Research on the potential use of seminal fluid as a liquid biopsy for prostate cancer detection has been limited due to challenges associated with acquisition of this bodily fluid in clinical studies. Here we sought to expand on our previous analysis, which demonstrated high levels of prostate-derived cell free DNA (cfDNA) in seminal fluid in presumed healthy individuals, to a much larger cohort that included participants with prostate cancer. A total of 279 men scheduled for prostate biopsy were enrolled over 4 months across 12 sites. Prior to their biopsy, participants mailed a seminal fluid sample collected at home to the laboratory, from which cfDNA was extracted and underwent methylation analysis. Consistent with our earlier study in healthy individuals, we observed an abundance of high molecular weight (HMW) cfDNA in all samples. Tissue-of-origin deconvolution revealed that granulocytes and sperm were the principal contributors to seminal fluid cfDNA, while prostate-derived cfDNA was present at abundances readily detectable with current technologies. The nucleosomal fraction was very pronounced in some but not all samples and was determined to be correlated with the relative sperm signal. The sperm signal was also observed to be associated with an increase in small insert sizes (< 125 bp) in the sequenced libraries. Unsupervised clustering revealed two distinct populations driven by the abundance of sperm and granulocytes. Since summarizing at the genomic region level confounded tissues of different origins, fragment-level DNA methylation features were used to characterize and quantify the prostate cancer related signal, and features associated with clinically significant prostate cancer were identified. This study expands on our previous work to further characterize seminal fluid and highlights its potential as a promising liquid biopsy medium for the detection and monitoring of clinically significant prostate cancer.

e17135 Background: While Prostate Specific Antigen (PSA) is the standard of care for prostate cancer screening, it is highly controversial due to the performance characteristics of the test. Magnetic Resonance Imaging (MRI) is becoming increasingly used as a reflex test for high PSA, but there are limitations with cost, disparities in access, variations in interpretation, and moderate sensitivities/specificities. Given that the prostate is a reproductive organ and a large portion of seminal emissions are generated from the prostate, this study aimed to investigate whether a novel prostate cancer screening platform using cell free nucleic acids from seminal plasma could improve the detection of clinically significant prostate cancer. Methods: Over a 4-month period, 276 patients scheduled to undergo a prostate biopsy were enrolled across 12 sites. Semen samples were collected at home by men aged 40 yrs and older and shipped to the Fellow Health laboratory. Seminal plasma was prepared by centrifugation and cell free nucleic acids extracted. DNA methylation libraries were prepared using the Twist methylome target enrichment panel and RNA libraries were prepared using Watchmaker kits. All libraries were sequenced on a NovaSeq X. Clinical data (including PI-RAD score, cancer status and Grade Group (GG)) were merged with the sequencing results and machine learning tools employed for modeling predictive algorithms. Results: Median age was 64 (IQR=10 years), median PSA was 6.1 (IQR=3.925ng/ml), 79% of subjects were white, 45% had BPH, 12.5% were vasectomy positive and 66% received MRI prior to biopsy. 180 valid semen samples were received prior to the cutoff date, of which 146 were evaluable for both RNA and DNA analysis. The well-known prostate tissue markers PSA and TMPRSS2 were highly abundant in seminal fluid with mean normalized transcripts per million of 1597 and 397.5 respectively. Using healthy controls to set the limit of blank, 45.1% of cancer patients harbored an ETS fusion (ERG, ETV1, ETV4, ETV5, FLI1). The data was split into a training set (n=115) and an independent test set (n=31). With a panel of 66 expression markers, 26 methylation markers and PSA, the AUC in the test set was 0.83 for differentiating clinically significant prostate cancer (GG2+) from no cancer (<=GG1). Conclusions: In this cohort, the use of MRI still resulted in unnecessary biopsies 69.4% of the time. Meanwhile, we demonstrate that our multiomic approach using cell free DNA and RNA achieves high accuracy for identifying clinically significant prostate cancer. At-home semen collection offers a convenient alternative to in-lab testing and may have the potential to augment MRI in post-PSA reflex testing.

