Parallel Meeting on Wednesday 3, September 2025, 17:30 - 18:30
17:30 - 17:34
S07-1 (OP)
Kholoud AL-SHAFAI 1, Seem ARAR 2, Asma JAMIL 2, Amina AZZAH 2, Luis SARAIVA 1 and Tariq Osman ABBAS 3
1) Sidra Medicine, Translational Medicine, Doha, QATAR - 2) Sidra Medicine, Division of Paediatric Surgery,, Doha, QATAR - 3) Sidra Medicine, Urology, Doha, QATAR
PURPOSE
To identify genetic variants associated with hypospadias in familial cases, a congenital condition with significant genetic underpinnings but largely unexplored specific causes. Understanding these genetic contributors could improve diagnosis, genetic counseling, and targeted interventions.
MATERIAL AND METHODS
We conducted a familial study using a biobank, collecting biospecimens from 27 subjects across seven families with a history of hypospadias. Whole genome sequencing (WGS) was performed to identify potential pathogenic variants. GEMINI bioinformatics tools were used to analyze and assess variant segregation within families.
RESULTS
Analysis revealed several pathogenic variants linked to hypospadias. A TTC37 variant was associated with coronal hypospadias in both the patient and his father. An OBSL1 variant was identified in the patient, father, and brother with scrotal hypospadias. A EIF2B5 variant was found in the father, despite no signs of Leukoencephalopathy. Additionally, an autosomal dominant INO80 variant appeared in multiple family members, and a homozygous COL6A3 variant was observed in the patient's brother with increased protein expression. Furthermore, splice site variants in LHFP and ACADVL were identified in affected family members, indicating a complex genetic landscape for hypospadias inheritance.
CONCLUSIONS
This study uncovers multiple genetic variants associated with hypospadias, highlighting the condition's genetic complexity. These findings could aid genetic counseling and the development of targeted interventions. Future studies should further investigate the functional role of these variants in hypospadias.
17:34 - 17:38
S07-2 (OP)
Yu DING 1, Zhiwei PENG 2, Yiqing LYU 2 and Fang CHEN 2
1) Shangai Children's Hospital, Department of Urology, Shanghai, CHINA - 2) Children's Hospital of Shanghai, Urology, Shanghai, CHINA
PURPOSE
The formation of hypospadias is affected by prenatal nutrition and hormone levels. The placenta is the sole source of nutrients for the fetus and secretes a variety of essential hormones and cytokines. However, the precise role of placental function in this process remain unknown.
MATERIAL AND METHODS
Integrated transcriptomic and metabolomic profiling of placental tissues from controls and hypospadias patients identified Zinc Finger Protein 511 (ZNF511) as a critical transcriptional regulator of ribosome-related gene clusters, disrupting transporter synthesis and placental function. Single-cell RNA sequencing delineated pivotal cellular subpopulations. Furthermore, proteomics, IP, ChIP-seq, and other techniques elucidated the mechanistic role of ZNF511 in ribosome biogenesis. A placenta-specific ZNF511 knockout murine model subsequently confirmed male fetal phenotypic anomalies.
RESULTS
We found that ZNF511 functions as a transcription factor that binds to ribosomal gene (RPS and RPL) promoters, recruits small nucleolar ribonucleoprotein (snoRNP) complexes to nucleolar regions, and enhances RNA polymerase I activity to coordinate rRNA transcription/processing, thereby augmenting ribosome biogenesis efficiency and affecting the synthesis of downstream transport proteins. This process predominantly occurs in placental syncytiotrophoblasts, where ZNF511 downregulation in hypospadias placentas disrupts ribosomal homeostasis, culminating in metabolic dysregulation. Furthermore, ZNF511 expression is transcriptionally regulated by the mTOR signaling pathway. Notably, placenta-specific ZNF511 knockout mice exhibited preterm delivery, low birth weight, and male offspring hypospadias phenotype.
CONCLUSIONS
ZNF511 in placental trophoblast cells plays a crucial role in ribosome biosynthesis and fetal nutrient hormone levels, providing significant insights into the underlying pathogenesis of hypospadias as well as genetic counseling.
17:38 - 17:42
S07-3 (OP)
Yiqing LYU, Fang CHEN, Yichen HUANG and Yu DING
Shanghai Children's Hospital, Urology, Shanghai, CHINA
PURPOSE
Exosomes secreted from urine-derived stem cells (USC-exosomes) play a significant role in promoting vascular regeneration. We investigates the use of USC-exosomes to induce the regeneration of spongiosum beneath the neourethral in a rabbit model.
