Grant R01 Research Articles

MP14-06 URETERAL STENTS ASSOCIATED WITH LOWER COMPLICATION RATES VERSUS PERCUTANEOUS NEPHROSTOMY TUBES IN THE TREATMENT OF MATERNAL NEPHROLITHIASIS

You have accessJournal of UrologyCME1 May 2022MP14-06 URETERAL STENTS ASSOCIATED WITH LOWER COMPLICATION RATES VERSUS PERCUTANEOUS NEPHROSTOMY TUBES IN THE TREATMENT OF MATERNAL NEPHROLITHIASIS Matthew Mason, Sirpi Nackeeran, Marisa Carino Mason, Taylor Kohn, and Ranjith Ramasamy Matthew MasonMatthew Mason More articles by this author , Sirpi NackeeranSirpi Nackeeran More articles by this author , Marisa Carino MasonMarisa Carino Mason More articles by this author , Taylor KohnTaylor Kohn More articles by this author , and Ranjith RamasamyRanjith Ramasamy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002543.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: In complicated cases of maternal nephrolithiasis, placement of a percutaneous nephrostomy tube (PCN) or ureteral stent for temporary drainage is recommended. Although there have been small institutional and case series studies to compare each intervention’s complication rates, there is a paucity of literature doing so using a large database. We therefore investigated complication rates in the use of PCNs versus ureteral stents in the treatment of maternal nephrolithiasis using a multi-institutional, national claims database. METHODS: We queried the TriNetX network database: a global health research network consisting of 65 million patients in 44 healthcare organizations. Pregnant women (ICD-10 Z34, O09) with a history of complicated maternal nephrolithiasis (N20-23) who underwent a PCN (CPT 50432) or ureteral stent (52332) placement within 6 months of pregnancy diagnosis were evaluated for incidence of any additional inpatient admission, premature labor or delivery (O42, O60), exchange procedure (52332, 52310, 52315, 50387, 50432, 50435), new UTI or pyelonephritis (N39.0, O23, N10), or refractory nephrolithiasis (N20-23) within 1 week to 6 months of their initial procedure. We controlled for the following potentially confounding variables through propensity-score matching: age, race, ethnicity, and previous pyelonephritis or sepsis (N10, O23, A41). RESULTS: We identified 883 pregnant women who underwent ureteral stent placement and 136 who underwent PCN. Within 6 months of their initial procedure and following propensity score matching, significantly less women with a ureteral stent experienced a new UTI or pyelonephritis (aOR 0.37; 95% CI 0.15-0.94; p=0.0351) compared to those who underwent PCN placement. Ureteral stent placement was also associated with a significantly decreased rate of hospital admissions (aOR 0.37; 0.22-0.61; p<0.0001) and less need for repeat ureteral stent or PCN (aOR 0.47; 0.29-0.78; p=0.0028). There was no difference in rates of premature labor or delivery (aOR 0.83; 0.41-1.66; p=0.5936), or refractory nephrolithiasis (aOR 0.87; 0.42-1.80; p=0.1259). CONCLUSIONS: In maternal nephrolithiasis, ureteral stent placement was associated with a lower incidence of hospital admissions, exchange procedures, and new UTIs or pyelonephritis. Although further studies are needed to assess differences in temporality and severity of disease post-procedure, this study assists providers in weighing clinical risks and benefits in the treatment of complicated maternal nephrolithiasis. Source of Funding: This work was supported by National Institutes of Health Grant R01 DK130991 from the American Cancer Society to Ranjith Ramasamy © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e234 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Matthew Mason More articles by this author Sirpi Nackeeran More articles by this author Marisa Carino Mason More articles by this author Taylor Kohn More articles by this author Ranjith Ramasamy More articles by this author Expand All Advertisement PDF DownloadLoading ...

Investigating the role of inflammasome activation by dengue virus non-structural protein 1 during dengue infection.

Abstract Dengue virus (DENV), consisting of serotypes 1–4, is the most medically important flavivirus, causing an estimated 50–100 million dengue cases globally every year. Most symptomatic DENV infections present as an acute febrile illness; however, severe complications include vascular leakage leading to hemorrhage and hypovolemic shock. DENV non-structural protein 1 (NS1) is secreted from infected cells and has been implicated as a major driver of dengue pathogenesis, activating immune cells and acting directly on endothelial barriers, causing them to become pathologically hyperpermeable. While recent work has delved into the mechanisms behind the endothelial cell-intrinsic pathway of DENV NS1-induced vascular leak, much less is known about how DENV NS1 interacts with immune cells and what role this activation plays in DENV infection. We have discovered that DENV NS1 can trigger activation of the inflammasome, a family of cytosolic innate immune sensors that react to danger-associated molecular patterns. DENV NS1 induces the release of the pro-inflammatory cytokine IL-1β in human and murine monocytes in a caspase-1 dependent manner that is independent of NLRP3. Additionally, we found that caspase-1-deficient, but not NLRP3-deficient, mice are more susceptible to infection in a murine model of DENV infection. These results indicate that the inflammasome acts as a sensor of the DENV viral toxin NS1 and plays a protective role during DENV infections This work was supported by NIAID/NIH grant R01 AI24493 (E.H.)

