OBJECTIVE To identify differences in neuroinflammatory gene expression in individuals with chronic orchialgia (CO) compared to asymptomatic controls. METHODS Vas deferens, spermatic cord fascia, blood, and urine were collected from 9 men with CO at time of microscopic spermatic cord denervation and 7 asymptomatic controls at time of vasectomy. RNA was isolated and analyzed with the NanoString Human Neuroinflammation panel. Data were normalized, gene expression fold changes and enriched pathways relative to asymptomatic controls were determined. Gene expression was considered significantly different if there was a >2-fold change and P-value <.05 relative to controls. RESULTS Mean patient age was 51 years and median symptom duration 12 months. There were 26 genes with significantly differential expression in vas deferens. cFos, a marker of nociceptive pain, had the greatest difference (30.2-fold change, P <.000001). Enriched pathways in vas deferens included nerve function, matrix remodeling, and innate immune responses. In fascia, cFos also had the greatest differential expression (38-fold, P = .000002), followed by S100A12 (11-fold, inducer of innate immune response). Enriched pathways in fascia included nerve function and inflammation. In blood, there were no differentially expressed genes, and in urine there were 95 differentially expressed genes. CONCLUSION Men with CO have a diverse set of neuroinflammatory genes with differential expression in tissue and urine relative to healthy controls. These findings confirm pathologic changes in tissue targeted by denervation surgery, and suggest molecular changes in neuropathic pain that could lead to biomarker (c) 2022 Elsevier Inc.

BACKGROUND:Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has diverse clinical phenotypes and its etiology is multifactorial. Studies to date of gene expression in humans have been limited to small numbers of target genes. NanoString can simultaneously measure hundreds of genes. We wished to study gene expression in blood and urine of CP/CPPS patients compared to controls for neuroinflammatory genes and characterize the results by patient phenotype.METHODS:Blood and urine were collected from 10 men with CP/CPPS and 7 asymptomatic controls. RNA was isolated from urine pellets using Qiagen RNeasy kits. Whole blood was collected and RNA isolated. 100 ng of RNA was used for gene expression analysis with the 770-gene NanoString Human Neuroinflammation gene panel. Data was imported into Rosalind (OnRamp Bioinformatics) for normalization, calculation of fold-changes and P values, and identification of enriched pathways. Gene expression was considered significantly different if there was a greater than 1.5× change compared to controls and corrected P was <0.05.RESULTS:Mean patient age was 42.2 years, median symptom duration was 15.5 months, median UPOINT domains was 3 and mean total National Institute of Health-Chronic Prostatitis Symptom Index Score was 28.8. In blood, there were 5 genes with significantly different expression to controls, the largest differences found in FOS1 (neuropathic pain control), PROS1 (blood clotting) and DDX58 (antiviral innate immunity). Gene set analysis showed differences in inflammation, angiogenesis and cytokine signaling. In urine there were 48 genes with significantly different expression including SLAMF8 (lymphocyte activation) and LAIR1 (inhibits B and T cell function). Gene set analysis showed differences in carbohydrate metabolism, neurons and neurotransmission, adaptive immunity and inflammatory signaling. Subgroup analysis by UPOINT domain showed unique gene expression in the Organ Specific and Neurologic/Systemic domains in both blood and urine for neurogenic pain and cytokine signaling associated genes.CONCLUSIONS:Men with CP/CPPS have a diverse set of neuroinflammatory genes with differential expression compared to controls. Clinical phenotypes have distinct patterns of gene expression. These findings could lead to novel biomarker development, emphasize the importance of multimodal therapy targeting diverse pathways and further validate the biologic basic of clinical phenotyping.

