Using Small Non-Coding RNAs in Extracellular Vesicles of Semen as Biomarkers of Male Reproductive System Health: Opportunities and Challenges

Reproductive dysfunction and urogenital malignancies represent a serious health concern in men. This is in part as a result of the absence of reliable non-invasive tests of diagnosis/prognosis. Optimizing diagnosis and predicting the patient’s prognosis will affect the choice of the most appropriate treatment and therefore increase the chances of success and the result of therapy, that is, it will lead to a more personalized treatment of the patient. This review aims firstly to critically summarize the current knowledge of the reproductive roles played by extracellular vesicle small RNA components, which are typically altered in diseases affecting the male reproductive tract. Secondly, it aims to describe the use of semen extracellular vesicles as a non-invasive source of sncRNA-based biomarkers for urogenital diseases

Semen miRNAs Contained in Exosomes as Non-Invasive Biomarkers for Prostate Cancer Diagnosis

Although it is specific for prostatic tissue, serum prostate-specific antigen (PSA) screening has resulted in an over-diagnosis of prostate cancer (PCa) and many unnecessary biopsies of benign disease due to a well-documented low cancer specificity, thus improvement is required. We profiled the expression level of miRNAs contained in semen exosomes from men with moderately increased PSA levels to assess their usefulness, either alone or in addition to PSA marker, as non-invasive biomarkers, for the early efficient diagnosis and prognosis of PCa. An altered miRNA expression pattern was found by a high throughput profiling analysis in PCa when compared with healthy individuals (HCt) exosomal semen samples. The presence of vasectomy was taken into account for the interpretation of results. Fourteen miRNAs were selected for miRNA validation as PCa biomarkers in a subsequent set of semen samples. In this explorative study, we describe miRNA-based models, which included miRNA expression values together with PSA levels, that increased the classification function of the PSA screening test with diagnostic and/or prognostic potential: [PSA miR-142-3p miR-142-5p miR-223-3p] model (AUC:0,821) to discriminate PCa from BPH (Sn:91,7% Sp:42,9% vs Sn:100%Sp:14,3%); and [PSA miR-342-3p +/- miR-374b-5p] model (AUC: 0,891) to discriminate between GS >= 7 tumours and men presenting PSA >= 4 ng/ml with no cancer or GS6 tumours (Sn:81,8% Sp:95% vs Sn:54,5% Sp:90%). The pathway analysis of predicted miRNA target genes supports a role for these miRNAs in PCa aetiology and/or progression. Our study shows semen exosome miRNA-based models as molecular biomarkers with the potential to improve PCa diagnosis/prognosis efficiency. As the next step, further prospective studies on larger cohorts of patients are required to validate the diagnostic and/or prognostic role of the miRNA panel before it could be adopted into clinical practice

Impact of Extracellular Vesicle Isolation Methods on Downstream miRNA Analysis in Semen: A Comparative Study

Seminal plasma (SP) contains a unique concentration of miRNA, mostly contained in small extracellular vesicles (sEVs) such as exosomes, some of which could be clinically useful for diagnosis and/or prognosis of urogenital diseases such as prostate cancer (PCa). We optimized several exosome-EV isolation technologies for their use in semen, evaluating EV purifying effectiveness and impact on the downstream analysis of miRNAs against results from the standard ultracentrifugation (UC) method to implement the use of SP sEV_miRNAs as noninvasive biomarkers for PCa. Our results evidenced that commercial kits designed to isolate exosomes/EVs from blood or urine are mostly applicable to SP, but showed quantitative and qualitative variability between them. ExoGAG 3500× g and the miRCURY Cell/Urine/CSF 1500× g methods resulted as equivalent alternative procedures to UC for isolating exosomes/sEVs from semen for nanoparticle characteristics and quality of RNA contained in vesicles. Additionally, the expression profile of the altered semen sEV-miRNAs in PCa varies depending on the EV isolation method applied. This is possibly due to different extraction techniques yielding different proportions of sEV subtypes. This is evidence that the exosome-EV isolation method has a significant impact on the analysis of the miRNAs contained within, with important consequences for their use as clinical biomarkers. Therefore, miRNA analysis results for EVs cannot be directly extrapolated between different EV isolation methods until clear markers for delineation between microvesicles and exosomes are established. However, EV extraction methodology affects combined models (semen exosome miRNA signatures plus blood Prostate specific antigen (PSA) concentration for PCa diagnosis) less; specifically our previously described (miR-142-3p + miR-142-5p + miR-223-3p + PSA) model functions as molecular marker from EVs from any of the three isolation methods, potentially improving the efficiency of PSA PCa diagnosis