PURPOSE:The AUA recommends postvasectomy semen analysis (PVSA) to verify successful vasectomy. However there remains poor patient compliance. We sought to assess whether mail-in PVSA improves patient compliance across a wide range of practice types. MATERIALS AND METHODS:Prospective data were collected on all men who received a Fellow PVSA kit between April 2021 and August 2023 in a nationwide cohort. Date of kit activation, practice type, clinic zip code, and date of kit accession/processing at the lab were collected. Compliance rates for each practice area were reported. χ2 tests of independence, logistic regression models, and multivariable logistic analysis were performed to assess the impact of relevant variables. RESULTS:Overall compliance across all practice areas was 69% following an 18-week period of observation (n = 16,105) and 82% (n = 6687) following a 40-week period. Compliance rates were highest and similar for small urology practices (<5 providers), including Veterans Affairs practices, ranging from 76% to 82% at 18 weeks to 85% to 87% at 40 weeks. Large urology practices had slightly lower compliance rates with 66% at 18 weeks to 80% at 40 weeks. The univariable logistic regression model demonstrated that patients in small urology practices have a 63% greater odds of 26-week compliance, on average, compared to those who receive care in large urology practices (odds ratio 1.63, 95% CI 1.48-1.79). CONCLUSIONS:Fellow's mail-in PVSA offers improved PVSA compliance over previously published data. Improved compliance is seen across all practice types. Despite these successes, there is significant room for improvement to achieve 100% compliance.

Abstract Introduction Undergoing fertility testing can be a stressful event in a couple's life. This can impact multiple aspects of life, including sex. Objective To evaluate the relationship between stress and sexual function in men undergoing fertility testing. Methods We performed a review of a prospectively maintained database of men utilizing a mail-in semen analysis test system developed by Fellow Health, Inc. Those included consented to participate in research, were 18 or older, and completed survey questionnaires on both their quality of life and sexual function. Stress level (low, moderate, or high stress) was determined using the validated Perceived Stress Scale (PSS) 10-question survey. The sexual function survey assessed libido, orgasmic and ejaculatory dysfunction, intercourse frequency, Erection Hardness Score, and IIEF-5 score. Descriptive statistics were reported and chi-squared and t-tests were used to determine statistical significance at the 0.05 alpha level. Results A total of 2077 men undergoing fertility testing were included in this study. 50.2% reported moderate stress levels and 2.6% reported high stress levels. Those with low stress were older than those with moderate or high stress (average 35.4 vs. 34.3 and 34.0 years, respectively; p < 0.01). Men with high stress levels were more likely to report a “poor” or “terrible” libido (31.4%) compared to those with moderate or low stress, 17.2% and 9.5%, respectively (p < 0.01). Stress was highly associated with weekly intercourse frequency (p < 0.01): among men with high levels of stress, 25.5% reported having sex less than once a week compared to 6.0% of men with low stress. Likewise, no highly stressed men reported having sex more than six times per week compared to 2.8% of men with low stress. Whereas 80.4% of men with low stress were evaluated to have “No signs of ED” by the IIEF-5, the same was true for only 62.3% of moderately stressed men and 56.0% of highly stressed men (p < 0.01). Conclusions More than half of men undergoing fertility testing report increased levels of stress, which is negatively associated with libido and erectile function. In men seeking fertility evaluation, lifestyle factors such as stress levels should be assessed to identify patients who may benefit from stress management strategies. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Fellow Health.