MP10-01 FACTORS IN UROLOGY JOURNALS THAT DETERMINE FEMALE AUTHORSHIP REPRESENTATION: RETROSPECTIVE REVIEW FROM 2015 TO 2020

You have accessJournal of UrologyCME1 May 2022MP10-01 FACTORS IN UROLOGY JOURNALS THAT DETERMINE FEMALE AUTHORSHIP REPRESENTATION: RETROSPECTIVE REVIEW FROM 2015 TO 2020 Maria Camila Suarez, Daniel E. Nassau, Arjun Watane, Aayush Shah, and Ranjith Ramasamy Maria Camila SuarezMaria Camila Suarez More articles by this author , Daniel E. NassauDaniel E. Nassau More articles by this author , Arjun WataneArjun Watane More articles by this author , Aayush ShahAayush Shah More articles by this author , and Ranjith RamasamyRanjith Ramasamy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002532.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Proportional to their representation in urology, women remain scarce in leadership positions and high academic ranks. Although this gap is likely multifactorial, research productivity stands out as one of the main determinants, with female urologist exhibiting lower Hirsch index than their male counterparts. The reasons behind this gender-disparity in female authorship has not been elucidated. The aim of this study is to evaluate if the CiteScore (CS), scope (general vs subspecialty), gender-association among authors, and peer-review in the leading urology journals determine the representation of females as first and senior authors. METHODS: A computer script was designed using Python™ to search PubMed and record first and last authors of publications between 2015-2020, from the eighteen urology journals with the highest CS. At least one journal per subspecialty was included. Gender was determined by ©Gender-API (https://gender-api.com), and names with an accuracy >90% were included. Student t-test and ANOVA were implemented to compare means, Jonckheere Terpestra’s test to assess trends, Chi-square to establish proportions, and Fisher’s exact test to calculate odds ratios. A significant p-value was considered <0.05. RESULTS: A total of 37,414 publications and 65,828 authors were included in the study. Female authorship prevalence was 18.6%, and the proportion of female first authors was significantly greater than that of female senior authors (21% vs 15%, p <.001). Journals within the top third CS had the lowest proportion of female authors, while journals at the bottom third CS had the highest (p <.001). Overall, articles with female first authors were more likely to have a female senior author (OR: 2.72, CI: 2.53-2.92, p <.001), and 15 of the 18 journals showed a gender association. Analysis by scope showed that subspecialty journals had a larger proportion of manuscripts authored by female senior authors than general journals (p=0.046). When female authorship was compared by type of peer-review no significant difference was observed (p=0.09). Nonetheless, when only subspecialty journals were analyzed, double-blinded peer-reviews demonstrated a greater proportion of female first and last authors than single-blinded journals (p <.001). Over the study period, the proportion of females as last authors increased by 3.2% (p=0.045); no significant change in the proportion of females as first authors was observed (p=0.094). CONCLUSIONS: Our data suggests that the proportion of female authorship is determined by the journal’s CS, scope, peer-review, and gender association between first and senior authors. Comprehending the causes of authorship inequity should be a priority in order to create strategies for reducing gender-disparities in the recruitment and promotion of women in academia and leadership position in urology. Source of Funding: This work was supported by National Institutes of Health Grant R01 DK130991 and Clinician Scientist Development Grant from the American Cancer Society to RR. © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e146 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Maria Camila Suarez More articles by this author Daniel E. Nassau More articles by this author Arjun Watane More articles by this author Aayush Shah More articles by this author Ranjith Ramasamy More articles by this author Expand All Advertisement PDF DownloadLoading ...

Early post-vaccination gene signatures correlate with the magnitude and function of vaccine-induced HIV envelope-specific plasma antibodies in infant rhesus macaques

Abstract Systems vaccinology approaches are important tools in rational vaccine design. Our goal was to determine whether early innate immune responses to the vaccine prime in infant rhesus macaques, immunized with two different HIV envelope (Env) vaccine regimens, were associated with functional antibody responses in the memory phase. We compared plasma cytokine levels and molecular signatures of a 3M-052-SE adjuvanted HIV Env protein vaccine (n=10) to a regimen combining the adjuvanted HIV Env protein and MVA-HIV Env (n=10) at days (D) 0, 1, and 3 post the vaccine prime. Whole blood transcriptional profiling applying NanoString technology was employed to identify differentially expressed genes (DEG). Innate immune responses were correlated with vaccine-induced adaptive immune responses at weeks 14, 20, 32, and 34. The vaccine prime induced a rapid, but transient, increase in inflammatory plasma cytokines and changes in mRNA expression that peaked at D1. In the HIV protein group, we identified 31 DEG with increased mRNA levels, whereas a single, downregulated, DEG was identified in the MVA-Env plus Protein group. Day one signatures were positively correlated with week 14 Env-specific IgG responses, and, at week 34, with Env-specific follicular T helper cells and Env-specific antibody-dependent cytotoxicity function, but negatively correlated with Env-specific CD8+ T cell responses. A protein-protein interaction network confirmed that several of the DEG-encoded proteins have predicted interaction partners that are important for B cell activation. These results support the idea that vaccine-induced HIV-specific antibody and T cell responses can be optimized through the modulation of the vaccine prime. Supported by National Institutes of Health grants R01 DE028146 , P01 AI117915 , T32 5108303 , the Office of Research Infrastructure Programs/OD P510D011107 (CNPRC), and the Center for AIDS Research award P30AI050410

Cleavage of DNA and RNA by PLD3 and PLD4 limits autoinflammatory triggering by multiple sensors, including endolysosomal TLRs and a STING dependent sensing pathway.

Abstract We have studied the biochemistry and genetics of Phospholipase D3 (PLD3) and PLD4 because polymorphisms of these proteins have been associated with several important inflammatory and autoimmune diseases, including rheumatoid arthritis, systemic sclerosis, systemic lupus erythematosus and Alzheimer’s disease. We found that both PLD3 and PLD4 are 5’ exonucleases able to degrade both ssDNA and ssRNA. Deficiency in PLD3, PLD4 or both results in a variety of inflammatory defects, with the most serious being lethal hemophagocytic lymphohistiocytosis (HLH) in double knockout mice. Pld3−/−Pld4−/− mice represent a unique spontaneous model of HLH apparently independent of microbes, whereas Pld4−/− mice develop a range of phenotypes depending upon genetic background ranging from macrophage activation syndrome to lupus like disease. Evidence will be presented showing that the diseases in these mice result from autorecognition of nucleic acids and can be reversed by mutation of nucleic acid sensing pathways. PLD3 and PLD4 tend to limit TLR9 recognition but have a more complex effect on recognition by TLR7 and TLR8. Unc93b13d/3dPld3−/−Pld4−/− mice spontaneously signal through STING suggesting that PLD3 and PLD4 also may regulate cytoplasmic nucleic acid sensing. Supported by NIH grants R01 AI142945 and R56AG070775

