University Of Nebraska Medical Center

NSF Awards · FY 2025 · 2025-08

Embedded 3D bioprinting has been extensively utilized to fabricate human organ and tissue analogs; however, creating human tissue constructs with large scale and high cell viability remains a significant challenge. The research goal of this EArly-concept Grant for Exploratory Research (EAGER) project is to establish a new embedded section-by-section bioprinting process aimed at achieving a high cell viability during the fabrication of large-scale human tissue constructs. In this new process, a human-relevant sized construct will be segmented into multiple smaller sections and sequentially printed within a support bath using a short nozzle. Between sections, a special material will be added to the support bath, and its viscosity and flow behavior will be adjusted by changing the temperature. The expected outcomes of this project are 1) identifying cell damage mechanisms in this 3D bioprinting process and 2) elucidating cell damage during the addition of support bath materials. This new 3D bioprinting strategy will facilitate the reconstruction of full-scale human organs and tissues for diverse biomedical applications. The education goal is to enhance the biomanufacturing training through outreach activities, such as bioprinting demonstrations to K-12 students, bioprinting-related curricula for undergraduates and graduates, biomanufacturing-themed activities to attract talented students. This project aims to advance and surpass the state-of-the-art embedded 3D bioprinting processes by ensuring a high initial cell viability of over 80 percent when producing human tissue constructs with dimensions exceeding 20 mm along the printing orientation. Three integrated research objectives will be pursued, including 1) establish a theoretical platform through mechanics modeling and experimentation to identify the cell damage mechanisms at the cellular level and during bioextrusion; 2) elucidate thermal damage to living cells via numerical simulation, modeling, and experiments to optimize key conditions when adding support bath materials; and 3) evaluate the capability of the proposed embedded section-by-section bioprinting process to print large-scale cellular human tissues. The knowledge gained from this project will jump-start the development of advanced support bath materials and innovative 3D bioprinting processes for tissue engineering in the future. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2025-08

Abstract The Lyme Disease pathogen Borrelia burgdorferi is estimated to cause nearly half a million human infections in the U.S. every year. Transmitted by the blacklegged deer tick (Ixodes scpaularis), B. burgdorferi is passed between rodents, ticks and large mammals to complete its complex enzootic cycle. Humans get infected as an incidental host, usually in the springtime by nymph-stage ticks. In this proposal we adapt an established lab-generated in vitro organotypic stratified keratinocyte skin model to induce attachment and feeding of ticks. In Aim 1, we continue to modify the skin to increase tick feeding efficiency and demonstrate transmission and acquisition of B. burgdorferi by nymph-stage ticks. In Aim 2, we characterize the kinetics of bacterial growth and spread within organotypic skin following direct injection and validate that B. burgdorferi responds to environmental cues of the bloodmeal (pH, temperature) to increase expression of mammalian stage virulence factors. This novel organotypic human skin model is highly adaptable and can accommodate introduction of other cell types. For instance, in Aim 1 we introduce endothelial cells, macrophages and extracellular matrix factors into the skin to induce vascularization and provide a biologically-relevant blood delivery system. This is the first demonstration of arthropod feeding on in vitro organotypic human skin, and its development will enable new avenues of research not only for Lyme Disease research, but also for other tick-borne diseases. We expect this model will reduce the need for animals in Lyme Disease research and will provide a system to answer long-standing questions in tick-pathogen biology, such as the role of resident immune cells in containing infection and the contribution of specific bacterial virulence factors to skin colonization and dissemination into the bloodstream. Given the scalable and reproducible nature of this model, we can use it as a platform to screen therapies that could prevent pathogen colonization following a tick bite or prophylactics that would prevent tick feeding.

