Semen levels of spermatid-specific thioredoxin-3 correlate with pregnancy rates in ART couples

Spermatid specific thioredoxin-3 (SPTRX3 or TXNDC8) is a testis/male germ line specific member of thioredoxin family that accumulates in the superfluous cytoplasm of defective human spermatozoa. We hypothesized that semen levels of SPTRX3 are reflective of treatment outcome in assisted reproductive therapy (ART) couples treated by in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Relationship between SPTRX3 and treatment outcome was investigated in 239 couples undergoing ART at an infertility clinic. Sperm content of SPTRX3 was evaluated by flow cytometry and epifluorescence microscopy, and correlated with clinical semen analysis parameters, and data on embryo development and pregnancy establishment. High SPTRX3 levels (>15% SPTRX3-positive spermatozoa) were found in 51% of male infertility patients (n¿=¿72), in 20% of men from couples with unexplained, idiopathic infertility (n¿=¿61) and in 14% of men from couples previously diagnosed with female-only infertility (n¿=¿85). Couples with high SPTRX3 produced fewer two-pronuclear zygotes and had a reduced pregnancy rate (19.2% pregnant with >15% SPTRX3-positive spermatozoa vs. 41.2% pregnant with 15% of SPTRX3-positive spermatozoa, a cutoff value established by ROC analysis, had their chance of fathering children by IVF or ICSI reduced by nearly two-thirds. The percentage of SPTRX3-positive spermatozoa had predictive value for pregnancy after ART. Gradient purification and sperm swim-up failed to remove all SPTRX3-positive spermatozoa from semen prepared for ART. In summary, the elevated semen content of SPTRX3 in men from ART couples coincided with reduced incidence of pregnancy by IVF or ICSI, identifying SPTRX3 as a candidate biomarker reflective of ART outcomeThis study was funded by grant number 1R21HD066333-01A1 from the National Institutes of Health, NICHD, grant number CB000414 from the Research Board of the University of Missouri, seed funding from the Food for The 21st Century Program of the University of Missouri and Undergraduate Summer Research Internship, College of Agriculture, Food and Natural Resources (CAFNR), University of Missouri.Peer Reviewe

Hight Throughput, Parallel Imaging and Biomarker Quantification of Human Spermatozoa by ImageStream Flow Cytometry

8 páginas, 2 figuras, 1 tabla.Spermatid specific thioredoxin-3 protein (SPTRX-3) accumulates in the superfluous cytoplasm of defective human spermatozoa. Novel ImageStream technology combining flow cytometry with cell imaging was used for parallel quantification and visualization of SPTRX-3 protein in defective spermatozoa of five men from infertile couples. The majority of the SPTRX-3 containing cells were overwhelmingly spermatozoa with a variety of morphological defects, detectable in the ImageStream recorded images. Quantitative parameters of relative SPTRX-3 induced fluorescence measured by ImageStream correlated closely with conventional flow cytometric measurements of the same sample set and reflected the results of clinical semen evaluation. Image Stream quantification of SPTRX-3 combines and surpasses the informative value of both conventional flow cytometry and light microscopic semen evaluation. The observed patterns of the retention of SPTRX-3 in the sperm samples from infertility patients support the view that SPTRX3 is a biomarker of male infertility.Peer reviewe

Semen levels of spermatid-specific thioredoxin-3 correlate with pregnancy rates in ART couples.

Spermatid specific thioredoxin-3 (SPTRX3 or TXNDC8) is a testis/male germ line specific member of thioredoxin family that accumulates in the superfluous cytoplasm of defective human spermatozoa. We hypothesized that semen levels of SPTRX3 are reflective of treatment outcome in assisted reproductive therapy (ART) couples treated by in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Relationship between SPTRX3 and treatment outcome was investigated in 239 couples undergoing ART at an infertility clinic. Sperm content of SPTRX3 was evaluated by flow cytometry and epifluorescence microscopy, and correlated with clinical semen analysis parameters, and data on embryo development and pregnancy establishment. High SPTRX3 levels (>15% SPTRX3-positive spermatozoa) were found in 51% of male infertility patients (n = 72), in 20% of men from couples with unexplained, idiopathic infertility (n = 61) and in 14% of men from couples previously diagnosed with female-only infertility (n = 85). Couples with high SPTRX3 produced fewer two-pronuclear zygotes and had a reduced pregnancy rate (19.2% pregnant with >15% SPTRX3-positive spermatozoa vs. 41.2% pregnant with <5% SPTRX3-positive sperm; one-sided p<0.05). The average pregnancy rate of all 239 couples was 25.1%. Live birth rate was 19.2% and lowest average SPTRX3 levels were found in couples that delivered twins. Men with >15% of SPTRX3-positive spermatozoa, a cutoff value established by ROC analysis, had their chance of fathering children by IVF or ICSI reduced by nearly two-thirds. The percentage of SPTRX3-positive spermatozoa had predictive value for pregnancy after ART. Gradient purification and sperm swim-up failed to remove all SPTRX3-positive spermatozoa from semen prepared for ART. In summary, the elevated semen content of SPTRX3 in men from ART couples coincided with reduced incidence of pregnancy by IVF or ICSI, identifying SPTRX3 as a candidate biomarker reflective of ART outcome

Real-world effectiveness of Fertistartkit® treatment in women undergoing ART in French clinical practice: a retrospective multicentre study