PD16-02 ULTRASTRUCTURAL FINDINGS OF COVID-19 INFECTION IN THE PROSTATE

You have accessJournal of UrologyCME1 May 2022PD16-02 ULTRASTRUCTURAL FINDINGS OF COVID-19 INFECTION IN THE PROSTATE Rohit Reddy, Eliyahu Kresch, Natoli Farber, Parris Diaz, and Ranjith Ramasamy Rohit ReddyRohit Reddy More articles by this author , Eliyahu KreschEliyahu Kresch More articles by this author , Natoli FarberNatoli Farber More articles by this author , Parris DiazParris Diaz More articles by this author , and Ranjith RamasamyRanjith Ramasamy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002548.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: SARS-CoV-2 utilizes two integral membrane proteins ACE2 and TMPRSS2 for viral replication. It has been established TMPRS22 specifically is found in high concentrations throughout the prostate found to be linked to prostatic disease progression. This project examined the histopathological, ultrastructural, and immunofluorescent elements of prostatic tissue from men infected by SARS-CoV-2. METHODS: We evaluated prostate tissue in men with worsening lower urinary tract symptoms who underwent HoLEP procedure after SARS-CoV-2 infection. Biopsied tissue was visualized by transmission electron microscopy (TEM), immunofluorescence, and viral presence was confirmed by quantitative polymerase chain reaction (RT-PCR). RESULTS: Multiple coronavirus-like spiked viral particles ranging from 73.3mm to 109mm were visualized by TEM (Figure). Histochemical and immunofluorescence concurrently showed presence of distinct hyalinization, fibrosis, and presence of spike protein (Figure 2). RT-PCR confirmed the identity of the viral bodies as SARS-CoV-2 (Figure 3). CONCLUSIONS: This study provides evidence that SARS-CoV-2 not only enters prostatic tissue but may persist beyond initial infection period. In addition to establishing the persistence of SARS-CoV-2 particles in prostatic tissue, this report suggests the importance of discerning the relationships between COVID-19, lower urinary tract symptom severity, and prostatic hyperplasia. 2 Source of Funding: This work was supported by National Institutes of Health Grant R01 DK130991 and Clinician Scientist Development Grant from the American Cancer Society to RR © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e269 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rohit Reddy More articles by this author Eliyahu Kresch More articles by this author Natoli Farber More articles by this author Parris Diaz More articles by this author Ranjith Ramasamy More articles by this author Expand All Advertisement PDF DownloadLoading ...

PD09-12 IS THE SEMEN MICROBIOME PROFILE IN MEN WITH NON-OBSTRUCTIVE AZOOSPERMIA DIFFERENT FROM FERTILE CONTROLS?

You have accessJournal of UrologyCME1 May 2022PD09-12 IS THE SEMEN MICROBIOME PROFILE IN MEN WITH NON-OBSTRUCTIVE AZOOSPERMIA DIFFERENT FROM FERTILE CONTROLS? Joseph M. Israeli, Maria Camila Suarez, Daniel E. Nassau, Kevin Y. Chu, Emad Ibrahim, and Ranjith Ramasamy Joseph M. IsraeliJoseph M. Israeli More articles by this author , Maria Camila SuarezMaria Camila Suarez More articles by this author , Daniel E. NassauDaniel E. Nassau More articles by this author , Kevin Y. ChuKevin Y. Chu More articles by this author , Emad IbrahimEmad Ibrahim More articles by this author , and Ranjith RamasamyRanjith Ramasamy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002535.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Recent findings reveal a unique seminal microbiome, although the impacts of microorganism composition on fertility are uncertain. We hypothesized that there is a difference in the seminal microbiome between non-obstructive azoospermia (NOA) and fertile controls. To investigate, we compared semen samples from men with NOA and fertile controls – men with vasectomy. We compared alpha diversity, beta diversity, and differential frequencies/abundances of microbes. METHODS: This is a prospective study of male patients presenting for fertility evaluation at a single institution. History was recorded from chart review. NOA was diagnosed using clinical parameters (small testis size and FSH > 10 IU/mL). Single semen samples were collected at Urology office visit for semen analysis, and then sent for Next Generation Sequencing (NGS). NGS involves both quantitative PCR and 16s rRNA V1/V2 region comprehensive sequencing using Illumina MiSeq technology. Microorganism species and relative abundances were documented. Beta diversity between groups and differential species abundances were assessed using the Bray-Curtis Diversity Index and Independent Sample T-Tests respectively. RESULTS: Since August 2021, NOA patients (average age 38.1) and post-vasectomy patients (average age 36.3) were enrolled. Unique bacterial species numbered 20 (NOA), and 17 (vasectomy). The average number of species per patient was 5 (NOA) and 4.43 (vasectomy). The most frequent organisms in men with NOA and fertile controls were [C. Tuberculostearicum, F. Magna, and S. Epidermidis], and [E. Faecalis, S. Epidermidis, S. Hominis, and C. Tuberculostearicum] respectively. The most abundant organisms in men with NOA and fertile controls were [E. Faecalis, L. Iners, and E. Coli], and [E. Faecalis, S. Epidermidis, and C. Tuberculostearicum] respectively. Bacteria solely found in one group are displayed in Table 1. The Bray-Curtis value of 0.47 reveals a substantial amount of beta diversity between groups. CONCLUSIONS: The seminal microbiome in NOA is different in composition and abundance from that of fertile controls. This is the first study to compare the microbiome in cases of NOA vs fertile controls. Further study is needed to determine the clinical utility of these observations. Source of Funding: Investigator Initiated Grant Supported by MicrogenDx to RR. This work was supported by National Institutes of Health Grant R01 DK130991 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e181 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Joseph M. Israeli More articles by this author Maria Camila Suarez More articles by this author Daniel E. Nassau More articles by this author Kevin Y. Chu More articles by this author Emad Ibrahim More articles by this author Ranjith Ramasamy More articles by this author Expand All Advertisement PDF DownloadLoading ...

