Cell-free DNA in Human Follicular Microenvironment: New Prognostic Biomarker to Predict in vitro Fertilization Outcomes

Abstract

Cell-free DNA (cfDNA) fragments, detected in blood and in other biological fluids, are released from apoptotic and/or necrotic cells. CfDNA is currently used as biomarker for the detection of many diseases such as some cancers and gynecological and obstetrics disorders. In this study, we investigated if cfDNA levels in follicular fluid (FF) samples from in vitro fertilization (IVF) patients, could be related to their ovarian reserve status, controlled ovarian stimulation (COS) protocols and IVF outcomes. Therefore, 117 FF samples were collected from women (n = 117) undergoing IVF/Intra-cytoplasmic sperm injection (ICSI) procedure and cfDNA concentration was quantified by ALU-quantitative PCR. We found that cfDNA level was significantly higher in FF samples from patients with ovarian reserve disorders (low functional ovarian reserve or polycystic ovary syndrome) than from patients with normal ovarian reserve (2.7 ± 2.7 ng/μl versus 1.7 ± 2.3 ng/μl, respectively, p = 0.03). Likewise, FF cfDNA levels were significant more elevated in women who received long ovarian stimulation (> 10 days) or high total dose of gonadotropins (≥ 3000 IU/l) than in women who received short stimulation duration (7–10 days) or total dose of gonadotropins < 3000 IU/l (2.4 ± 2.8 ng/μl versus 1.5 ± 1.9 ng/μl, p = 0.008; 2.2 ± 2.3 ng/μl versus 1.5 ± 2.1 ng/μl, p = 0.01, respectively). Finally, FF cfDNA level was an independent and significant predictive factor for pregnancy outcome (adjusted odds ratio = 0.69 [0.5; 0.96], p = 0.03). In multivariate analysis, the Receiving Operator Curve (ROC) analysis showed that the performance of FF cfDNA in predicting clinical pregnancy reached 0.73 [0.66–0.87] with 88% specificity and 60% sensitivity. CfDNA might constitute a promising biomarker of follicular micro-environment quality which could be used to predict IVF prognosis and to enhance female infertility management.

Mitochondrial dysfunction in cumulus-oocyte complexes increases cell-free mitochondrial DNA

“…cfDNA concentration in human FF is known to decrease with the development of the follicle; its concentration rangs from 1 to 3 ng/μl in the antral follicles (8−12 mm in diameter) [ 9 , 15 , 16 ]. Consistent with this, the present study showed that the cfDNA content in FF decreased with follicle development, and the concentration in FF derived from S-, M-, and L-FF was 16.4 ± 0.5, 10.5 ± 0.7, and 2.2 ± 0.2 ng/μl, respectively ( Fig.…”

“…A pioneering study has shown that the amount of cfDNA in FF was related to the apoptosis of granulosa cells and that addition of cfDNA to the culture medium induced the latter [ 10 ]. On the other hand, accumulating evidence suggests that the amount of cfDNA reflects the ovarian reserve and possible outcome of in vitro fertilization [ 15 , 16 ]. In the present study, however, the phenomenon by which cfDNA (present in FF or culture medium) reflects oocyte ability and follicle condition has not been elucidated.…”

Сell-free D N A in follicular fluid of women with different parameters of ovarian function

“…В настоящем исследовании продемонстрировано, что содержание свДНК в ФЖ женщин со стимулированной овуляцией значимо выше, чем в естественных циклах. Полученные результаты согласуются с данными других авторов, которые выявили наличие свДНК в ФЖ женщин со стимулированной овуляцией [14,[19][20][21] и показали прямую взаимосвязь между концентрацией свДНК и дозой гонадотропина [14]. Данные о наличии свДНК в ФЖ женщин в естественных циклах ранее в литературе не освещались и, учитывая иммуномодулирующие свойства ДНК [5], могут послужить основой для исследования новых аспектов иммунно-ýндокринных взаимодействий в регуляции фолликулогенеза и созревания ооцитов.…”

“…Так, S. Traver и соавт. выявили повышенный уровень свДНК в группе женщин с СПКЯ [14]. В настоящем исследовании продемонстрировано, что наличие большого числа АФ (>14) у женщин с мультифолликулярными яичниками также ассоциируется с высокой концентрацией свДНК.…”

“…Среди различных биомаркеров патологических процессов свободная внеклеточная ДНК (свДНК) в последнее время привлекает большое внимание [1]. Несвязанная с клетками ДНК выявляется в различных биологических жидкостях организма [2][3][4], ее концентрация возрастает при целом ряде заболеваний (онкологические, аутоиммунные, посттравматический синдром) [5][6][7][8][9], включая акушерско-гинекологическую патологию [10], что обусловливает интерес к исследованию диагностической и прогностической значимости свДНК [11][12][13][14].…”