MATERIAL AND METHODS
A rabbit model with a 1.5×1.0 cm urethral defect was created and repaired using buccal mucosa. Three rabbits were assigned to experimental group and received USC-exosomes at a concentration of 1-3×10^10/ml, with a dosage of 0.5 ml/cm^2, applied to the outer surface of the neo-urethra. Three served as control group, receiving sodium hyaluronate in the same manner. Urethrography was performed three months later. Histopathological studies, including HE staining, Masson's trichrome staining, and immunostaining for α-SMA, were conducted to evaluate the restoration of the spongiosum. While the urethral tissues were sequenced and screened by qRT-PCR to find the differentially expressed genes(DEGs), then analyze DEGs by Gene Ontology and KEGG database to find the internal mechanism.
RESULTS
Urethrography revealed the diameter of the urethra in experimental group was significantly wider. In experimental group, the urethral outer wall displayed prominent sinusoidal vascular structures and smooth muscle fibers, whereas the control group's urethral wall consisted mainly of fibrous connective tissue. Histopathological studies showed that experimental group displayed significantly greater sinusoid formation, a higher density of smooth muscle fibers, and enhanced α-SMA expression compared to control group. Bioinformatics analysis found the DEGs mostly related to angiogenesis between the two groups focused on the ECM receptor interaction pathway and PI3K/Akt signaling pathway, especially on down-regulate of PTEN and up-regulate of Akt3, CREB5, BCL2, ITGA4 and COLIA2.
CONCLUSIONS
USC-exosomes induced the formation of spongiosum-like structures on the outer wall of the urethra. This study underscores the potential to reconstruct a more physiologically normal urethra with regenerated corpus spongiosum.
17:42 - 17:46
S07-4 (OP)
Yichen HUANG, Fang CHEN, Yiqing LYU, Ming WU, Yu DING and Zhiwei PENG
Shanghai Children's Hospital, Urology, Shanghai, CHINA
PURPOSE
We previously found that the pathological changes of the corpus spongiosum (CS) in hypospadias mainly concentrated in smooth muscle tissue, presenting as phenotype transformation from contractile to synthetic, which might be related to frequent postoperative complications. The role of low-intensity pulsed ultrasound (LIPUS) in regulating vascular smooth muscle cells and angiogenesis has been confirmed.
This study is aimed to demonstrate the feasibility of regulating phenotype transformation of CS smooth muscle cells (SMCs) in hypospadias by LIPUS and explore the potential mechanisms.
MATERIAL AND METHODS
The SMCs were extracted from CS tissue in patients with proximal hypospadias. Appropriate LIPUS irradiation intensity and duration which could regulate phenotype transformation of CS SMCs were determined in vitro. Then 71 patients with severe hypospadias were randomly divided into control group and LIPUS group to verify the phenotype regulation of LIPUS on CS smooth muscle in vivo. At last, the potential mechanisms were explored in vitro.
RESULTS
In vitro experiments showed that LIPUS with an intensity of 0.1w/cm2 and duration for 10 minutes could significantly increase the expression of contraction markers in CS SMCs and decrease the expression of synthesis markers. Moreover, LIPUS stimulation could alter the phenotype of SMCs in patients with proximal hypospadias. RNA sequencing results revealed that PPAR-r was significantly increased after LIPUS stimulation. After over-expression of PPAR-r, the contraction markers of CS smooth muscle cells significantly increased, and knockdown of PPAR-r blocked this effect.
CONCLUSIONS
LIPUS could regulate CS SMCs from synthetic type to contractile in hypospadias, showing promising in postoperative tissue remodeling. The PPAR-r signaling pathway might play an important role in this process.
17:58 - 18:02
S07-5 (OP)
Nao IGUCHI 1, Anna MALYKHINA 1 and Duncan WILCOX 2
1) University of Colorado School of Medicine, Surgery, Aurora, USA - 2) Children's Hospital Colorado, Pediatric Urology, Aurora, USA
PURPOSE
This study aimed to investigate the pathological mechanisms behind bladder and bowel dysfunction (BBD), a common yet often underdiagnosed pediatric condition marked by lower urinary tract symptoms (LUTS) and abnormal bowel habits like constipation and/or encopresis. BBD is known to cause long-term complications such as recurrent UTIs, vesicoureteral reflux, and renal scarring, but the mechanisms behind these links remain unclear. We hypothesize that bowel dysfunction induces bladder-bowel cross-organ sensitization via overlapping neural pathways and tissue changes, including inflammation, fibrosis, and alterations in cellular signaling and connectivity.
MATERIAL AND METHODS
With IACUC approval, functional constipation was induced in juvenile male mice by surgically narrowing the external anal sphincter. Sham-operated mice served as controls. Four days post-surgery, the effects of constipation were evaluated using in vivo urodynamic studies and in vitro physiological assessments of bladder strips. Additionally, quantitative RT-PCR was used to analyze gene expression patterns in the bladder and lumbosacral dorsal root ganglia (LsDRG).