Enrichment of Epstein Barr Virus in patients with Multiple Sclerosis

Abstract Multiple sclerosis (MS) is an autoimmune disease of the central nervous system affecting more than 2 million people worldwide and has been linked with Epstein-Barr Virus (EBV) infection. Previous studies by the Kottyan and Weirauch labs show that the EBNA2 protein of both EBV type 1 and 2 binds unique MS risk loci potentially causing changes at the transcriptional level contributing to MS disease progress. We hypothesize that type 1 and type 2 EBV will be detectable in the MS patient samples, and we aim to quantify EBV enrichment and identify EBV type in MS patient-derived blood samples. Analysis by qPCR showed that 30% of the control population (n=10) and 66% of the MS patient population (n=41) had detectable levels of EBV whereas the MS patients had a significantly higher frequency of EBV presence (X2 test p=0.0389). Of the samples that tested positive for EBV, 66% showed presence of type 1 EBV while 33% were unknown without a significant difference between MS patients and control group (X2 test p&amp;gt;0.9999). Due to issues with the limit of detection, droplet digital PCR was assessed for higher sensitivity compared to qPCR, but was found to have a comparable limit of detection to qPCR. To overcome the challenge of detecting low copy number viruses, we look toward targeted DNA capture and next generation sequencing to improve the sensitivity of EBV detection and specificity of EBV typing. Understanding the distribution of EBV types among MS patients would elucidate the precise role of EBV in MS development, potentially leading to improved treatment and prevention methods. Supported by grants from NIH grants R01 HG010730, R01 NS099068, R01 GM055479, and U01 AI130830 to MTW; R01 DK107502, R01 AI148276, U19 AI070235, U01 HG011172, and P30 AR070549 to LCK; R01 AR073228, R01 AI024717, and CCHMC ARC Award 53632 to MTW and LCK.

Mettl14-dependent m6A modification controls iNKT cells development and function

Abstract N6-Methyladenosine (m6A), the most common form of RNA modification, controls CD4+ T cell homeostasis by targeting the IL-7/STAT5/SOCS signaling pathways. The role of m6A modification in unconventional T cell development remains unknown. Using mice with T cell-specific deletion of RNA methyltransferase METTL14 (T-Mettl14 −/−), we demonstrated that m6A modification was indispensable for iNKT cell homeostasis. Loss of METTL14-dependent m6A modification led to the upregulation of p53-mediated apoptosis in double positive thymocytes, which in turn decreased distal Va-Ja gene rearrangements, including the invariant Va14-Ja18 TCR, resulting in a drastic reduction of iNKT numbers in the thymus and periphery. Residual T-Mettl14−/− iNKT cells exhibited increased apoptosis, impaired maturation, and decreased responsiveness to IL-2/IL-15 and TCR stimulation. Furthermore, METTL14 knockdown in mature iNKT cells diminished their cytokine production, correlating with increased Cish expression and decreased TCR signaling. Collectively, our study highlights a critical role for METTL14-dependent-m6A modification in iNKT cell development and function. This work was supported by NIH grants R01 AI41083 and R01 AI057460 to C.-R.W.

Inverse-agonist (SMM-189) suppresses colitis by inducing endogenous cannabinoids and attenuating Th17, neutrophils, natural killer cells

Abstract The cause of inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), are multi-factorial and include activated immune cells, chronic inflammation, genetics, and environmental exposure. It is well known that endocannabinoids mediate protection against intestinal inflammation and other autoimmune diseases. However, the effect of cannabinoid receptors induction by inverse-agonist during experimental colitis has not been investigated. Here, we investigate the activation of the cannabinoid receptor-2 (CB2) by SMM-189, a potent CB2 inverse agonist, on the inhibition of dextran sodium sulfate (DSS) induced colitis. In the present study, we found that SMM-189 effectively attenuated the overall clinical score, reversed colitis-associated pathogenesis, increased body weight and colon length while increasing the expression of CB2 receptors and protein kinase A (PKA) in the colon lamina propria (LPs). We also measured a decrease in the percentage and number of Th17 cells in the spleen, mesenteric lymph nodes (MLNs), and LPs treated with SMM-189 compared to DSS controls. Similarly, the percentage and number of natural killer T (NKT) cells and neutrophils are decreased after SMM-189 treatment. SMM-189 also induces the myeloid-derived suppressor cells (MDSCs) during chronic colitis progression as compared to DSS alone. These findings suggest that SMM-189 ameliorates experimental colitis by inducing CB2 receptors and PKA in the LPs, MDSCs and reduces Th17, NKT cells, and neutrophils from the spleen and LPs. Taken together, these data support the idea that the CB2 inverse agonists, such as SMM-189, may be developed as a novel therapeutic target for IBD. Supported by NIH grant R01 AI140405

Increased Pain Sensitivity Is Associated with Reduced REM Sleep in Females with Temporomandibular Joint Disorder (TMD)