NSF Awards · FY 2025 · 2025-08

Breast cancer is the second most common malignancy and the second leading cause of cancer death among women in the United States. Previous studies indicated that Black women have disproportionately higher mortality rates in breast cancer in the United States. With artificial intelligence (AI) and machine learning (ML) being increasingly applied to cancer research and clinical decision making, especially for breast cancer detection, diagnosis, prognosis and treatment, cancer data variability would lower the quality and utility of AI/ML models. This project aims to develop a novel AI framework to reduce health outcome variability for breast cancer based on multi-modal genomics data. In the long term, this project will enhance the quality of health outcomes in breast cancer detection, diagnosis, prognosis, and treatment. Previous studies suggested that transfer learning could enhance health outcomes in breast cancer. However, its performance may be reduced when the number of samples is small, or only single-omics data are used. This project will develop novel machine learning and statistical approaches to establish an integrated multi-modal transfer learning framework for breast cancer. The technical aims of this project are divided into three parts. The first aims to establish a multi-omics integration model to enhance breast cancer outcomes. Compared to single-omics data, multi-omics data provide a broader and more complete set of training data for ML models, which can increase the generalizability of the ML model and thus improve the model’s capabilities. The second thrust aims to develop a novel transfer learning framework. Existing transfer learning models require substantial amounts of data for fine-tuning, which are difficult to obtain in clinical settings. To address this concern, this project will develop a multi-modal transformer for pre-training and a new data-augmentation method for the transfer learning framework. The third thrust aims to identify and validate informative biomarkers for breast cancer diagnosis and prognosis. Conventional methods for biomarker identification for breast cancer ignore cancer variability. Developing and utilizing such biomarkers can contribute to more equitable and personalized cancer care. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2025-07

Staphylococcus aureus (S. aureus) is a leading cause of biofilm-associated prosthetic joint infection (PJI) typified by antibiotic tolerance and evasion of immune-mediated clearance. Our laboratory has extensively utilized a model of S. aureus PJI in inbred C57BL/6J mice to define immune attributes that promote biofilm persistence. This includes an abundance of anti-inflammatory granulocytic myeloid-derived suppressor cells (G-MDSCs) that inhibit monocyte/macrophage proinflammatory activity and neutrophil (PMN) killing of S. aureus through robust IL-10 production. Despite these advances, genetic diversity is not captured with inbred mouse strains, and it is well appreciated that genetic variability in humans contributes to heterogeneity in immunity and disease susceptibility. This is reflected in our studies of patients with PJI, where transcriptional profiles of leukocytes recovered from infected tissues displayed significant heterogeneity, even when infected with the same bacterial species. Furthermore, there are wide variations in the numbers of G-MDSCs associated with human PJI tissues. Therefore, despite their utility, inbred mice are not capable of capturing the diverse array of genetic factors that impact disease outcome during human PJI. To address this issue, this proposal leverages the Collaborative Cross (CC), a genetically diverse group of mouse strains representing > 30 million SNPs that were generated by crossing 5 classical inbred lines with 3 wild-derived inbred strains. The genomes of CC mice are fully sequenced, allowing genes of interest to be identified by quantitative trait locus (QTL) mapping to assign genotype-phenotype relationships. Only one report to date has utilized CC strains to examine host genes that influence S. aureus pathogenesis in a sepsis model, which is highly divergent from biofilm pathology. Therefore, due to the lack of comprehensive GWAS studies or the use of CC strains in PJI, little is known about host genes that regulate biofilm growth. We will leverage CC lines in a two-pronged approach to identify the effect of disease- associated QTLs on PJI outcome and the intrinsic genetic determinants that shape PMN acquisition of anti- inflammatory G-MDSC-like characteristics. This is expected to uncover novel host genetic factors that impact PJI pathogenesis, advancing the applicability of our findings to the clinical realm. This proposal will test the hypothesis that genetic diversity contributes to the regulation of biofilm growth and inflammation, which will be examined in the following Specific Aims: 1) Explore the genetic determinants that influence host immunity during PJI using CC mice; and 2) Identify host genetic loci that are critical for PMN acquisition of G-MDSC-like properties in response to S. aureus biofilm. These studies are expected to inform our long-term goal of targeting key host regulatory genes in combination with antibiotics to clear PJI without the need for additional surgeries, reducing the morbidity for patients suffering from these debilitating infections.