RESULTS
Constipation caused detrusor overactivity, shown by increased urinary frequency and spontaneous contractions. Gene expression analysis revealed a 1.5-fold upregulation of excitatory serotonin receptors (Htr2a, Htr2c) in the bladder (p<0.05). Serotonin enhanced spontaneous activity in bladder strips from constipated mice, increasing frequency and force by 1.6- and 2.4-fold, respectively (p<0.0001). These effects were reversed by ketanserin, an Htr2 antagonist. Additionally, the LsDRG in constipated mice showed a 1.6- to 1.8-fold increase in genes modulating sensory activation, including serotonin receptors (Htr1a, Htr3) and Mrgprb2 (p<0.05).
CONCLUSIONS
The findings highlight how constipation alters sensory pathways, contributing to detrusor overactivity and increased bladder activity. Upregulation of serotonin receptors and sensory modulators plays a key role in bladder dysfunction, suggesting these receptors as potential therapeutic targets for children with BBD.
18:02 - 18:06
S07-6 (OP)
Venkat RAMAKRISHNAN 1, Alexander BIGGER-ALLEN 1, Kyle COSTA 1, Gabriel-Luis OCAMPO 1, Hatim THAKER 1, Lina DAI 1, Andrea SARTORI 2, Maryrose SULLIVAN 3, Vivian CRISTOFARO 3, Margot DAMASER 4, Siam OOTTAMASATHIEN 1, Carlos ESTRADA 1, Michael CIMA 5 and Rosalyn ADAM 1
1) Boston Children's Hospital and Harvard Medical School, Urology, Boston, USA - 2) Beth Israel Deaconess Medical Center and Harvard Medical School, Medicine, Boston, USA - 3) Boston Veterans Affairs Medical Center and Harvard Medical School, Surgery, West Roxbury, USA - 4) The Cleveland Clinic and Cleveland Veterans Affairs Medical Center, Urology, Cleveland, USA - 5) Massachusetts Institute of Technology, Materials Science and Engineering, Cambridge, USA
INTRODUCTION
The management of neurogenic lower urinary tract dysfunction (NLUTD) requires low bladder pressures to protect the upper urinary tract. Oral anticholinergics such as trospium chloride (TrCl) are a mainstay of therapy but face challenges with adherence and systemic side effects. Intravesical TrCl delivery achieves high local concentrations with minimal systemic absorption and is efficacious in idiopathic detrusor overactivity, though its effects on NLUTD remain unexplored. We examined the effect of daily intravesical TrCl instillation on bladder function in a rodent spinal cord injury (SCI) model.
METHODS
Ten-week-old female Sprague-Dawley rats underwent suprapubic catheter implantation, followed by T8–T10 spinal cord transection seven days later. After randomization into control (water; n=3) or TrCl (0.65 µg/mL; n=3) treatment groups, daily 500 µL intravesical instillations began the following day. After four weeks, bladder function was evaluated using conscious cystometry (60 µL/min saline infusion) for up to 2 hours. Statistical analysis was performed using the Dunn test.
RESULTS
Of the overall cystometry time, TrCl bladders demonstrated pressures less than 20 cm H2O significantly more often (88.4% of the time vs. 39.2% in controls; p < 0.05), and greater than or equal to 20 cm H2O significantly less often (11.6% vs. 60.7%; p < 0.05). This trend continued when assessing truly pathologic pressures greater than 40 cm H2O (0.736% vs. 27.4%; p = 0.121). The AUC was significantly reduced by TrCl treatment (3.9x104 vs. 14.3x104; p < 0.05).
CONCLUSIONS
Daily intravesical TrCl instillation in SCI rats significantly reduced overall bladder pressures compared to untreated animals, pointing to a potential morbidity-reducing additional treatment avenue for an otherwise challenging patient population. Further validation of additional urodynamic parameters, systemic TrCl distribution, and mechanistic changes is required.
18:14 - 18:18
S07-7 (OP)
Marios MARCOU 1, Arndt HARTMANN 2, Helge TAUBERT 3 and Sven WACH 3
1) Barmherzige Brüder Klinik St. Hedwig, Klinik für Kinderurologie, Regensburg, GERMANY - 2) University Hospital Erlangen, Institute of Pathology, Erlangen, GERMANY - 3) University Hospital Erlangen, Clinic of Urology and Pediatric Urology, Erlangen, GERMANY
PURPOSE
Lichen sclerosus (LS) is a chronic, inflammatory skin condition that affects both children and adults, with the genital region being the primary site of manifestation. Despite its clinical relevance, molecular mechanisms underlying the disease in male pediatric patients remain poorly understood. This study aimed to identify and analyze the differential expression of microRNAs (miRNAs) in LS tissue compared to adjacent normal tissue and non-LS controls to uncover potential diagnostic and therapeutic biomarkers.