Previous evidence demonstrates that the incidence of chronic sleep disturbance is high in patients with temporomandibular joint disorder (TMD). However, there has been little examination of whether heightened daytime pain sensitivity in this population is associated with alter sleep architecture at night. We aimed to test the extent to which daytime pain sensitivity, measured using quantitative sensory testing (QST) is associated with objective markers of sleep physiology, including sleep architecture and power spectral density bands, in patients with TMD. Female participants (n=149) with insomnia and TMD were recruited as part of a randomized-controlled trial. Pre-randomization, we performed a quantitative sensory testing (QST) battery to measure General Pain Sensitivity, a Central Sensitization Index, and Masseter Pressure Pain Threshold, and performed polysomnography (PSG) that night to calculate sleep architecture parameters (N1%, N2%, SWS%, REM%) and derive relative power in the Delta (0.5-3.5 Hz), Theta (3.5-8.0 Hz), Alpha (8.0-15.0 Hz), and Beta (15-30.0 Hz) spectral bands during sleep. We fit linear regression models with sleep parameters as the outcome, and pain sensitivity as the predictor, controlling for age, depressive symptom severity, and race. Higher Central Sensitization (B = -2.863, P = <0.01) and General Pain Sensitivity Indices (B = -3.069, P = <0.01) were significantly associated with lower REM% both before after controlling for covariates, however masseter pressure pain threshold was not significantly associated with REM% (B = -0.0259, P = 0.0743). Pain sensitivity measures were not significantly associated with relative power in any of the spectral bands, or with any other sleep architectural stages. Heightened pain sensitivity, measured using QST is associated with objective features of sleep architecture in TMD patients, potentially due to heightened pain interfering with sleep. These findings may contribute to understanding of physiological mechanisms through which acute and chronic pain result in sleep disruption. Grant support from NIH Grant R01 DE019731. Previous evidence demonstrates that the incidence of chronic sleep disturbance is high in patients with temporomandibular joint disorder (TMD). However, there has been little examination of whether heightened daytime pain sensitivity in this population is associated with alter sleep architecture at night. We aimed to test the extent to which daytime pain sensitivity, measured using quantitative sensory testing (QST) is associated with objective markers of sleep physiology, including sleep architecture and power spectral density bands, in patients with TMD. Female participants (n=149) with insomnia and TMD were recruited as part of a randomized-controlled trial. Pre-randomization, we performed a quantitative sensory testing (QST) battery to measure General Pain Sensitivity, a Central Sensitization Index, and Masseter Pressure Pain Threshold, and performed polysomnography (PSG) that night to calculate sleep architecture parameters (N1%, N2%, SWS%, REM%) and derive relative power in the Delta (0.5-3.5 Hz), Theta (3.5-8.0 Hz), Alpha (8.0-15.0 Hz), and Beta (15-30.0 Hz) spectral bands during sleep. We fit linear regression models with sleep parameters as the outcome, and pain sensitivity as the predictor, controlling for age, depressive symptom severity, and race. Higher Central Sensitization (B = -2.863, P = <0.01) and General Pain Sensitivity Indices (B = -3.069, P = <0.01) were significantly associated with lower REM% both before after controlling for covariates, however masseter pressure pain threshold was not significantly associated with REM% (B = -0.0259, P = 0.0743). Pain sensitivity measures were not significantly associated with relative power in any of the spectral bands, or with any other sleep architectural stages. Heightened pain sensitivity, measured using QST is associated with objective features of sleep architecture in TMD patients, potentially due to heightened pain interfering with sleep. These findings may contribute to understanding of physiological mechanisms through which acute and chronic pain result in sleep disruption. Grant support from NIH Grant R01 DE019731.

MP34-05 IMPACT OF NASAL, INTRAMUSCULAR, AND SUBCUTANEOUS TESTOSTERONE PELLETS ON INTRATESTICULAR TESTOSTERONE: EVALUATION OF DATA FROM TWO RANDOMIZED CLINICAL TRIALS

You have accessJournal of UrologyCME1 May 2022MP34-05 IMPACT OF NASAL, INTRAMUSCULAR, AND SUBCUTANEOUS TESTOSTERONE PELLETS ON INTRATESTICULAR TESTOSTERONE: EVALUATION OF DATA FROM TWO RANDOMIZED CLINICAL TRIALS Parris Diaz, Rohit Reddy, Ruben Blachman-Braun, Alexandra Dullea, Isaac J. Zucker, Daniel C. Gonzalez, Eliyahu Kresch, and Ranjith Ramasamy Parris DiazParris Diaz More articles by this author , Rohit ReddyRohit Reddy More articles by this author , Ruben Blachman-BraunRuben Blachman-Braun More articles by this author , Alexandra DulleaAlexandra Dullea More articles by this author , Isaac J. ZuckerIsaac J. Zucker More articles by this author , Daniel C. GonzalezDaniel C. Gonzalez More articles by this author , Eliyahu KreschEliyahu Kresch More articles by this author , and Ranjith RamasamyRanjith Ramasamy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002588.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Testosterone replacement therapy (TRT) can potentially cause decreased spermatogenesis and subsequent infertility due to decrease in intratesticular testosterone (ITT) production. Serum 17-hydroxyprogesterone (17-OHP) is a reliable surrogate for ITT that is critical for preserving male fertility. We hypothesized that long-acting T formulations will decrease ITT compared to short-acting formulations. We evaluated data from two open-label, randomized, two-arm clinical trials amongst different testosterone therapies with subcutaneous T pellets (TP), Intranasal Testosterone (NT) or Intramuscular Testosterone cypionate (TC). METHODS: A total of 75 symptomatic hypogonadal men (2 serum T <300 ng/dL drawn before 10AM) were randomized into open-label randomized clinical trials. Eligible subjects received 800mg TP, 11mg TID NT or 200 mg×2 weeks TC. Lab values were evaluated at baseline and follow-up. The primary outcome was changes in serum 17-OHP. Secondary outcome was changes in serum T. Data analyzed via Kruskal Wallis and Chi square test.Trials: NCT04439799 (June 19,2020) & NCT04523480 (August 21,2020). RESULTS: Median participant age was 45 years old, with overall baseline 17-OHP of 46 and serum testosterone of 223.5 ng/dL. Serum 17-OHP significantly decreased in all three TRT regimens (Table 1). The 4-month change in 17-OHP in the NT group (−33.3% from baseline) was less than the change seen in TC (−65.3% from baseline) or TP (−44% from baseline) (p=0.005). All T formulations increased serum testosterone levels at follow-up, with the largest increase seen in TC (+157.6%), followed by NT (+114.3%), and TP (+79.6%) (p=0.005). CONCLUSIONS: Short-acting nasal testosterone appears to decrease 17-OHP less than long-acting T formulations. All modalities saw significant increases in serum T levels at follow-up. Intranasal T and other short acting T formulations may better preserve ITT and be beneficial for hypogonadal men seeking to maintain fertility potential while on TRT. Source of Funding: Investigator Initiated Grant funded by Acerus Pharmaceuticals and Empower Pharmacy (RR). This work was also supported by National Institutes of Health Grant R01 DK130991 and Clinician Scientist Development Grant from the American Cancer Society to RR © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e580 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Parris Diaz More articles by this author Rohit Reddy More articles by this author Ruben Blachman-Braun More articles by this author Alexandra Dullea More articles by this author Isaac J. Zucker More articles by this author Daniel C. Gonzalez More articles by this author Eliyahu Kresch More articles by this author Ranjith Ramasamy More articles by this author Expand All Advertisement PDF DownloadLoading ...

