Citations

Abstract Regulation of gene expression is central to the development of immune cells and their ability to respond to infection. As part of a clinical evaluation, we identified two sisters with recurrent infections, hypogammaglobulinemia, and memory B cell deficiency, diagnosed as common variable immunodeficiency. Whole exome sequencing identified a heterozygous variant (Leu50Ser, L50S) in a conserved region of EZH2, the catalytic subunit of the epigenetic gene repressor Polycomb Repressive Complex 2 (PRC2). EZH2-catalyzed histone H3 lysine 27 methylation (H3K27me) in bulk was not overall significantly disrupted by this variant, in patient samples or cell lines expressing EZH2-L50S. EZH2-L50S protein is expressed similar to wild-type and can form PRC2. However, we find that specific genomic regions that normally have high wild-type levels of H3K27me3 are deficient in the L50S context, particularly around gene promoters. EZH2-L50S is still recruited to these sites, but is not as active. Using recombinant purified PRC2, we determine that L50S affects methylation of nucleosomes and disrupts allosteric stimulation that normally amplifies H3K27me3, consistent with the location of L50 in the allosteric regulatory region of PRC2. Thus, variation of EZH2 L50, occurring at low frequency in the population may interfere with normal B cell gene expression patterns, contributing to immunodeficiency. This study has implications for genetic variation in PRC2 in the general population.

Abstract Multiple myeloma is a malignancy of clonally expanded plasma cells shaped by complex interactions with the immune microenvironment (IME). To investigate immune correlates of treatment response and disease progression, we conducted multi-omics profiling including CD138neg single-cell RNA sequencing of 243 bone marrow samples from 102 patients (631,226 cells) and CD138pos bulk RNA and whole-genome sequencing from 209 samples. In longitudinal analyses, interferon-γ signaling associated with markers of impaired T-cell memory after autologous stem cell transplant, whereas naïve B-cell abundance and immunoglobulin diversity correlated with improved progression-free survival (HR = 0.48; P = 2.3e−4). At disease progression, multiple myeloma cells upregulated cancer–testis antigens (CTAg) and immune effector genes, with concurrent B-cell depletion, enrichment of myeloid-derived suppressor cell expression, and phenotypic T-cell exhaustion. These findings highlight dynamic immune–tumor interactions, identifying naïve B-cell reconstitution as a biomarker of durable response and CTAgs as potential targets for high-risk disease at progression. Significance: Longitudinal profiling of multiple myeloma and the IME revealed dynamic immune–tumor interactions across the disease course. Altered expression in CD8+ T cells limited memory phenotype after transplant, whereas naïve B-cell recovery associated with sustained treatment response. At progression, CTAg expression associated with immunosuppression, revealing novel mechanisms of immune dysregulation.

BACKGROUND Among antiretroviral therapy–suppressed (ART-suppressed) people with HIV (PWH), women have higher levels of some inflammatory markers than men, but the effect of sex on the inflammatory proteome, and whether age modifies these differences, remain unclear. METHODS Plasma inflammatory protein levels were assessed in ART-suppressed PWH from the Center for AIDS Research Network of Integrated Clinical Systems. The relationship between sex and plasma proteins — including 22 interferon-α response pathway proteins — was assessed, adjusting for confounders and assessing interactions by age. RESULTS Of 922 participants, 162 (18%) were female. The median age was 47, above which the majority of women had undetectable plasma anti-Müllerian hormone levels, a biomarker of ovarian reserve. Age modified the influence of sex on the inflammatory proteome. Older age (>47) was associated with greater increases among women than men in 194 proteins. Interferon-α response proteins were higher in men in those ≤ 47 but higher in women in those > 47 (interaction P < 0.001). Among the 131 proteins associated with mortality risk ( q < 0.05), only 5 differed by sex among those ≤ 47, while 79 differed by sex in those > 47, with nearly all being higher in women. Women had decreased mortality than men ≤ 47 ( P < 0.001) but had similar mortality > 47 ( P = 0.84). CONCLUSION The menopausal transition appears to increase systemic type I interferon responses and inflammation in women with HIV, which may contribute to a loss of female advantage in mortality. FUNDING NIH National Heart, Lung, and Blood Institute; National Institute of Neurological Disorders and Stroke; National Institute of Allergy and Infectious Diseases.