Evaluation of renographic and metabolic parameters in human Kidney transplantation

Background: the aim of this work is to demonstrate that the value of the mean transit time (MTT) obtained from the 99mTc-MAG3 renogram deconvolution is related to the levels of adenine nucleotides determined in cortical biopsies from transplanted kidneys. Methods: the functional state was estimated by means of the MTT and the initial height (H0) of the renal retention function obtained from the 99mTc-MAG3 renogram deconvolution and by the measure of adenine nucleotides obtained from biopsies. We studied 30 kidney graft recipients, 25 normal functioning grafts (NFG) and 5 with acute tubular necrosis (ATN). Results: the MTT is significantly longer for ATN (p < 0.001). The initial uptake values (H0) are significantly lower for ATN (p < 0.001). The sum of adenine nucleotides (SAN) is significantly greater for NFG than for ATN (p < 0.001). The values of the MTT seem to reflect the energy state of the cells in transplanted kidney. Conclusion: the analysis of MTT may be indicative of the functional metabolic recovery and thus it may be predictive of the renal graft function at least in the same extent than the biochemical analysis of a cortical renal biopsy immediately after blood reperfusion of the tissue

Percutaneous nephrolithotomy in the transplant kidney - a prospective 20 year multicentre experience

Introduction & Objectives: Percutaneous nephrolithotomy (PCNL) in transplanted kidneys presents unique endourological challenges. By pooling the collective experience of three centres, each combining high volume transplant and high volume endourology practices, we aim to accurately and reliably demonstrate the safety and feasibility of this technique

Endovascular management of hemorrhagic complications after percutaneous nephrolithotomy: 10-years experience

Background: bleeding is one of the most common and most important complications of percutaneous nephrolithotomy (PCNL), which is mainly controlled with conservative treatment options. Transcatheter arterial embolization is required in less than 1 % of the patients undergoing PCNL. There are only a few studies about endovascular treatment of vascular complications of PCNL. The purpose of this study was to evaluate renal arterial complications of PCNL and treatment outcomes with endovascular coil embolization. Patients and methods: this retrospective study evaluated 16 patients who underwent endovascular management for complications after PCNL, including diagnostic angiography. We analyzed the angiographic appearances of the vascular lesions that caused hemorrhages, treatment outcomes for endovascular coil embolization, and renal parenchymal loss rate following this treatment. Results: seven patients had a pseudoaneurysm, two patients had an arteriocaliceal fistula (ACF), five patients had a pseudoaneurysm and an arteriovenous fistula (AVF), and two patients had a pseudoaneurysm and an ACF. Of the 14 patients with pseudoaneurysms, five had more than one pseudoaneurysm. Endovascular coil embolization was successful in all patients, and it was able to stop the bleeding. After embolization, 12 patients had less than 10 % parenchymal loss, and 4 patients had 10-20 % parenchymal loss. Mean hospital stay after embolization was 2.3 ± 0.7 days (range, 1 to 3 days). Conclusions: the injuries seen in the intrarenal arterial system during the PCNL procedure can result in pseudoaneurysms and/or AVFs and/or ACFs, and more than one artery can be harmed. Arterial complications of PCNL can be treated with endovascular coil embolization while preserving renal function at a maximum level