MP38-19 PREVALENCE OF TESTOSTERONE DEFICIENCY AND SECONDARY POLYCYTHEMIA IN MEN WITH OBSTRUCTIVE SLEEP APNEA

You have accessJournal of UrologyCME1 May 2022MP38-19 PREVALENCE OF TESTOSTERONE DEFICIENCY AND SECONDARY POLYCYTHEMIA IN MEN WITH OBSTRUCTIVE SLEEP APNEA Chase Carto, Sirpi Nackeeran, and Ranjith Ramasamy Chase CartoChase Carto More articles by this author , Sirpi NackeeranSirpi Nackeeran More articles by this author , and Ranjith RamasamyRanjith Ramasamy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002592.19AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Secondary polycythemia is a well-known complication of obstructive sleep apnea (OSA), a disorder that increases in incidence with age. Similarly, increased recognition of low testosterone and its associated symptoms in aging men has led to increased administration of testosterone replacement therapy (TRT), which has also been associated with secondary polycythemia. We hypothesized that given the common association of aging with OSA and testosterone deficiency, a significant percentage of men with OSA would carry concurrent diagnoses of testosterone deficiency and secondary polycythemia. The objective of this study was to investigate the epidemiology of men with OSA, secondary polycythemia, and testosterone deficiency in men using a large electronic health record network. METHODS: We performed a real-time search for adult men (18+) with a diagnosis of obstructive sleep apnea (ICD-10-CM G47.33) in the TriNetX network, which provides health record data for more than 75 million patients from 48 healthcare organizations. This cohort was used to separately identify men with testosterone deficiency, as defined by those with a diagnosis of testicular dysfunction or hypofunction (ICD-10-CM E29, E29.1), and secondary polycythemia (ICD-10-CM D75.1). Lastly, we queried the network for men with all three diagnoses: OSA, testosterone deficiency, and secondary polycythemia. All men with diagnosis of polycythemia vera or familial erythrocytosis were removed from the analysis to account for men with concurrent primary polycythemia. RESULTS: A total of 1,166,640 men with a diagnosis of OSA were identified in the network. Mean age of men with OSA was 61 [46 – 76]. Among men with OSA, 6.0% of men had testosterone deficiency (TT <300ng/dL), and 1.3% of men had associated polycythemia (Hct >52%). Interestingly, only 3,562 (0.3%) of men in the network had all three diagnoses of obstructive sleep apnea, testosterone deficiency, and secondary polycythemia. Strengths of this study include use of a large and heterogenous study population and adjustment for other causes of polycythemia. Limitations include lack of data on chronology of diagnoses, inconsistent data recording, and missing data. CONCLUSIONS: In a large, cross-sectional, multicenter electronic health record study, we found that 6.0% of men with OSA will have testosterone deficiency. Reassuringly, only 0.3% of men carry all diagnoses of obstructive sleep apnea, testosterone deficiency, and secondary polycythemia. Nevertheless, men with OSA who receive TRT warrant close follow-up for potentially dangerous increases in hematocrit when considering treatment for testosterone deficiency. Source of Funding: This work was supported by National Institutes of Health Grant R01 DK130991 and the Clinician Scientist Development Grant from the American Cancer Society to Ranjith Ramasamy © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e625 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Chase Carto More articles by this author Sirpi Nackeeran More articles by this author Ranjith Ramasamy More articles by this author Expand All Advertisement PDF DownloadLoading ...

Delivery of TAPI‐1 via Neutrophil Membrane Fusogenic Liposomal Nanoparticles Can Effectively Ameliorate Cardiac Dysfunction in Heart Failure