International audienceRESEARCH QUESTION:What is the real-world effectiveness of Fertistartkit® in women undergoing assisted reproductive technology (ART)?DESIGN:Retrospective cohort study including anonymized data of women undergoing ovarian stimulation for ART with Fertistartkit between April 2016 and November 2017 and follow-up of clinical outcomes up to February 2018. Data were collected from the electronic patient databases of 12 French ART centres. The main outcome was number of oocytes retrieved. All data were categorized according to female age (39 years).RESULTS:A total of 1006 cycles from 914 women treated with Fertistartkit were included. At the time of first ovarian stimulation in the study, women were 34.9 ± 5.0 years old, with a median body mass index of 22.7 kg/m². Couples had been infertile for more than 4 years, with all patterns of causes of infertility. Ovarian stimulation was started with a median dose of 300 IU (interquartile range [IQR]: 150-300 IU) of Fertistartkit for 10 days (IQR: 9-11 days), so a median total dose of 2700 IU (IQR: 1800-3300 IU). The mean number of oocytes retrieved per cycle was 9.5 ± 6.8, and the mean number of mature oocytes per cycle was 7.4 ± 5.5. The obtained ongoing pregnancy per started cycle was 26.0% (95% confidence interval [CI]: 24.1-27.9) and the obtained ongoing pregnancy per puncture was 27.0% (95% CI: 25.0-29.0).CONCLUSIONS:This is the first cohort to describe Fertistartkit treatment management in real-life conditions. The real-world data show that Fertistartkit is an effective option for ovarian stimulation

Average clinical semen parameters in 239 male patients divided by %M3 SPTRX3 and pregnancy.

<p>The only group that shows somewhat reduced average clinical semen parameters are the men with highest SPTRX3 levels (>15% M3). Idiopathic infertility patients with apparently high SPTRX3 level (>15% idiopathic; bottom row) shows acceptable, normal clinical semen parameters. David's classification scheme for sperm morphology was employed <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061000#pone.0061000-Auger1" target="_blank">[32]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061000#pone.0061000-David1" target="_blank">[33]</a>. SE = standard error.</p

Diagrams of average SPTRX3 content in spermatozoa of men from 239 infertile couples.

<p>(<b>A</b>) Average pregnancy rates after infertility treatment (IVF or ICSI) decreased gradually with the increased sperm SPTRX3-content in 239 couples divided by % of SPTRX3 spermatozoa (flow cytometric %M3 value; x-axis) into subgroups with less than 5%, 5–9.99%, 10–14.99%, and more than 15% of SPTRX3-positive spermatozoa. Compared to men with less than 5% SPTRX3-positive spermatozoa, those carrying more than 15% of such cells had their chance of conceiving reduced by a half (41.2% pregnant, vs. 19.2% pregnant; one-sided p<0.05). (<b>B</b>) Low levels of sperm SPTRX3 were found in female and idiopathic infertility-couples, compared to male I an combined male & female infertility-couples. Different superscripts mark statistically significant differences between columns at p<0.001. The most significant difference was observed between female and male infertility couples (p<0.001). (<b>C</b>) Couples treated by IVF had significantly lower average SPTRX3-levels than those treated by ICSI, regardless of whether or not they achieved a pregnancy. Superscripts <i>^{a, b}</i> mark statistical difference at p<0.05. The most significant difference was found between pregnant IVF couples and pregnant ICSI couples (p<0.001). (<b>D</b>) Couples with female and idiopathic infertility had significantly lower SPTRX3 levels (average %M3-values) than couples previously diagnosed with male or combined (male and female) infertility. (<b>E</b>) Distribution of SPTRX3 levels within cutoffs in patients divided based on pregnancy outcome. Note that the couples delivering twins had the highest percentage of men with %M3 SPTRX below 15% and loweste percentage of men with >15% M3 SPTRX3.</p

SPTRX3 levels and ART outcomes in 72 male infertility patients divided by etiology.

<p>Most frequent etiologies included asthenozoospermia (A), astheno-teratozoospermia (AT), oligo-asthenozoospermia (OA), and oligo-astheno-teratozoospermia (OAT). One patient had obstructive azoospermia and one had obstructive oligo-teratozoospermia. Other, less frequent etiologies included teratozoospermia (2 patients), obstructive (1), vasectomy (1), varicocele (1), anti-sperm antibodies (2) and asthenozoospermia with anti-sperm antibodies (1). One couple reported failed IVF before being treated by ICSI. Couples diagnosed with combined male & female, female-only or idiopathic infertility excluded from this analysis/table.</p><p>A = asthenozoospermia.</p><p>AT = astheno-teratozoospermia.</p><p>OA = oligo-asthenozoospermia.</p><p>OAT = oligo-astheno-teratozoospermia.</p>*<p>Obstructive.</p>**<p>Other  =  teratozoospermia (2 patients), oligo-teratozoospermia, obstructive (1), vasectomy (1), varicocele (1), anti-sperm antibodies (2), asthenozoospermia with anti-sperm antibodies (1).</p>***<p>One couple reported failed IVF before being treated by ICSI.</p

Relationship between the clinically diagnosed male infertility and individual, conventional and flow cytometric SPTRX3 parameters reflective of sperm quality.

<p>An odds ratio smaller than 1 indicates that the parameter is negatively associated with the odds of being diagnosed with male infertility. A larger than 1 odds ratio indicates a positive relationship between the parameter and the clinical diagnosis of male infertility. An odds ratio larger than 1, e.g. 1.12 is interpreted as “the odds of being diagnosed with male infertility increase by 12% for every unit of increase in the parameter measurements.” An odds ratio smaller than 1, e.g., 0.014, is interpreted as “the odds of being diagnosed with male infertility decrease by 98.6% for every unit of increase in the measured parameter.”<u> Abbreviations: </u>C = sperm count; V = semen volume; M = total motility; PR = progressive motility; NX = morphology/% normal spermatozoa; NEC = percent of necrotic spermatozoa assessed by Williams test.</p