MATERIAL AND METHODS
Archival foreskin tissue samples were obtained from 98 pediatric circumcision cases, including 41 histologically confirmed LS cases. LS and adjacent normal tissues from LS patients, as well as normal tissue from non-LS patients, were analyzed. RNA was isolated from formalin-fixed paraffin-embedded (FFPE) tissue, and quantitative real-time PCR (qRT-PCR) was performed to evaluate the expression of nine miRNAs previously implicated in LS or related conditions.
RESULTS
Four miRNAs (hsa-miR-146a-5p, hsa-miR-146b-5p, hsa-miR-150-5p, and hsa-miR-155-5p) were significantly upregulated, and two (hsa-miR-199b-5p and hsa-miR-200b-3p) were significantly downregulated in LS tissues compared to adjacent normal tissues and non-LS controls (all p < 0.001). ROC analysis revealed hsa-miR-155-5p and hsa-miR-150-5p as the most robust diagnostic markers, with area under the curve (AUC) values of 0.892 and 0.864, respectively, when differentiating LS from normal tissue. Additionally, pediatric LS tissues exhibited fewer significant miRNA correlations compared to normal tissues, suggesting a disruption of regulatory networks. Age-specific analysis revealed higher expression of hsa-miR-150-5p and increased inflammation in prepubertal patients (≤9 years).
CONCLUSIONS
This study demonstrates distinct miRNA expression patterns in male pediatric LS patients, identifying hsa-miR-155-5p and hsa-miR-150-5p as promising diagnostic biomarkers. The findings highlight miRNAs as potential therapeutic targets and underline their role in the molecular pathogenesis of pediatric LS, warranting further investigation into their regulatory pathways and interactions with hormonal and inflammatory mechanisms.
18:18 - 18:22
S07-8 (OP)
David KOOS 1, Matthew THORNTON 2, Paola AGUIARI 1, Valentina VILLANI 1, Hripsime CHOMOYAN 1, Brendan GRUBBS 2, Roger DE FILIPPO 3, Stefano DA SACCO 3, Laura PERIN 3 and Astgik PETROSYAN 4
1) Children's Hospital Los Angeles, Los Angeles, USA - 2) University of Southern California, Los Angeles, USA - 3) Children's Hospital Los Angeles / Keck School of Medicine USC, Los Angeles, USA - 4) Children's Hospital Los Angeles / Keck School of Medicine USC, Urology, Los Angeles, USA
PURPOSE
The extracellular matrix (ECM), a vital tumor microenvironment (TME) component, significantly influences cancer progression and therapeutic resistance. Wilms tumor (WT), a pediatric renal cancer arising from abnormal kidney development, exhibits altered ECM architecture, which may drive tumor progression. This study aims to characterize ECM changes in WT compared to normal kidney tissue and investigate their impact on cancer cell behavior and tumorigenic potential.
MATERIAL AND METHODS
WT and normal kidney tissue samples were decellularized, and ECM architecture was visualized using second-harmonic generation (SHG) microscopy and immunofluorescence staining. Cancer BioMarker Antibody Array measured cancer-related protein expression in the ECM. Cancer and normal cells were seeded on decellularized WT and normal ECM scaffolds for 21 days, with migration and proliferation monitored using STELLARIS DIVE multiphoton microscopy. Bulk RNA sequencing and single-cell ATAC-seq analyzed transcriptomic changes and chromatin accessibility.
RESULTS
WT ECM showed fragmented, disorganized collagen and increased inflammatory markers (e.g., alpha-1 antichymotrypsin), cytoskeletal remodeling proteins (e.g., tubulin alpha), and EMT regulators (e.g., TGFB1). Cancer cells seeded on WT ECM upregulated EMT genes (FOXC2, TGFB2) and ECM remodeling genes (ADAMTS5), while downregulating cell cycle regulators (PPM1L) and integrin interaction genes (COL8A2). Normal cells on WT ECM displayed increased cancer-associated gene expression (e.g., AKT3, BCL2) and decreased integrin-related genes (COL6A3, ICAM1). Single-cell ATAC-seq revealed WT-specific cancer stem cell populations expressing EMT transcription factors and unique ECM signatures.
CONCLUSIONS
WT ECM alterations, including collagen disruption and EMT promotion, foster a tumor-supportive environment. Targeting ECM-driven mechanisms may offer novel therapeutic strategies to disrupt tumor progression.