Key Points The kidney microvasculature was quantified reliably in human autosomal dominant polycystic kidney disease early on, using super-resolution ultrasound localization microscopy. Microvascular parameters correlated with structural and functional indices, constituting promising biomarkers to evaluate noncystic parenchyma. Microvascular parameters may serve to assess disease severity and monitor progression from the early stages of the disease. Background In vitro studies in individuals with autosomal dominant polycystic kidney disease (ADPKD) have shown extensive intrarenal microvascular remodeling. However, analyses have been limited to the time of kidney failure because of the invasive nature of current methodologies to assess the intrarenal microvasculature. We hypothesized that super-resolution ultrasound microvascular imaging, known as ultrasound localization microscopy, enabled by high-framerate ultrasound imaging and microbubble localization and tracking postprocessing techniques, would safely, reliably, and reproducibly assess the intrarenal microvasculature in individuals with ADPKD at an early stage of the disease. Methods We prospectively enrolled young (<40 years of age), well-characterized, early-stage (eGFR >90 ml/min per 1.73 m 2 ) individuals with ADPKD, and sex–age 1:1 matched controls ( n =17 each) and assessed the intrarenal microvasculature using ultrasound localization microscopy. We determined the intersonographer reliability, intraobserver repeatability, and interobserver reproducibility of microvascular parameters in both groups, and the day-to-day variability in the control group. Microvascular parameters were correlated with structural (total kidney volume) and functional (eGFR) markers in individuals with ADPKD. Results Ultrasound localization microscopy provided a super-resolved map of the intrarenal microvasculature in ADPKD and matched controls. Intrarenal microvascular parameters ( e.g ., vessel density) were quantified in a safe, reliable (intraclass correlation coefficients, 0.96; 95% confidence interval, 0.91 to 0.98), reproducible (intraclass correlation coefficient, 0.96; 95% confidence interval, 0.93 to 0.98), and repeatable (coefficient of variations, 3.6%) manner. In control individuals, the day-to-day variability of most parameters was low, with coefficient of variation ranging from 1.8% to 10.7%. Microvascular parameters differed in individuals with ADPKD compared with controls, despite preserved eGFR. Interestingly, cortical vessel density and its derived perfusion correlated directly with eGFR, whereas tortuosity indices presented an inverse correlation. Conclusions Ultrasound localization microscopy provided a safe and reliable in vivo assessment of the human intrarenal microvasculature.

Abstract Introduction Peyronie’s Disease (PD) is a fibrotic disorder of the tunica albuginea leading to penile curvature, pain, and sexual dysfunction. Intralesional therapies are a non-surgical treatment strategy aiming to modulate fibrosis and reduce deformity. Intralesional verapamil, a calcium channel blocker, has been used to inhibit fibroblast proliferation and collagen synthesis, but its clinical effectiveness remains controversial. Despite widespread use, existing literature is heterogeneous in design and outcome measures. Objective We aimed to systematically review and summarize the evidence from prospective clinical trials assessing the efficacy and safety of intralesional verapamil. Methods Data extraction was carried out independently by the presenting author and a board-certified librarian. We conducted a structured literature search using PubMed, Scopus, and Web of Science for prospective trials from 2015-2025 evaluating intralesional verapamil use in patients with PD. Inclusion criteria were: [1] prospective study design, [2] adult patients with a clinical diagnosis of PD, [3] use of intralesional verapamil as monotherapy or in combination, and [4] reported outcomes on penile curvature, and sexual function. Data extraction included study design, sample size, outcome measures, and follow-up. Risk of bias and quality assessment was performed using the Cochrane Risk of Bias 2.0 scale. Results A total of 199 studies were screened. A total of 6 prospective trials were selected after excluding trials who were not compliant with our criteria, reporting on 754 patients with 1 to 24 months follow-up. RCTs evaluated were of moderate quality with a low risk of bias, but 3/6 had a high risk of bias. Across randomized trials, verapamil monotherapy resulted in modest improvements in curvature (range mean change in penile curvature -2° to -16°). In contrast, combination therapies or alternative injectables such as collagenase (CCH) and tiocolchicine showed greater curvature improvements (–28° in Chung; ~10° in Toscano). In non-randomized studies comparative studies, verapamil (Ver) demonstrated consistent improvement in penile curvature (~4.5° reduction [Cocci 22]), with sustained results at 24 months. Nevertheless, in these studies HA was superior to Verapamil (~9.5° reduction). Erectile function, measured by IIEF-5, improved modestly across studies, with the greatest gains observed in combination therapy arms or HA groups. Conclusions Intralesional verapamil offers modest improvements in penile curvature and erectile function in Peyronie’s Disease but appears less effective than alternative agents such as hyaluronic acid, collagenase, or combination therapies. Heterogeneity in trial design and moderate risk of bias limit definitive conclusions. Further high-quality, blinded randomized trials are needed to establish optimal intralesional treatment strategies. Disclosure No