NIH Research Projects · FY 2025 · 2025-07

Scientific Abstract By expressing fluorescent reporters, such as green fluorescent protein (GFP) or red fluorescent protein (RFP) downstream of specific promoters or fusing them with endogenous genes, a range of cellular processes can be easily tracked and studied using fluorescence microscopy. Fluorescent reporters become even more powerful when they are tagged with a protein domain that transforms their function into an indicator of specific cellular processes, and they are typically referred to as ‘genetically encoded fluorescent indicators.’ Such indicators have been specifically designed by researchers to detect changes in ion concentration, membrane potential, and pH and have been used to generate transgenic mouse models for in vivo studies. The vast majority of these transgenic mice expressing genetically encoded fluorescent indicators utilize GFP as the fluorescent moiety. Unfortunately, major limitations of GFP-based indicators are phototoxicity and tissue autofluorescence which make it difficult to differentiate signal from the noise. Although RFP-based indicators can overcome these issues and they provide greater tissue penetration depth, there are not many animal-model resources currently available in the red emission range. The goal of this proposal is to develop mouse models expressing RFP-based indicators, characterize them, and make them available to the scientific community. We will utilize a novel clustered regularly interspaced short palindromic repeats (CRISPR)-based genome engineering approach called RE-CREATING (RE-engineering, with CRISPR, of previously Engineered Alleles To Insert New Genes), to generate two conditional knock-in mouse models. A popular and well-studied mouse strain that expresses a green genetically encoded fluorescent indicator will be used as the parental strain. The transgene locus in this parental strain will be re-engineered by swapping the GFP-based indicator for a RFP-based indicator, without disrupting any of the regulatory elements. The newly developed RFP-based indicator mouse models will be suitable to study any cell type of interest simply by crossing them with a Cre driver mouse of choice. Our models will enable imaging of dynamic intracellular processes such as changes in calcium activity and membrane potential, which can be challenging and/or inconsistent across tissues in currently available GFP-based models (due to phototoxicity and autofluorescence). We will use vascular physiology techniques (e.g., pressure myography and sharp microelectrode measurements of membrane potential) to characterize and validate the RFP-based indicator mouse models. Overall, this project will provide a much-needed resource to the scientific community and opens the door for imaging of dual indicator mice and/or allows for imaging of processes not traditionally possible with GFP-based mouse models.

NIH Research Projects · FY 2025 · 2025-07

Project Summary Background: Children born to mothers with human immunodeficiency virus type-1 (HIV-1) infection are on the rise. With an increased coverage of antiretrovirals (ARVs) for pregnant women, especially in in resource-limited countries (RLCs), over one million ARV-exposed-HIV-1-uninfected (HEU) children are born every year. Even though ARVs have helped in preventing fetal HIV-1 infection and infection-associated maternal or fetal mortalities, risks of adverse events in fetuses linked to exposure to ARVs remain a major concern. Importantly effect of gestational ARVs exposure on pre- and post-natal neurodevelopment remain incompletely understood. These include physiological, metabolic, or functional harms to developmental brain of children who were born without chronic structural malformations. This is underscored by the reports of risk of postnatal neurodevelopmental deficits in HIV-1-exposed uninfected (HEU) children associated with, but not limited to, efavirenz (EFV), atazanavir (ATV), didanosine (ddI), or dolutegravir (DTG). Thus, with over a million HEU children born each year, and continuous efforts to develop new potent ARVs, development of a non-invasive bioimaging tool which can help for early-recognition (during gestation) of ARV-linked adverse neurodevelopmental outcomes is timely. Such bioimaging tool upon successful development will aid to define which ARVs have adverse effect on neurodevelopment, uncover altered molecular biomarkers as underlying mechanisms, irrespective of classification of ARVs or of administration through daily pills, long acting injectables or implants. Objective: (1) Implement novel chemical exchange saturation transfer (CEST) magnetic resonance imaging (MRI) to non- invasively track ARVs-linked fetal neurodevelopmental adverse events during gestation in a rodent model. (2) Develop novel 3D variableRFCEST sequence which will acquire data from entire brain with varying RF saturation power that enables the simultaneous acquisition of various metabolites, mobile proteins or peptides in short duration. (3) Uncover altered fetal brain metabolites as an early-stage biomarkers for ARVs-associated developmental neurotoxicity. Our preliminary data: In preliminary work, sensitivity of CEST MRI to detect developmental neuronal impairment in mice embryo brain following ARV-exposure was determined. Hypothesis: We posit that CEST MRI can detect ART-linked fetal neuropathological biomarkers during pregnancy. Research Strategy: (a) Implement CEST MRI to determine the effect of EFV or DTG-based three drug regimens on neuronal membrane lipids and glutamate in developmental pre-frontal cortex in a rodent model. (b) Achieve rigor for CEST MRI sensitivity and specificity for molecular biomarkers. (c) Perform cross-validation of CEST MRI data with pharmacokinetics, metabolomics, and immunohistopathology data. Outcome: Successful implementation of CEST MRI to study fetal neurodevelopment will provide a non-invasive tool with sensitivity and specificity that has a potential to elucidate the underlying mechanisms of ART-induced developmental neurotoxicity.