Improving kidney retrieval from cDCD using normothermic extracorporeal membrane oxygenation

Introduction & Objectives: during the past decade, kidneys obtained after controlled circulatory death (cDCD) have significantlyincreased, in order to increase the number of donors. It achieved 24% of total Kidneys transplant during the last year in Spain. However, it implies warm ischemia times, resulting ina greater risk for organs. Focusing on graft quality optimization, abdominal Normothermic Extracorporeal Membrane Oxygenation (NECMO) has been implemented in order to restore blood flow before organ recovery. NECMO perfusion provides promising results for the use of liver grafts for cDCD. The aim of this study is to evaluate cDCD kidneys, obtained from NECMO technique compared with cDCD ultra-rapid retrieval

Hypothermic machine perfusion preservation after controlled donor cardiac death reduce delayed graft function

Introduction & Objectives: in Spain kidneys after controlled cardiac death (cDCD) has increased over the years. This group present a high incidence of DGF. Graft quality optimization has been the major interest over the last decade. The aim of this study is to evaluate de benefit of hypothermic machine perfusion (HMP) in reducing de delayed graft function (DGF) and primary non-function (PNF) compared to cold storage in cDCD. Other parameters are evaluated: 99mTc-MAG-3 scintigraphy TFS (tubular function slope) and ultrasound (US) resistive index (RI) 24-48 hours after surgery, rejection rate, duration of DGF. Materials & Methods: We are conducting a randomized prospective study since April 2017. We select all cDCD from one single institution, Hospital Universitari de Bellvitge, and we randomize 1:1 to cold storage and HMP. Data from donors, surgery, HMP, and post-operatory are collected; and analysed

Comparison of clinical outcomes between Maastricht-III kidney donors >65 years old and donors after brain death: a single center, matched-pair study

Introduction & Objectives: the inclusion of Maastricht Category-III (MIII) donors after circulatory death (DCD) has increased the donors' pool and, therefore, kidney transplants (KT). However, DCD have higher incidence of delayed graft function (DGF) compared to donors after brain death (DBD), being age one of its risk factors. Available data of using expanded criteria DCD is still controversial and conflicting in the current medical practice. The purpose of this study was to compare aged DCD outcomes to DBD regarding DGF, graft and patient's survival. Materials & Methods: We performed a retrospective observational matched-pair analysis of DCD >65 years old (yo) compared to DBD with minimum 1 year follow-up. Patients were matched according to donors' age (±2 years), receptors' age (±5 years), cold ischemia time (CIT) (±3 hours) and type of storage (cold vs perfusion machine). Stata14 program was used for statistical analysis: Tstudent, χ2 for descriptive analysis, logistic regression for DGF and Kaplan Meier survival curves

Multiple partial nephrectomy for multifocal synchronous renal cancer in a solitary kidney

Introduction & Objectives: multifocal synchronous renal cancer on a solitary kidney represent a challenging clinical scenario. The complexity of imperative nephron-sparring surgery in this setting resides in ensuring complete excision of cancer with the maximal preservation of renal function. We aim to present a case of multiple partial nephrectomy (MPN) for multifocal synchronous renal cancer in a patient with a solitary kidney and discuss our experience of imperative partial in this setting. Materials & Methods: We present a case of a 76 years old man with a past medical history of hypertension, chronic obstructive pulmonary disease, peripheral vascular disease, left radical nephrectomy for renal mass (2006) and a right renal artery stent placement for renal artery stenosis. During his surveillance, computerized axial tomography (CAT) scan showed 3 enhancing renal masses (2.2cm, 1.5cm and 1cm, respectively). Biopsy of the largest mass was consistent in clear cell renal cell carcinoma (ccRCC). Preoperative level of creatinine was 1.4mg/dL and estimated glomerular filtration rate (eGFR) 50ml/min/1,73m2. After ablative therapy was deemed unsafe, a MPN was planned