Citations

Abstract Senescent cell accumulation contributes to the progression of age-related disorders including Alzheimer’s disease (AD). Clinical trials evaluating senolytics, drugs that clear senescent cells, are underway, but lack standardized outcome measures. Our team recently published data from the first open-label trial to evaluate senolytics (dasatinib plus quercetin) in AD. After 12-weeks of intermittent treatment, we reported brain exposure to dasatinib, favorable safety and tolerability, and modest post-treatment changes in cerebrospinal fluid (CSF) inflammatory and AD biomarkers using commercially available assays. Herein, we present more comprehensive exploratory analyses of senolytic associated changes in AD relevant proteins, metabolites, lipids, and transcripts measured across blood, CSF, and urine. These analyses included mass spectrometry for precise quantification of amyloid beta (Aß) and tau in CSF; immunoassays to assess senescence associated secretory factors in plasma, CSF, and urine; mass spectrometry analysis of urinary metabolites and lipids in blood and CSF; and transcriptomic analyses relevant to chronic stress measured in peripheral blood cells. Levels of Aß and tau species remained stable. Targeted cytokine and chemokine analyses revealed treatment-associated increases in inflammatory plasma fractalkine and MMP-7 and CSF IL-6. Urinary metabolites remained unchanged. Modest treatment-associated lipid profile changes suggestive of decreased inflammation were observed both peripherally and centrally. Blood transcriptomic analysis indicated downregulation of inflammatory genes including FOS, FOSB, IL1β, IL8, JUN, JUNB, PTGS2. These data provide a foundation for developing standardized outcome measures across senolytic studies and indicate distinct biofluid-specific signatures that will require validation in future studies. ClinicalTrials.gov: NCT04063124.

Abstract Migraine and sleep disorders are common co-morbidities. Patients frequently link their sleep to migraine attacks suggesting a potential causal relationship between these conditions. However, whether migraine pain promotes or disrupts sleep or whether sleep disruption can increase the risk of migraine remains unknown. We assessed the potential impact of periorbital allodynia, a measure consistent with migraine-like pain, from multiple preclinical models on sleep quantity and quality. Additionally, we evaluated the possible consequences of sleep deprivation in promoting susceptibility to migraine-like pain. Following the implantation of electroencephalogram/electromyography electrodes to record sleep, mice were treated with either single or repeated systemic injections of nitroglycerin at the onset of their active phase (i.e. nocturnal awake period). Neither single nor repeated nitroglycerin affected the total sleep time, non-rapid eye movement sleep, rapid eye movement sleep, sleep depth or other measures of sleep architecture. To account for the possible disruptive effects of the surgical implantation of electroencephalogram/electromyography electrodes, we used immobility recordings as a non-invasive method for assessing sleep-wake behaviour. Neither single nor repeated nitroglycerin administration during either the mouse sleep (i.e. daylight) or active (i.e. night) periods influenced immobility-defined sleep time. Administration of an inflammatory mediator mixture onto the dura mater at either sleep or active phases also did not affect immobility-defined sleep time. Additionally, inhalational umbellulone-induced migraine-like pain in restraint-stressed primed mice did not alter immobility-defined sleep time. The possible influence of sleep disruption on susceptibility to migraine-like pain was evaluated by depriving female mice of sleep over 6 h with novel objects, a method that does not increase circulating stress hormones. Migraine-like pain was not observed following acute sleep deprivation. However, in sleep-deprived mice, subthreshold doses of systemic nitroglycerin or dural calcitonin gene-related peptide induced periorbital cutaneous allodynia consistent with migraine-like pain. Our data reveal that while migraine-like pain does not significantly disrupt sleep, sleep disruption increases vulnerability to migraine-like pain suggesting that a therapeutic strategy focused on improving sleep may diminish migraine attacks.