NSF Awards · FY 2025 · 2025-05

This I-Corps project focuses on the development of an automated system that provides healthcare professionals with up-to-date information about which antibiotics are effective against specific bacteria. Antimicrobial resistance is a major public health problem worldwide, in part due to misuse and overuse of antibiotics. Current methods for tracking bacterial resistance patterns involve static reports that are updated infrequently, sometimes only once per year, leading to outdated guidance for clinicians. This technology allows healthcare providers across various settings including hospitals, clinics, and long-term care facilities to access continuously updated resistance data. The solution helps providers select the most appropriate antibiotics for patient treatment, potentially reducing mortality rates, shortening hospital stays, and lowering healthcare costs. By improving antibiotic selection, the technology contributes to national health objectives related to combating antibiotic resistance. This I-Corps project utilizes experiential learning coupled with a first-hand investigation of the industry ecosystem to assess the translation potential of the technology. This solution is based on the development of a dynamic susceptibility reporting system that transforms laboratory microbiology data into actionable clinical intelligence. The system integrates with existing laboratory information systems and leverages modern web development frameworks to process microbiological data in real-time. Unlike traditional static reports, the technology allows for customized filtering by variables such as patient location, specimen type, and time period, creating tailored guidance for specific clinical scenarios. The system architecture accommodates varying institutional data structures and reporting requirements, enhancing its potential for widespread adoption. Technical innovations include automated data processing that eliminates the labor-intensive manual compilation required for traditional reports and real-time analysis capabilities that reflect current resistance trends rather than historical patterns. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-05

Project Summary/Abstract The broad, long-term objectives of this proposal are to enhance the utility of cellulose-based biomaterials for tissue repair by developing and evaluating a new and innovative composite that address current limitations. Bacterial cellulose hydrogels and extracellular matrices have shown excellent regeneration capabilities in multiple tissue types. However, these materials lack mechanical strength and degradation features needed for specific applications such as bone repair, and have limited options for storage, handling, and sterilization. Plant- derived cellulose in its derivative cellulose acetate (CA) form is capable of creating mechanically competent porous scaffolds that are effective in bone regeneration. However, premade CA scaffolds with defined sizes, shapes, and pore properties present challenges in adapting to complex bone defects. Additionally, the relatively slow degradation rate of cellulose/CA can limit its ability to control factor release and heal bone. Combining CA with CA phthalate (CAP) and nanoclay (NC) has the potential to address some of these weaknesses. This cellulose-based composite forms a putty that can be molded into complex shapes and becomes strong as it hardens, making it adaptable to diverse bone defects. Under physiologic conditions, CAP erodes before the slower-degrading CA matrix, enabling a dynamic system that generates interconnected pores and tunable growth factor release profiles and degradation. A CA/CAP/NC composite allows flexible incorporation of multiple bioactive factors for varied effects: within CA for early, sustained release; within CAP for pulsed release; and/or into NC embedded within the CA/CAP for delayed, sustained release. This also allows factors to be released in parallel and/or sequentially. Detailed, long-term in vitro and in vivo characterizations of this cellulose biomaterial, including its ability to balance strength and porosity and the effects of osteoclasts on its degradation, remain knowledge gaps for advancing this transformative and natural biomaterial platform. Based on current knowledge, it is hypothesized that this dynamic cellulose-based putty will impart composition-dependent changes of strength and erosion in 3D microenvironments leading to varied bioactive factor release rates, vasculature development, and tissue ingrowth during bone repair. This will be tested in four Specific Aims: Aim 1: Characterize physicochemical and release properties of novel cellulose derivatives and compositions in vitro. Aim 2: Evaluate biocompatibility and bioactivity of released molecules in an in vivo subcutaneous implantation model. Aim 3: Evaluate cellular effects of putty formulations with early to long-term release profiles on a cranial flat-bone healing defect. Aim 4: Assess putty formulations with early to long-term release profiles on bone healing at a load- bearing site in a critical-sized long-bone defect in rabbit ulna. These studies will address several knowledge gaps for using cellulose biomaterials in bone healing. If this enabling putty technology is successful, it may be transformative to the field and adapted for other repair challenges in bone as well as a coating for biomedical implants.

