Louis Stokes Cleveland VA Medical Center

Seventy-five years of research funding in benign Urology from the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) at the National Institutes of Health has greatly advanced not only our knowledge of etiology and mechanisms of nononcologic urological conditions but has also advanced new technologies and treatments to improve clinical care. To illustrate the profound impact of NIDDK funding on research and clinical care, we undertook a survey of grants funded by NIDDK in the NIH Reporter website with a focus on translation of lower urinary tract health and disease research to the clinic. We highlight three examples of technologies whose development was funded by NIDDK: advanced artificial urinary sphincters, urodynamics, and neuromodulation. These examples demonstrate the difficulties in bringing innovations to clinical care, as well as successes. In addition, they illustrate how public investment through NIDDK funding can improve treatment outcomes, diagnostic capabilities, and provide multiple therapeutic options.

Spinal cord injury typically leads to neurogenic bladder dysfunction, often including detrusor-sphincter dyssynergia. Sacral anterior root stimulation can empty the neurogenic bladder, but this emptying can be impeded by reflex contractions of the urethral sphincter. The sacral sensory roots are typically transected (rhizotomy) to reduce these reflex contractions, but this rhizotomy also impairs desirable reflexes (e.g., sexual function) and sacral sensation, if present. Preclinical experiments have shown that sacral nerve stimulation at 600 Hz can promote bladder emptying while reducing urethral sphincter activity. In this proof-of-concept study, we tested 600-Hz sacral nerve stimulation to limit urethral sphincter activity in individuals with spinal cord injury who were already implanted with a sacral anterior root stimulation device. Bladder pressure, urethral pressure, rectal pressure, anal pressure, and pelvic floor electromyogram were measured in response to stimulation at 20 or 600 Hz. Stimulation was feasible and well-tolerated, and frequency appeared to have an effect on lower urinary tract and bowel function. In one participant, stimulation at 600 Hz produced a significant decrease in urethral pressure compared to 20-Hz stimulation. Further work is needed to explore the potential for this approach to improve bladder emptying efficiency for individuals with spinal cord injury and detrusor-sphincter dyssynergia.

This study focuses on developing an efficient large-area graphene transfer method that combines high-throughput and precise laser engraving, simple polymer casting, and water immersion to fabricate conductive graphene and biodegradable polymer-based implantable flexible electronic devices. The low-temperature treatment of graphene sheets on a glass substrate reduced graphene sheet roughness and increased hydrophobicity, enabling facile and high-efficiency (∼100%) large-area graphene transfer to a flexible polymer substrate. This method also benefited from differences in the work of adhesion at the graphene sheet/glass substrate and the graphene sheet/flexible polymer substrate interfaces. The transferred graphene sheets showed stability, structural integrity, and high conductivity (∼40 Ω/sq sheet resistance) under in vitro and in vivo mimicking conditions. The low-temperature-treated and laser-engraved conductive graphene patterns, transferred on a flexible and biodegradable polymer substrate, demonstrated in vitro cytocompatibility on different cells. Two flexible electronic devices (1─a graphene coil-integrated electrode cuff and 2─an interdigitated graphene cuff-integrated piezoelectric device) were fabricated using the developed method, and both demonstrated functionality and proof of concept by generating output voltages that can enhance cell/tissue regeneration. In addition, the ease of handling, ex vivo implantation, and feasibility of suturing were demonstrated by performing implantation surgeries on the pudendal nerve in cadaveric rats. Overall, this promising large-area graphene transfer method can be used to fabricate biodegradable, implantable devices that can serve as interfaces to stimulate cells and tissues for regeneration and repair.

Repurposing existing medicines to target disease-associated genes represents a promising strategy for developing effective treatments for complex diseases. However, progress has been hindered by a lack of viable candidate drug targets identified through genome-wide association studies. Gene-based association tests provide a more powerful alternative to traditional SNP-based methods, yet current approaches often fail to leverage shared heritability across populations and to effectively integrate functional genomic data. To address these challenges, we develop GenT and its various extensions, comprising a framework of gene-based tests utilizing summary-level data from genome-wide association studies. Using GenT, we identify 16, 15, 35, and 83 candidate genes linked to Alzheimer's disease, amyotrophic lateral sclerosis, major depression, and schizophrenia, respectively, not detected by Genome-Wide Association Studies (GWAS). Additionally, we use our multi-ancestry gene-based test (MuGenT) to identify 28 candidate genes associated with type 2 diabetes. By integrating brain expression and protein quantitative trait loci into our analysis, we identify 43 candidate genes associated with Alzheimer's disease that have supporting xQTL evidence. We also perform experimental assays to demonstrate that the NTRK1 inhibitor GW441756 significantly reduces tau hyper-phosphorylation (including p-tau181 and p-tau217) in Alzheimer's disease patient-derived iPSC neurons, providing mechanistic support for our predictions.