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Prostate & Genitalia (MP35)1 Sep 2021MP35-13 NEUROINFLAMMATORY GENE EXPRESSION IN CHRONIC PROSTATITIS/CHRONIC PELVIC PAIN SYNDROME (CP/CPPS): INSIGHTS INTO ETIOLOGY AND PHENOTYPE BIOLOGY Johnathan Doolittle, Karen Keslar, Paige Gotwald, Sarah Vij, and Daniel Shoskes Johnathan DoolittleJohnathan Doolittle More articles by this author , Karen KeslarKaren Keslar More articles by this author , Paige GotwaldPaige Gotwald More articles by this author , Sarah VijSarah Vij More articles by this author , and Daniel ShoskesDaniel Shoskes More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002044.13AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: CP/CPPS has diverse clinical phenotypes and its etiology is multifactorial with inflammatory, immune and neuromuscular pathways implicated. Studies to date of gene expression in humans have been limited to small numbers of target genes. NanoString uses barcoded probes to simultaneously measure hundreds of genes. We wished to study gene expression in blood and urine of CP/CPPS patients compared to controls for a broad variety of neuroinflammatory genes and characterize the results by patient phenotype. METHODS: Blood and urine were collected from 10 men with CP/CPPS and 7 asymptomatic controls. RNA was isolated from urine pellets using Qiagen RNeasy kits. Whole blood was collected and RNA isolated using the Tempus Spin RNA isolation kit. 100 ng of RNA was used for gene expression analysis with the 770-gene NanoString Human Neuroinflammation gene panel. Data was imported into Rosalind (OnRamp Bioinformatics) for normalization, calculation of fold-changes and p values, and identification of enriched pathways. Gene expression was considered significantly different if there was a greater than 1.5x change compared to controls and corrected p was <0.05. RESULTS: Mean patient age was 42.2 years, median symptom duration was 15.5 months, median UPOINT domains was 3 and mean total NIH-CPSI score was 28.8. In blood, there were 5 genes with significantly different expression to controls, the largest differences found in FOS1 (neuropathic pain control), PROS1 (blood clotting and found in prostate cancer cells) and DDX58 (antiviral innate immunity). Gene set analysis showed differences in inflammation, angiogenesis and cytokine signaling. In urine there were 48 genes with significantly different expression including SLAMF8 (CD2 mediated lymphocyte activation) and LAIR1 (inhibits B and T cell function). Gene set analysis showed differences in Wnt (adaptive immune response in neuropathic pain), microglial function and inflammatory signaling. Subgroup analysis by UPOINT domain showed unique gene expression in the Psychosocial, Organ Specific and Neurologic/Systemic domains in both blood and urine for neurogenic pain and cytokine signaling associated genes. CONCLUSIONS: Men with CP/CPPS have a diverse set of neuroinflammatory genes with differential expression compared to controls including some never before implicated in the etiology of this condition. Furthermore, clinical phenotypes have distinct patterns of gene expression. These findings could lead to novel biomarker development, novel treatment options and further validates the biologic basic of clinical phenotyping. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e632-e632 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Johnathan Doolittle More articles by this author Karen Keslar More articles by this author Paige Gotwald More articles by this author Sarah Vij More articles by this author Daniel Shoskes More articles by this author Expand All Advertisement Loading ...

No AccessJournal of UrologyJU Forum1 Sep 2021Impact of COVID-19 on Prevention of Urinary Stones with Hydration (PUSH) Study: Challenges and Opportunities for Future Trials The Urinary Stone Disease Research Network (USDRN) Investigators The Urinary Stone Disease Research Network (USDRN) Investigators View All Author Informationhttps://doi.org/10.1097/JU.0000000000001833AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail References 1. : Clinical trials in urology: predictors of successes and failures. J Urol 2020; 204: 805. Link, Google Scholar 2. : Prevention of Urinary Stones with Hydration (PUSH): design and rationale of a clinical trial. Am J Kidney Dis 2021; 77: 898. Google Scholar 3. : An observational study of the association of video- versus text-based informed consent with multicenter trial enrollment: lessons from the PALM study (Patient and Provider Assessment of Lipid Management). Circ Cardiovasc Qual Outcomes 2018; 11: e004675. Google Scholar This research was supported by the National Institutes of Health/NIDDK, as follows: U01DK110961 (UPenn/CHOP—PP Reese, GE Tasian), U01KD110986 (Washington University in St. Louis—AC Desai, HH Lai), U01DK110994 (UT Southwestern—NM Maalouf), U01DK110954 (University of Washington—JD Harper, H Wessells), and U01DK110988 (Duke University—CD Scales, HR Al-Khalidi). Financial interest and/or other relationship with Allena Pharmaceuticals (CD Scales). © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue 3September 