCA1 mutation carriers. Treatment of an immortalized FTE cell line with RANKL and CSF-1 increased NFκB signaling and levels of proteins encoded by type I interferon-stimulated genes. These findings support further investigation exploring the potential of targeting RANKL and CSF-1 for HGSTOC prevention strategies in BRCA1 mutation carriers.

Keywords

Follicular fluid

Tubal-ovarian cancer

BRCA1 mutations

Ovulation

Cancer predisposition

RANKL

AbbreviationsACHP

2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)-3-pyridinecarbonitrile

ANOVA

analysis of variance

CCL5CC motif chemokine ligand 5CSF-1

colony stimulated factor-1

DMEM

Dulbecco's modified eagle medium

FTE

Fallopian tube epithelium

GnRH

gonatotropin-releasing hormone

hCG

human choriogonadotropin

HGSTOC

high-grade serous tubo-ovarian cancer

IRF3

IFN regulatory factor 3

IRF7

IFN regulatory factor 7

ISG15

IFN-stimulated gene 15

LC-MS

liquid chromatography-mass spectrometry

NPX

normalized protein expressions

OAS2

2′-5′-oligoadenylate synthetase 2

PVDF

polyvinylidene fluoride

PLS-DA

partial least-squares discriminant analysis

qPCR

quantitative polymerase chain reaction

RANK

receptor activator of nuclear factor kappa beta

RELA

nuclear factor NF-Kappa-B p65 subunit

SDS-PAGE

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

SEM

standard error of the mean

TBP

TATA-box binding protein

TNFα

tumor necrosis factor alpha

TRAIL

TNF-related apoptosis inducing ligand

USP18

ubiquitin specific peptidase 18

XTT

2,3-bis-(2‑methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide

© 2025 The Authors. Published by Elsevier Inc. CCBYLICENSE