Mutations in TULP1 are associated with early-onset forms of inherited retinal degenerations (IRDs). Evidence from Tulp1-/- mice indicates that TULP1 plays a role in photoreceptor protein trafficking. Here we generated two novel knock-in mouse models, each expressing the ortholog to a human IRD-causing homozygous missense TULP1 mutation to: 1) better recapitulate IRD patients' gene dosage and spatiotemporal degeneration, 2) determine the pathological disease mechanism, and 3) evaluate mutations affecting different domains of the protein. The Tulp1F492L model carries a mutation affecting a conserved amino acid in the C-terminal tubby domain, whereas the Tulp1D89Y model carries the only homozygous mutation located outside the tubby domain. In both mutant retinas, TULP1F492L and TULP1D89Y protein levels and distribution were comparable to WT. Surprisingly, variable retinal phenotypes were observed in the two mutant lines. The Tulp1F492L model displayed rapid photoreceptor degeneration, rod and cone opsin mistrafficking, and abnormal shaped ribbon synapses, similar to Tulp1-/- mice. In contrast, these abnormalities were not seen in the Tulp1D89Y model; indeed, retinal morphology and function was preserved up to 12months, although we noted less RPE pigmentation and dilated structures in the outer plexiform layer at this timepoint. Moreover, building on our prior in vitro results, we observed activation of the IRE1 branch of the endoplasmic reticulum (ER)-unfolded protein response (UPR) complex in Tulp1-/- and Tulp1F492L retinas, identifying ER stress as a key disease mechanism leading to photoreceptor death and as a potential therapeutic target in TULP1-associated forms of IRD.

Cefazolin is the preferred prophylactic antibiotic for many surgical procedures but is often unnecessarily avoided in patients with reported penicillin and/or cephalosporin allergies. This quality improvement project aimed to increase cefazolin use for preprocedural prophylaxis in these patients. Quasi-experimental quality improvement study Safety net healthcare system in the United States Patients with reported penicillin and/or cephalosporin allergies undergoing procedures requiring antibiotic prophylaxis and anesthesiology services from January 2022 to October 2023 Interventions included the development of allergy management algorithms, educational sessions, electronic medical record updates, and audit and feedback by pharmacists The primary outcomes were changes in proportions of procedures for which cefazolin, clindamycin, and vancomycin were used for preprocedural antibiotic prophylaxis. The secondary outcomes were changes in proportions of procedures in which appropriate dose and timing of preprocedural antibiotic prophylaxis were administered. Data from 3414 patients undergoing various procedures requiring antibiotic prophylaxis and anesthesiology services during the study period were analyzed. The use of cefazolin increased from 30% pre-intervention to 73% post-intervention (p < 0.001), while clindamycin use decreased from 51% to 13% (p < 0.001), and vancomycin use decreased from 5.8% to 3.9% (p = 0.012). Appropriate antibiotic dosing increased from 89% to 93% (p = 0.048), and appropriate timing of preprocedural antibiotics improved from 49% to 74% (p < 0.001). A multi-pronged intervention significantly increased cefazolin use in patients with reported penicillin and/or cephalosporin allergies, leading to optimized antibiotic prophylaxis and reduced use of less effective alternatives.

Studies have demonstrated that repeated mRNA vaccination enhances the breadth of neutralization against diverse severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants. However, the development of antibodies capable of neutralizing across the Coronavirinae subfamily is poorly understood. In this study, we analyze serum samples to determine their neutralization breadth and potency and identify their antigenic targets. Using a cohort of older individuals and healthcare workers, we track correlates of broad neutralizing responses, including fusion peptide (FP) antibody elicitation. We find that although broadly neutralizing responses are often a result of receptor-binding domain (RBD)-specific antibodies, a rare subset of donors produce FP-specific broadly neutralizing responses. Interestingly, FP-specific antibodies are not observed in COVID-naive individuals, irrespective of vaccination regimen; rather, they occur following natural infection or vaccine breakthrough. This study highlights which epitope targets underpin broadly neutralizing antibody responses to coronaviruses and suggests that existing vaccines are insufficient to promote the elicitation of FP-directed broadly neutralizing coronavirus antibodies.