2021Page: 502-504 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.AcknowledgmentsUrinary Stone Disease Research Network: The following individuals were instrumental in the planning and conduct of the PUSH study at each of the participating institutions. Clinical Centers: University of Pennsylvania/Children’s Hospital of Pennsylvania, Philadelphia, Pennsylvania: Principal Investigator: Peter P. Reese, MD, MSCE, Gregory E. Tasian, MD, MSCE; Co-Investigators: Sandra Amaral, MD, MHS, Janet Audrain-McGovern, PhD; Study Coordinators: Emily Funsten, Brittney Henderson, Kristen Koepsell, Adam Mussell. University of Texas Southwestern Medical Center, Dallas, Texas: Principal Investigator: Naim M. Maalouf, MD; Co-Investigators: Jodi A. Antonelli, MD, Linda A. Baker, MD, Margaret S. Pearle, MD, PhD, Lakshmi Ananthakrishnan, MD; Study Coordinators: Joyce Obiaro, Cynthia Rangel, Martinez Hill, Madeline Worsham. University of Washington, Seattle, Washington: Principal Investigator: Jonathan D. Harper, MD, Hunter Wessells, MD; Co-Investigators: Fionnuala Cormack, MD, Mathew Sorensen, MD, Karyn Yonekawa, MD; Study Coordinators: Holly Covert, Tristan Baxter, Elsa Ayala. Washington University in St. Louis, St. Louis, Missouri: Principal Investigator: Alana C. Desai, MD, H. Henry Lai, MD; Co-Investigators: Vincent Mellnick, MD, Douglas Coplen, MD; Study Coordinators: Juanita Taylor, Aleksandra Klim, Deborah Ksiazek. Recruiting Centers: Cleveland Clinic Foundation, Cleveland, Ohio: Principal Investigator: Sri Sivalingam, MD, MSc, FRCSC; Co-Investigators: Katherine Dell, MD, Juan Calle, MD; Study Coordinators: Paige Gotwald, Marina Markovic. Mayo Clinic Foundation, Rochester, Minnesota: Principal Investigator: John Lieske, MD; Co-Investigators: Andrew Rule, MD, Stephen Erickson, MD, Aaron Potrezke, MD, Andrea Ferrero, PhD, David Sas, DO; Study Coordinators: Angela Waits, Courtney Lenort. Scientific Data Research Center: Duke Clinical Research Institute, Duke University, Durham, North Carolina: Principal Investigator: Charles D. Scales, Jr., MD, MSHS, Hussein R. Al-Khalidi, PhD; Co-Investigators: Kevin Weinfurt, PhD, Hayden Bosworth, MD; Statistician: Honqiu Yang, PhD; Project Lead: Laura Johnson; Lead CRA: Sharon Settles; CRA: Angela Venetta; Data Manager: Omar Thompson. National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK): Project Scientist: Ziya Kirkali, MD; Program Official: Christopher Mullins, PhD. Data Safety and Monitoring Board: John Denstedt, MD (Chair), Dean G. Assimos, MD, Uri Alon, MD, Scott Cohen, MD, Michael A Freeman, MD, Rebecca A Krukowski, PhD, Jeannette Lee, PhD, Eric Taylor, MD, Jennifer Temple, PhD, Christopher H Schmid, PhD. Past members: Gary C. Curhan, MD, ScD, David S. Goldfarb, MD, Manoj Monga, MD, FACS, Andrew Rule, MD, Marshall Stoller, MD.Metrics Author Information The Urinary Stone Disease Research Network (USDRN) Investigators More articles by this author Expand All This research was supported by the National Institutes of Health/NIDDK, as follows: U01DK110961 (UPenn/CHOP—PP Reese, GE Tasian), U01KD110986 (Washington University in St. Louis—AC Desai, HH Lai), U01DK110994 (UT Southwestern—NM Maalouf), U01DK110954 (University of Washington—JD Harper, H Wessells), and U01DK110988 (Duke University—CD Scales, HR Al-Khalidi). Financial interest and/or other relationship with Allena Pharmaceuticals (CD Scales). Advertisement PDF downloadLoading ...

Background: We sought to study differential neuroinflammatory gene expression in men with interstitial cystitis (IC) with Hunner lesions compared with asymptomatic controls using NanoString, which uses barcoded probes to measure hundreds of genes. IC is a heterogenous condition lacking reliable biomarkers, and a subset of patients exhibits Hunner lesions, implicating the bladder as an inflammatory pain generator. Methods: Blood, urine, and bladder biopsies were collected from 6 men with IC and Hunner lesions. 7 asymptomatic controls had blood and urine collected and 2 benign bladder biopsies were obtained from our tissue bank. RNA was isolated and analyzed with NanoString Human Neuroinflammation panel. Gene expression was considered significant if there was a >1.5-fold change and adjusted P value <0.05 compared with controls. Results: Mean patient age was 61.5 years with 8 years median symptom duration. In bladder tissue, while many cytokine and chemokine genes had higher expression as expected (e.g., TNF, CXCL10), other significant genes included TRPA1 (1098-fold increased, expressed in pain sensing neurons) and TNFRSF17 (735-fold, B-cell related). In urine, there was 114-fold increase in S1PR4, which mediates pain via TRP-dependent pathways. A patient on cyclosporine had lower inflammatory gene expression levels relative to other IC patients, but no difference in TRPA1. Conclusions: Men with IC and Hunner lesions have a diverse set of neuroinflammatory genes with differential expression compared to controls. We identified genes linked to neuropathic pain through the TRP pathway and this expression was not reduced by cyclosporine. These findings open a new direction for biomarker and therapeutic discovery.