Abstract Introduction Down-sized inflatable penile prosthesis (IPP) cylinders are useful in cases with substantial penile fibrosis such as history of priapism or prior infection. Although down-sized IPP cylinders have a clear role in fibrotic corpora, their role in primary implant cases is unclear. Objective To identify factors associated with the use of down-sized cylinders in men undergoing primary IPP placement. Methods We conducted a retrospective analysis of 4,389 patients undergoing primary IPP placement from our multicenter database. Patients were categorized by down-sized (AMS 700 CXR, Coloplast NB) vs. standard sized cylinders (AMS 700 CX, AMS 700 LGX, Coloplast Titan). We collected demographic, intraoperative, and postoperative data. A multivariable analysis was performed to identify factors associated with use of down-sized cylinders Results A total of 4,389 men undergoing primary inflatable penile prosthesis (IPP) placement were included in the study, with a mean age of 62.3 ± 10.3 years. Among them, 1,485 (34%) had diabetes and 2,131 (49%) had hypertension. A total of 406 down-sized cylinders and 3,983 standard cylinders were implanted. Non-infectious complications occurred more frequently with the use of down-sized cylinders (odds ratio [OR] = 2.0, p &lt; 0.001). Specifically, non-infectious complications were reported in 44 out of 185 men (23.8%) with down-sized cylinders, compared to 192 out of 2,727 men (10.9%) with standard cylinders. Cylinder complications constituted 13.9% and 13.0% of the non-infectious complications in the down-sized and standard cylinder groups, respectively. Conclusions The rate of cylinder-related complications were similar between the down-sized cylinder and standard cylinder groups. Urologists should be prepared to place down-sized cylinders during IPP placement in diabetic men and in men requiring serial dilation due to fibrosis. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Intuitive Surgical, Fidelis, Urofill, Boston Scientific, Coloplast, Cynosure, Antares Pharma, Clarus Pharmaceuticals, Acerus Pharma; Endo Pharma.

Abstract Introduction The Penuma ® implant is a medical-grade silicone implant used in the cosmetic augmentation of the penis. Reportedly 3-11% of devices are being removed over time for reasons such as infection, pain, and dissatisfaction with the device. Objective Despite this, there remains a paucity of information demonstrating safe removal of this device. In this video, we outline intra-operative considerations to promote the safe removal of the Penuma device. Methods Sharp and blunt dissection to the level of the capsule is performed via the original incision line. The proximal end of the device is identified via sharp dissection through the capsule. The penis is then everted to expose the distal attachments of the implant, which are subsequently dissected. Superficial and sharp dissection to remove the mesh is performed. We avoid the use of cautery to prevent damage to the nearby neurovascular bundle. Once the mesh and all permanent material is removed, haemostasias is achieved. A drain may be placed. The incision is irrigated and closed in a multi-layer fashion. Results Here we demonstrate a safe and feasible method to surgically remove the Penuma (Himplant) device. Conclusions In conclusion, the Penuma ® implant is a medical-grade silicone implant used in the cosmetic augmentation of the penis that can be safely removed surgically in the event of infection, pain, or dissatisfaction with the device. Disclosure No

Abstract Introduction Traditional vasectomy reversals are performed in the operating room under general anesthesia, which poses logistical, financial, and clinical drawbacks. A shift toward office-based vasectomy reversal under local anesthesia may address these limitations. Objective To demonstrate the feasibility and technical approach of performing microsurgical vasovasostomy in an office-based setting using local anesthesia, without compromising patient comfort or surgical technique. Methods In this video we outline a step-by-step microsurgical vasovasostomy in an office-based setting. Results Office-based vasovasostomy is technically feasible in the office-based setting. Conclusions Office-based microsurgical vasovasostomy is safe, feasible, and well tolerated in the office-based setting, offering multiple patient- and provider-centered advantages. Disclosure No