Nanomedicine has been widely employed as a site‐specific and target‐oriented drug delivery tactic in treating heart failure (HF). Ischemia‐induced inflammation plays a robust role in the development of HF and neutrophil is one of the main immune cell types that rapidly infiltrate into the inflamed tissues after myocardial infarction (MI). In the present study, we examined the efficacy of a formation of biomimetic nanoparticles that were prepared by fabricated TAPI‐1 loaded liposomes coated with neutrophil membranes in the treatment of HF. Male SD rats underwent coronary artery ligation (CL) to induce HF. These HF rats were treated with intravenous injection of TAPI‐1‐loaded biomimetic nanoparticles (n=6) or TAPI‐1 alone (n=6) for 4 weeks. The left ventricular (LV) function was assessed by echocardiography within 24 hours and 4 weeks after CL. The nanoparticles were evaluated prior to the treatments. Compared with non‐coated formation, neutrophil membrane‐coated nanoparticles have high uptake into LPS‐stimulated cardiac cells or endothelial cells. Moreover, the coated nanoparticles exhibited higher cell‐specific targeting potential and internalization capacity than non‐coated nanoparticles in LPS‐stimulated endothelial cells. Small animal imaging showed that DiR‐labeled nanoparticles were mainly accumulated in the injured heart after MI. Compared with HF rats treated with TAPI‐1 alone, the HF rats treated with TAPI‐1‐loaded nanoparticles had significantly (* p&lt;0.05) improved anatomic indicators with reduced ratio (%) of heart/body weight (0.34 ± 0.042* vs. 0.48± 0.11) and lung/body weight (0.84 ± 0.19* vs. 1.14 ± 0.15). Additionally, one 4‐week treatment with TAPI‐1‐loaded biomimetic nanoparticles reduced LV end systolic volume (0.67±0.032* vs. 0.83±0.036, mL) when compared with the TAPI‐1 alone. While LV stroke volume (LVSV) did not change significantly in IV TAPI‐1‐treated HF animals, LVSV (0.47±0.043* vs. 0.27±0.029, mL) was significantly increased in HF rats treated with TAPI‐1‐loaded nanoparticles at 4 weeks vs. 24 hours after CL. LV end diastolic volume, LV ejection fraction, and LV infarction zone didn’t exhibit significant differences between two treatment groups. Cardiac hemodynamic measurements revealed that the HF rats treated with TAPI‐1‐loaded nanoparticles had decreased LV end diastolic pressure (10.53±1.67* vs. 19.01±3.32, mmHg) and elevated dP/dtmax(8202.65±658.09* vs. 5994.47±630.81) compared with the HF rats‐treated with TAPI‐1 alone. Histological evaluation indicated that the fibrosis areas surrounding the infarction scar were significant reduced (15.97±1.84* vs. 22.86±2.27, %) in HF rats treated with TAPI‐1‐loaded nanoparticles vs. TAPI alone. Flow cytometric analysis also demonstrated that treatment with TAPI‐1‐loaded nanoparticles improved inflammatory conditions indicated by increased ratio of CD4+/CD8+ T Cells (2.40±0.67* vs. 1.56±0.21) in the blood and reduced percentage of leukocytes in the heart of HF. These data indicate that neutrophil membrane fusogenic liposomal nanoparticle is an effective drug delivery approach in treating this devastating disease. (Supported by NIH grants R01 HL‐139521 &amp; HL‐155091 to SGW, S10 OD019941 to RW)

Role of Macrophage accumulation activates NLRP3 inflammasome induced IL-18 pathway in EoE pathogenesis

Abstract Objective Most studies implicated eosinophilic esophagitis (EoE) to the induced esophageal eosinophilia mediated inflammation and fibrosis. However, yet the initial mechanism of allergen-induced eosinophils generation is yet not understood. Our current study provides stepwise pathway operating following the interaction of allergen and antigen presenting cells to induce EoE pathogenesis. Methods We performed immunostaining, RT-PCR measure transcript levels and Masson trichrome staining for collagen accumulation. Results We show that allergen-challenge promotes induced macrophage accumulation and epithelial cells hyperplasia in esophagus. Further, we observed that NLRP3 is induced in both macrophage and epithelial cells. The NLRP3 is the source of IL-18 and we show induced levels of IL-18 in allergen challenge mice that generate tissue eosinophilia in experimental model of EoE. Further, we observed a similar induced NLRP3 in the accumulated macrophage and epithelial cells along with induced IL-18 in the esophageal biopsies of human EoE. Mechanistically, to substantiate the role of macrophages, epithelial cells, and IL-18 role in EoE pathogenesis, we experimentally induced EoE in wild type, GM-CSF−/−, IL-18−/− mice. We show a similar NLRP3 expression with no IL-18 induction in IL-18−/− mice compare to induced NLRP3 and IL-18 in wild-type and significantly reduced NLRP3 and IL-18 in GM-CSF−/− mice. The NLRP3 regulated IL-18 induction or reduction is consistent with the tissue accumulated eosinophila and promoting esophageal remodeling and fibrosis in murine model of EoE. Conclusions The findings show that epithelial cells and macrophage derived NLRP3 regulated IL-18 is critical in promoting EoE pathogenesis. Supported by NIH grant R01 AI080581

Glutamine addiction in NKT cells is regulated by AMPK-mTORC1 axis

Abstract Cellular metabolism is essential in dictating conventional T cell development and function, but its role in natural killer T (NKT) cells has not been well studied. We have previously shown that NKT cells operate distinctly different metabolic programming from CD4 T cells, including a strict requirement for glutamine metabolism to regulate NKT cell homeostasis. However, the mechanisms by which NKT cells regulate glutamine metabolism for their homeostasis and effector functions remain unknown. In this study, we report that steady state NKT cells have higher glutamine levels than CD4 T cells and NKT cells increase glutaminolysis upon activation. Among its many metabolic fates, NKT cells use glutamine to fuel the tricarboxylic acid cycle and glutathione synthesis, and glutamine-derived nitrogen enables protein glycosylation via the hexosamine biosynthesis pathway (HBP). Each of these functions of glutamine metabolism was found to be critical for NKT cell survival and proliferation. Furthermore, we demonstrate that glutaminolysis and the HBP differentially regulate IL-4 and IFNg production. Finally, glutamine metabolism appears to be controlled by AMP-activated protein kinase (AMPK)-mTORC1 signaling. These findings highlight a unique metabolic requirement of NKT cells which can be potentially serve as an effective immunotherapeutic agent against certain nutrient restricted diseases. This work was supported in part by National Institutes of Health Grants R01 AI121156 and R01 AI148289 (to C-H.C.).