e13524 Background: There are many circumstances under which someone may choose to bank their sperm, but forthcoming cancer treatment is among the most common. The advent of services offering mail-in semen analysis and cryopreservation has expanded access to sperm banking, especially for those living in rural areas, far away from traditional andrology labs. This study summarized demographic characteristics and reasons for cryopreservation among those using one such US-based mail-in service, with a focus on patients banking sperm prior to cancer treatment. Methods: Patients >18 years who banked sperm using a mail-in cryopreservation solution developed by Fellow Health, Inc. from March 2021 to January 2024 were included. Test kits sold for research purposes were excluded. To categorize each patient by reason for cryopreservation, we made use of a patient-provided reason, supplied during test kit registration, where available. For the remaining patients, reason was inferred using several data sources including medical history, medications, and clinic and physician information. Patients who could not be reliably classified were omitted from this portion of the analysis. Descriptive statistics of key demographics and reason for cryopreservation were reported under an IRB-approved protocol. Results: 2,154 patients met our inclusion criteria and 725 (33.7%) provided demographic information. Average age was 34.7 (SD=9.2) years and 11.9% identified as a trans female. The majority of the population identified as white (79.5%) and non-Hispanic (84.8%). Over half of the population had a college degree or greater (59.3%), was married/partnered (57.1%), and had private insurance (84.9%). Among the 756 patients whose reason for cryopreservation was discernible, the majority, 450 (59.5%), were doing so in the context of fertility treatment, with only 46 (6.1%) indicating cancer treatment. Other reasons included pre-vasectomy storage (n=137, 18.4%) and gender affirming hormone therapy (n=90, 11.9%). Of the patients who indicated cancer treatment as their reason for cryopreservation, 50.0% lived greater than a 25-mile drive of the nearest National Cancer Institute (NCI)-designated Cancer Center. Conclusions: Surprisingly, few patients utilizing a mail-in service for cryopreservation did so prior to beginning cancer treatment, with most originating from the fertility or pre-vasectomy context. Use was disproportionately high among trans women initiating hormone replacement therapy (HRT), perhaps demonstrating their preference for at-home sample collection to avoid the unique challenges they encounter in a clinical setting. Especially for oncology patients who are admitted to a hospital and cannot leave for fertility preservation, there is an opportunity to further integrate mail-in cryopreservation services within oncology to improve access and reduce time costs.

Abstract Introduction Depression is a common mental health disorder affecting men of reproductive age. Depression or depressive symptoms may impact a variety of health factors, including sexual function. Trying to conceive may contribute to additional stress and negative emotions for the couple. Objective To determine the rate of sexual dysfunction in men with depression who are undergoing fertility testing. Methods A review was performed of a prospectively maintained database of men utilizing a mail-in semen analysis test system developed by Fellow Health, Inc. Men ≥18 years were included if they consented to participate in research and completed survey questionnaires on both their quality of life and sexual function. Depression level (normal, mild, moderate, or severe) was derived via the short-form depression assessment developed by the Patient Reported Outcomes Measurement Information System (PROMIS). The sexual function survey assessed libido, orgasmic and ejaculatory dysfunction, intercourse frequency, Erection Hardness Score, and IIEF-5 score. Descriptive statistics were reported and chi-squared tests were used to determine statistical significance at the 0.05 alpha level. Results Of the 2125 men included, 15.2% reported mild depression and 12.8% moderate or severe depression. The presence of depression was negatively associated with libido, with 11.2% of men with normal levels of depression reporting terrible or poor libido compared to 18.7% with mild depression and 22.2% with moderate or severe depression, p < 0.001. Compared to men without depression, those with moderate or severe depression were nearly twice as likely to report ejaculatory dysfunction (17.2% vs. 29.7%) and three times as likely to report orgasmic dysfunction (3.9% vs. 13.0%), p < 0.001 for all. Severity of depression was also predictive of an erection sufficient for penetration: whereas 96.8% of those with normal levels of depression reported an erection sufficient for penetration, the same could be said for only 94.5% of those with mild depression and 92.5% of those with moderate or severe depression (p < 0.001). Only 53.1% of men with moderate or severe depression had an IIEF-5 score greater than 21 (i.e., “No signs of ED”), compared to 60.9% with mild depression and 75.8% without depression, p < 0.001. Importantly, men with moderate to severe depression were more than twice as likely to report intercourse frequency less than once weekly (18.0%) as opposed to men without depression (7.4%), p < 0.001. Conclusions The presence of depression in men undergoing fertility testing is negatively associated with libido, ejaculatory, orgasmic, and erectile function. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Fellow Health.