Abstract Introduction This study investigated the efficacy and safety of neoadjuvant chemotherapy for locally advance penile squamous cell carcinoma for which current evidence is lacking. Methods Included patients had locally advanced penile squamous cell carcinoma with clinical lymph node metastasis treated with at least 1 dose of neoadjuvant chemotherapy prior to planned consolidative lymphadenectomy. Objective response rates were assessed using Response Evaluation Criteria in Solid Tumors v1.1. The primary and secondary outcomes were overall survival and progression-free survival, estimated by the Kaplan–Meier method. Treatment-related adverse events were graded per the Common Terminology Criteria for Adverse Events v5.0. Results A total of 209 patients received neoadjuvant chemotherapy for locally advanced and clinically node-positive penile squamous cell carcinoma. The study population consisted of 7% of patients with stage II disease, 48% with stage III, and 45% with stage IV. Grade 2 treatment-related adverse events occurred in 35 (17%) patients, and no treatment-related mortality was observed. Of the patients, 201 (97%) completed planned consolidative lymphadenectomy. During follow-up, 106 (52.7%) patients expired, with a median overall survival of 37.0 months (95% confidence interval [CI] = 23.8 to 50.1 months) and median progression-free survival of 26.0 months (95% CI = 11.7 to 40.2 months). Objective response rate was 57.2%, with 87 (43.2%) having partial response and 28 (13.9%) having a complete response. Patients with objective response to neoadjuvant chemotherapy had a longer median overall survival (73.0 vs 17.0 months, P < .01) compared with those who did not. The lymph node pathologic complete response rate was 24.8% in the cohort. Conclusion Neoadjuvant chemotherapy with lymphadenectomy for locally advanced penile squamous cell carcinoma is well tolerated and active to reduce the disease burden and improve long-term survival outcomes.

Abstract Introduction Since its description in 1743 by François de la Peyronie, penile fibromatosis continues to present an elusive pathogenesis. One of its distinguishing characteristics is the excessive proliferation of fibrogenic myofibroblasts, resulting in the formation of dense fibrous plaques that disrupt the normal cytoarchitecture of the tunica albuginea, giving rise to penile abnormalities. The possible clonal nature of the disease has been investigated by many studies and some have identified occasional karyotypic abnormalities, most commonly involving chromosomes 7, 8, 17, 18, X and Y (Mulhal JP et al 2004). Objective This study aimed to comprehensively analyze the genomic landscape of Peyronie's disease in a significant number of cases using a whole genome copy number variation assay (CNV). The objective was to identify any recurrent chromosomal gains or losses that could shed light on the pathogenesis of this disease, including the potential identification of an underlying clonal process. Methods Thirty-nine surgical specimens were retrospectively examined from men diagnosed with Peyronie's disease, collected between 1992 and 2022 (median age: 56 years, range: 17-73). The study was conducted under the approval of the Institutional Review Board (IRB # 22-006924). Tissue samples were fixed in 10% buffered formalin, embedded in paraffin, and subsequently subjected to histologic review and DNA analysis. DNA extraction followed standard protocols. Among the samples, 22 contained high-quality material suitable for CNV array analysis using the OncoScan SNV array, which encompassed approximately 900 genes/loci with a resolution of 50-125 kb. Additionally, the array included a somatic mutation panel targeting nine cancer-related genes. Results No significant loss or gain of chromosomal material was identified in the twenty-two cases analyzed. However, the somatic mutation panel revealed potential unconfirmed DNA mutations in NRAS, EGFR, and TP53. Ongoing studies are being conducted to confirm these mutations. Conclusions The etiology and pathogenesis of Peyronie's disease remain unresolved, presenting a challenging area of research. However, the findings of this study provide evidence that major recurrent losses or gains of chromosomal material are unlikely to play a significant role in the pathogenesis of this disease. While shedding light on this aspect, it is crucial to acknowledge that further research is necessary to confirm and expand upon these initial observations. Disclosure No.

Subharmonic aided pressure estimation (SHAPE) is a technique that utilizes subharmonic signals from microbubble contrast agents for pressure estimation. Validation of the SHAPE technique relies on synchronous measurements of in vivo pressures using contrast microbubbles and a pressure catheter (reference standard). For the guidance and placement of pressure catheter in vivo, iodinated contrast is used with fluoroscopy. Therefore, during data acquisition for validation studies of the SHAPE technique, both contrast microbubbles and iodinated contrast are present simultaneously within the vasculature. This study aims to elucidate the effects of iodinated contrast (Visipaque, GE HealthCare) on subharmonic signal amplitude from contrast microbubbles (Definity, Lantheus Medical Imaging, Inc.). In an acrylic water tank, 0.06 mL of Definity and varied amounts of Visipaque (0.14, 0.43, 0.85, and 1.70 mL) were added to 425 mL of deionized water. Ultrasound scanning was performed with a SonixTablet scanner (BK Medical Systems) using optimized parameters for SHAPE with Definity ( f transmit/receive = 3.0/1.5 MHz; chirp down pulse). Subharmonic data was acquired and analyzed at 9 different incident acoustic outputs ( n = 3). Results showed an increase in subharmonic signal amplitude from Definity microbubbles in the presence of 0.14 mL Visipaque by 2.8 ± 1.3 dB ( p < .001), no change with 0.85 mL Visipaque (0.7 ± 1.2 dB; p = .09) and a decrease in subharmonic amplitude in the presence of 1.70 mL Visipaque by 1.9 ± 0.7 dB ( p < .001). While statistically significant effect on subharmonic signal amplitude of Definity microbubbles was noted due to the mixture, the magnitude of the effect was minimal (~2.8 dB) and unlikely to impact in vivo SHAPE measurements.