Alloprimed antibody-suppressor CD8+ T cells preferentially kill alloprimed germinal center B cells

Abstract Antibody-mediated rejection is a significant contributor to transplant rejection. Our prior work identified a novel subset of CXCR5+CD8+ T cells (CD8+ TAb-supp cells) that suppresses alloantibody production by mediating cytotoxic killing of IgG+ alloantibody-producing B cells. The purpose of these studies was to investigate which B cell subsets are susceptible to CD8+ TAb-supp cell-mediated cytotoxicity. Alloprimed CXCR5+CD8+ T cells were isolated from splenocytes of C57BL/6 mice on day 7 following allo-lysate stimulation. Various target cells (alloprimed B cell subsets, macrophages, and dendritic cells) were isolated from CD8 KO mice on day 7 following allo-lysate stimulation. Alloprimed CD8 KO mice are known to be high alloantibody producers from our prior studies. Target cells were tested in co-cultures to evaluate their susceptibility to CXCR5+CD8+ T cell-mediated in vitro cytotoxicity. Alloprimed CXCR5+CD8+ T cells exhibited significantly higher in vitro cytotoxicity towards alloprimed GC B cells (IgG+GL-7+Fas+; 11.4±1.0%) than towards other antibody producing B cell subsets, including alloprimed extrafollicular plasmablasts (IgG+CD138+; 0.9±0.3%) and plasma cells (IgG−CD138+; 1.6±0.5%; p&amp;lt;0.0001 for both). In addition, CXCR5+CD8+ T cells did not kill alloprimed dendritic cells (MHCII+CD11c+; 1.5±0.7) or macrophages (F4/80+; 1.7±0.4) in co-culture. No significant cytotoxicity was observed in control co-cultures using third party-primed CXCR5+CD8+ T cells (1.1±0.2), naïve CD8+ T cells (1.2±0.3%), or alloprimed CXCR5-negative CD8+ T cells (0.6±0.3%). These data show that alloprimed antibody-suppressor CXCR5+CD8+ T cells preferentially mediate cytotoxic killing of alloprimed GC B cells. This work was supported by a National Institutes of Health R01 grant AI083456 (to GLB), CA016058, UL1TR002733, the OSU Division of Transplant Surgery, and the OSU College of Medicine.

Immunogenetic determinants of HSV-2 infection and disease

Abstract Herpes simplex virus-2 (HSV-2) is one of the most prevalent sexually transmitted infections, and can result in life-long, chronic disease. Disease severity, frequency of reactivation, and shedding rates vary between individuals, though little is known about how host genes regulate tissue-specific immune responses. We have previously used the Collaborative Cross (CC) mouse model system, which incorporates the extent of genetic variation found in the human genome, to better model the diversity of outcomes found in human viral infections, so we next probed the CC to identify host genetic regions that regulate viral shedding and disease following HSV-2 infection, as well as tissue-specific immune responses. We performed a screen of mice from different CC strains to assess viral titers and disease following vaginal HSV-2 infection, and then used this data to perform quantitative trait loci (QTL) mapping to identify chromosomal regions linked to vaginal viral shedding rates and levels, as well as virus-associated clinical disease. In parallel experiments, we assessed lymphoid, nervous system, and mucosal immune cell frequencies at innate, adaptive, and memory response timepoints. We observed a distinctive suppressive signature on vaginal Tregs compared to lymph node Tregs, at various times following HSV-2 infection. Additionally, these suppressive responses varied in mice with either higher viral titers, or more tissue inflammation, highlighting the interplay between host immune response and viral infection kinetics. Understanding host factors that contribute to HSV shedding, clinical disease, and immune responses may provide critical insights for developing new preventive strategies or interventions to HSV-2 infection. Supported by grant R21 AI152559-01 from the NIH

Regulation of myeloid-derived suppressor cell expansion by TRAF3 during chronic inflammation

Abstract Myeloid-derived suppressor cells (MDSCs) expand under pathological conditions and orchestrate the immunosuppressive and inflammatory network, and thus are recognized as a prime therapeutic target of human cancers and inflammatory diseases. In this study, we identified the tumor suppressor TRAF3 as a critical regulator of MDSC expansion. We found that myeloid cell-specific Traf3-deficient (M-Traf3−/−) mice exhibited MDSC hyperexpansion during chronic inflammation. Using mixed bone marrow chimeras, we demonstrated that TRAF3-mediated regulation of MDSC expansion involves cell-extrinsic mechanisms. To delineate the environmental factors that induce the aberrant expansion of MDSCs in M-Traf3−/− mice, we performed cytokine protein array analyses and in vivo neutralization experiments. Our results revealed that G-CSF is a major driver of MDSC hyperexpansion in M-Traf3−/− mice during chronic inflammation. Furthermore, we found that G-CSF was mainly produced by CCR4+ inflammatory monocytes and macrophages in mice during chronic inflammation. Taken together, our findings provide novel insights into the cellular and molecular mechanisms underlying TRAF3-mediated regulation of MDSC expansion, and open up new avenues for the development of MDSC-targeting therapeutics in disease conditions. This study was supported by the National Institutes of Health grant R21 AI128264 (P. Xie), a New Jersey Commission on Cancer Research (NJCCR) grant DCHS19CRF005 (P. Xie), a Pilot Award of Cancer Institute of New Jersey through Grant Number P30CA072720 from the National Cancer Institute (P. Xie), and a Busch Biomedical Grant (P. Xie).

Suppression of local IFN-I by commensal microbiota-derived butyrate impairs antitumor effects of ionizing radiation

Abstract The antitumor effects of ionizing radiation (IR) are mediated in part through activation of innate and adaptive immunity. Here, we report that gut microbiota influences tumor control following IR. Vancomycin decreased the abundance of butyrate-producing gut bacteria, accompanied with reduction of butyric acid in circulation and tumor tissues, and enhanced antitumor responses to IR. Oral administration of Lachnospiraceae, a family of vancomycin-sensitive bacteria, associated with increased systemic and intratumoral butyric acid levels and impaired the efficacy of IR in germ-free (GF) mice. Local butyrate inhibited STING-activated type I IFN expression in dendritic cells (DCs) through blockade of TBK 1 and IRF3 phosphorylation, which abrogated IR-induced tumor-specific cytotoxic T cell immune responses without directly protecting tumor cells from radiation. Our findings demonstrate that the selective targeting of butyrate-producing microbiota may provide a novel therapeutic option to enhance tumor radiation sensitivity. This work was supported by a grant from the Ludwig Foundation to R.R. Weichselbaum, and National Institutes of Health/National Cancer Institute Provocative Questions grants R21 CA227528 and R21 CA231273-01 (R.R. Weichselbaum).