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Prostate & Genitalia (MP58)1 Apr 2020MP58-05 DETECTION OF BACTERIA BY NEXT GENERATION SEQUENCING IN MEN WITH CHRONIC PROSTATITIS/CHRONIC PELVIC PAIN SYNDROME: INCIDENCE, CORRELATION TO CONVENTIONAL CULTURE AND IMPACT ON SYMPTOMS Nicholas Farber*, Paige Gotwald, and Daniel Shoskes Nicholas Farber*Nicholas Farber* More articles by this author , Paige GotwaldPaige Gotwald More articles by this author , and Daniel ShoskesDaniel Shoskes More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000927.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Chronic prostatitis/Chronic pelvic pain syndrome (CPPS) is a syndrome that shares clinical features with urinary infections and a certain subset of patients improve with antibiotics. However, traditional cultures of urine and expressed prostatic secretions often fail to identify an organism. Next-generation sequencing (NGS) analyzes microbial DNA and can identify organisms that fail to grow in traditional cultures. We sought to compare traditional cultures with NGS in men with CPPS and examine the impact on symptoms and treatment response. METHODS: 25 men with a clinical diagnosis of CPPS underwent both traditional cultures and NGS (MicroGen Dx) of urine and expressed prostatic secretions (EPS). NIH-Chronic Prostatitis Symptom Index (NIH-CPSI) and UPOINT domains were evaluated. Patients with negative traditional cultures and positive NGS were offered antibiotic therapy. RESULTS: Urine cultures were negative in all CPPS patients, while 8% (2/25) had a positive EPS culture. NGS identified these organisms in one patient and failed to detect it in the other. In culture negative patients, NGS identified at least one organism in EPS in 70% (16/23), though only 30% (7/23) were for established uropathogens. Patients with positive and negative NGS had similar mean NIH-CPSI scores of 23.3 +/- 6.8 and 20.5 +/- 6.9, respectively (p=0.44). Organisms of questionable pathogenicity in EPS found by NGS included vaginal flora (Prevotella spp., Sneathia amnii), anaerobes, and fungi. In the subset of 6/25 (24%) men with systemic symptoms suggestive of infection, all had negative EPS cultures but NGS identifying a uropathogen in 50% (3/6). 4 were treated with antibiotics based on the sensitivity gene panel but only 1 patient (25%) resolved their symptoms. CONCLUSIONS: NGS detected a variety of microorganisms not found by conventional culture in urine and EPS. In men with systemic symptoms suggestive of infection, NGS found a pathogen missed by culture in 50%. The presence of vaginal flora, anaerobes, and yeast is novel but the clinical significance is unclear and the value of prolonged antimicrobial treatment of these organisms is not proven. Based on our data, NGS may help identify pathogenic organisms in men with CPPS who have symptoms suggestive of true infection, however men should still be offered multimodal therapy based on their clinical phenotype to maximize the chance for symptom resolution. Source of Funding: None © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e872-e873 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nicholas Farber* More articles by this author Paige Gotwald More articles by this author Daniel Shoskes More articles by this author Expand All Advertisement PDF downloadLoading ...

OBJECTIVE To compare standard cultures and next-generation sequencing (NGS) in men with chronic prostatitis/chronic pelvic pain syndrome (CPPS). CPPS shares clinical features with urinary tract infections, but bacteria are seldom found. NGS is more sensitive than standard cultures. MATERIALS AND METHODS Men diagnosed with CPPS (National Institute of Health Category III) underwent traditional cultures and NGS of their urine and expressed prostatic secretions (EPS). Characteristics between groups were compared statistically. RESULTS Thirty-one men with CPPS were included (mean age 44.5). All standard urine cultures were negative, and 3 EPS cultures were positive. Seventy-eight unique microbes were detected with NGS, including uropathogens in 10 of the men. There were no bacteria identified by NGS in EPS that were not also found in the urine. Men with positive NGS did not differ from those without in age, symptom severity or phenotype. Men with typical urinary tract infection symptoms (eg, dysuria, chills) were more likely to have uropathogens detected on NGS relative to men without such symptoms. Nine patients were prescribed antibiotics based on their NGS findings, but only 1 exhibited symptom improvement (11%). CONCLUSION NGS commonly identified bacteria in CPPS patients, but these did not localize to the prostate. NGS positivity did not correlate with symptom severity and antibiotic therapy was seldom effective. NGS detected uropathogens more frequently in those with clinical symptoms suggestive of urinary tract infection. Clinical trials are needed to examine the utility of NGS-guided antibiotics in this subpopulation. (C) 2020 Elsevier Inc.