Exposure of human fallopian tube epithelium to elevated testosterone results in alteration of cilia gene expression and beating

Abstract

STUDY QUESTION How does exposure to a testosterone rich environment affect the function and gene expression of human fallopian tube epithelium (hFTE)? SUMMARY ANSWER Elevated testosterone level alters several gene transcripts that regulate cilia expression and negatively impacts the rate of cilia beating. WHAT IS KNOWN ALREADY The presence of estrogen in the follicular phase of the menstrual cycle increases the human fallopian tube ciliary beating frequency. The luteal phase, triggered by ovulation and increasing progesterone, is marked by a decrease in ciliary beating. Women with polycystic ovarian syndrome (PCOS) may have twice the serum level of testosterone than ovulatory women. To date, the effect of elevated androgens on the function of the human fallopian tube is not well-understood. We chose to examine the impact of elevated testosterone on hFTE. STUDY DESIGN, SIZE, DURATION A prospective basic science study of human fallopian tube specimens from reproductive-aged women undergoing benign gynecologic surgery was performed. Fallopian tube removal at a large US academic center was collected and provided to us to continue with epithelium isolation and culturing. A total of 12 patients were analyzed in the study. PARTICIPANTS/MATERIALS, SETTING, METHODS Fallopian tube epithelium was isolated and exposed to two different conditions: normal with low testosterone concentration of 0.8 nM and PCOS-like, with high testosterone concentration of 2 nM. The study was conducted in both static and dynamic conditions in microfluidic devices for a total of 14 days, after which the tissue was collected for processing including RNA extraction, quantitative PCR and immunohistochemistry. After the first 7 days of each experiment, a sample of tissue from each condition was imaged to quantify cilia beating frequency. MAIN RESULTS AND THE ROLE OF CHANCE hFTE exposed to the 2 nM testosterone displayed slower cilia beating, inhibited estrogen signaling and decreased expression of the ciliary marker FOXJ1 when compared to stimulation with 0.8 nM testosterone. LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION The in vivo response to elevated testosterone may differ from in vitro studies. RNA amount was limited from tissue cultured in the microfluidic devices as compared to static culture. WIDER IMPLICATIONS OF THE FINDINGS Understanding elevated testosterone in tubal function may explain an additional contribution to subfertility in women with PCOS and other hyper-androgen disorders, aside from oligo-ovulation. Furthermore, this adds to the body of literature of fallopian tube function using a microfluidic device. STUDY FUNDING/COMPETING INTEREST(S) NIH grants: UH3 ES029073 and R01 CA240301. There are no competing interests.

Fallopian Tube-Derived Tumor Cells Induce Testosterone Secretion from the Ovary, Increasing Epithelial Proliferation and Invasion

“…Only the fimbriated ends of the fallopian tube that are in proximity of the ovary display serous tubal intraepithelial carcinoma in situ, a lesion thought to give rise to high grade serous cancer. Endocrine signals from the ovary also play a role in physiological changes in the FTE to support functions such as gamete and embryo transport, tubal secretions, and sperm capacitation [ 40 , 41 , 42 , 43 ]. Previously, we have shown that tumorigenic fallopian tube cells can differentially modify the metabolome of the ovary as compared to nontumorigenic controls.…”

Isolation of Fallopian Tube Epithelium for Assessment of Cilia Beating Frequency (CBF)

“…Estrogen (1 nM) increases CBF, whereas progesterone (10 nM) and testosterone (2 nM) reduce CBF. Basal CBF observed with 0.1 nM estrogen stimulation is about 20 Hz [13,14].…”

“…Estrogen stimulates ciliogenesis and increases cilia beating frequency while high levels of progesterone cause deciliation and decrease beating frequency [12,13]. Elevated testosterone also regulates cilia gene expression and motion in human fallopian tube epithelium [14]. Human fallopian secretory epithelial cells are a source of origin of highgrade serous ovarian cancer, and the outgrowth of the secretory population, as compared to the ciliated cells, is thought to be an early event in tumorigenesis [15,16].…”

Top‐quality embryo transfer is associated with lower odds of ectopic pregnancy

“…In the Japanese analysis, 23 EP rate after ovarian stimulation was compared with the rate after spontaneous cycle IVF/ICSI and was found to increase the odds of EP by 3.5–5.2. Recently, several studies have shown that Fallopian tube epithelium is sensitive to circulating hormonal levels through changes in the expression of steroid receptors 24 that affect Fallopian epithelial cilia movements 25,26 . In light of these findings, Jwa and colleagues have speculated that the higher hormonal levels associated with ovarian stimulation for IVF/ICSI are disruptive for the function of the Fallopian tube 23 .…”