The LMNA gene encodes lamin A and lamin C, which play important roles in nuclear organization. Pathogenic variants in LMNA cause laminopathies, a group of disorders with diverse phenotypes. There are two main groups of disease-causing variants: missense variants affecting dimerization and intermolecular interactions, and heterozygous substitutions activating cryptic splice sites. These variants lead to different disorders, such as dilated cardiomyopathy and Hutchinson–Gilford progeria (HGP). Among these, the phenotypic terms for LMNA-associated cardiocutaneous progeria syndrome (LCPS), which does not alter lamin A processing and has an older age of onset, have been described. Here, we present the workup of an LMNA variant of uncertain significance, NM_170707.2 c. 4G>A, p.(Glu2Lys), in a 36-year-old female with severe calcific aortic stenosis, a calcified mitral valve, premature aging, and a family history of similar symptoms. Due to the uncertainty of in silico predictions for this variant, an assessment of nuclear morphology was performed using the immunocytochemistry of stable cell lines to indicate whether the p.(Glu2Lys) had a similar pathogenic mechanism as a previously described pathogenic variant associated with LCPS, p.Asp300Gly. Indirect immunofluorescence analysis of nuclei from stable cell lines showed abnormal morphology, including lobulation and occasional ringed nuclei. Relative to the controls, p.Glu2Lys and p.Asp300Gly nuclei had significantly (p < 0.001) smaller average nuclear areas than controls (mean = 0.10 units, SD = 0.06 for p.Glu2Lys; and mean = 0.09 units, SD = 0.05 for p.Asp300Gly versus mean = 0.12, SD = 0.05 for WT). After functional studies and segregation studies, this variant was upgraded to likely pathogenic. In summary, our findings suggest that p.Glu2Lys impacts nuclear morphology in a manner comparable to what was observed in p.Asp300Gly cells, indicating that the variant is the likely cause of the LCPS segregating within this family.

Purpose: To compare left-ventricular and aortic pressures obtained using fluid-filled and high-fidelity solid-state pressure catheters in subjects undergoing left heart catheterization. Methods: Twenty subjects scheduled for a left heart catheterization were enrolled and 18 subjects completed this IRB-approved study. During catheterization, left-ventricular (LV) and aortic (AO) pressures were obtained using a fluid-filled pressure catheter (standard-of-care) and a high-fidelity solid-state pressure catheter synchronously. The pressure tracings were analyzed by two independent readers to measure LV systolic (LVSP), LV minimum diastolic (LVMDP), LV end diastolic (LVEDP), AO systolic (AOSP), and AO diastolic (AODP) pressures. Derivatives of the pressure waveforms were computed to estimate the isovolumic contraction and relaxation rates (peak ± dp/dt). The measurements were made over multiple cardiac cycles and averaged results were used for comparisons using repeated measures analysis of variance, post-hoc tests with Bonferroni corrections and Bland-Altman plots. Results: A significant main effect of the pressure catheter was noted for LVSP, LVMDP and AOSP (p < 0.025). The LVSP and AOSP measured with fluid-filled pressure catheters were higher by 6.6 ± 6.9 mmHg and 4.6 ± 5.2 mmHg in comparison to the high-fidelity solid-state pressure catheter assessments. In contrast, the LVMDP measurements were 3.5 ± 5.7 mmHg lower than the solid-state pressure catheter measurements. When evaluating the differences between readers, the maximum difference was noted for LVEDP measurements (3.1 ± 2.4 mmHg; p ≤ 0.001). The differences in isovolumic contraction rate (66.4 ± 116.0 mmHg/s) and relaxation rate (60.5 ± 113.5 mmHg/s) were not significantly different between the two catheter systems after Bonferroni corrections for multiple comparisons (p > 0.06). Bland-Altman analysis revealed a bias ranging from 0.8 to 6.6 mmHg for all the pressure measurements. Conclusions: There are differences in LVSP, LVMDP, and AOSP measured using fluid-filled and high-fidelity solid-state pressure catheters. Clinicians should be aware that the fluid-filled catheters commonly overestimate the true systolic pressures in the left ventricle and aorta.