NIH Research Projects · FY 2026 · 2025-04

Abstract: Despite the advent of combination antiretroviral therapy (cART), HIV-associated neurocognitive disorders (or NeuroHIV) remain prevalent, affecting 30-50% of patients and presenting persistent challenges in managing neurological complications. Glial cell dysregulation, particularly microglia and astrocytes, drives chronic neuroinflammation in the central nervous system (CNS), exacerbated by HIV proteins such as the Transactivator of Transcription (Tat). While previous research has implicated inflammasomes, notably the NLRP3 inflammasome, and toll-like receptors (TLRs) in mediating neuroinflammation, the precise mechanisms by which HIV, cART, and substances like cocaine modulate inflammasome activation and subsequent neuroinflammatory responses remain poorly understood. Also, pyroptosis, a highly inflammatory form of programmed cell death, emerges as crucial in inflammatory diseases, including NeuroHIV. Characterized by membrane pore formation and the release of proinflammatory cytokines like interleukin-1β (IL-1β) and IL-18, pyroptosis exacerbates inflammation and tissue damage. However, the link between inflammasome-mediated pyroptosis and microglial activation in the context of HIV Tat, cocaine, and cART, contributing to neuroinflammation in NeuroHIV, remains elusive and forms the crux of this proposal. This proposed study aims to address these critical knowledge gaps by elucidating the molecular mechanisms underlying the interplay between HIV Tat, cocaine, and cART in activating NLRP3 inflammasome signaling pathways, leading to pyroptosis and microglial activation. We hypothesize that these factors collectively contribute to microglial activation and exacerbate neuroinflammation through distinct regulatory mechanisms called microglial pyroptosis. This hypothesis will be tested with two specific aims: 1. determining the molecular mechanism(s) underlying NLRP3-mediated pyroptosis in microglia exposed to HIV Tat, cocaine, and cART in vitro; 2. validating the combined effects of HIV Tat, cocaine, and cART on NLRP3-mediated pyroptosis and microglial activation in vivo and isolated ex vivo adult microglia. Archival macaque brain tissues will also be utilized to validate the microglial pyroptosis and neuroinflammation. This comprehensive investigation into the molecular mechanisms of microglial pyroptosis and neuroinflammation induced by HIV Tat, cocaine, and cART holds significant promise for developing novel therapeutic strategies to alleviate neuroinflammatory responses in NeuroHIV and substance abuse disorders. Furthermore, the identification of these pathways may uncover new drug targets for the treatment of other neuroinflammatory diseases, underscoring the broader implications of this research.

NIH Research Projects · FY 2026 · 2025-04

ABSTRACT The primary objective of this proposal is to investigate Cocaine/HIV/SIV induced changes in inflammasome activation and their epigenetic regulation in the intestine and brain and its modulation by long-term, low-dose cannabinoids and combination antiretroviral therapy. The NLRP3 (NOD-like receptor family, pyrin domain containing) inflammasome is a multiprotein complex that elicits an innate immune response to infection and cell stress through activation of caspase-1 and processing of pro-interleukin-1β (pro-IL-1β) and pro-IL-18 to the bioactive IL-1β and IL18, respectively. Cocaine induces expression of inflammasomes and their associated genes in chronic HIV/SIV infection leading to persistent CNS and gastrointestinal tract (GIT) inflammation, which contributes to microbial translocation, immune activation, and HIV disease progression. Nevertheless, the epigenetic regulation of HIV/SIV and cocaine-induced inflammasome activation and its modulation remains unknown and needs to the investigated to better understand disease pathogenesis. Therefore, here we propose that supplementing 9-THC along with cART may reduce cocaine-induced inflammasome activation, dysbiosis, restore immune function, and slow HIV/SIV disease progression by modulating DNA methylation and gene expression. Our preliminary data showed marked to significant upregulation of caspase-1, gasdermin and NLRP6 inflammasome gene expression in basal ganglia (BG) and colonic epithelium (CE), respectively of chronically SIV-infected rhesus macaques. Interestingly, increased NLRP6 gene expression in CE was accompanied by extensive CpG promoter hypomethylation of up to 12 CpG sites. More importantly, long-term 9-THC administration significantly reduced CASP1 and IFN stimulated gene expression in BG and NLRP6 DNA hypomethylation in CE. Therefore, we will test the central hypothesis that a) aberrant DNA hypomethylation of inflammasomes and their associated genes in cocaine induced during chronic HIV/SIV infection results in their elevated expression leading to persistent CNS and GIT inflammation, which in turn contributes to microbial translocation, immune activation (microglial and T cells), and HIV disease progression; b) combining cART with low-dose of THC as a therapeutic may reduce cocaine-induced inflammasome activation, gut dysbiosis, and restore immune function. In order to test this hypothesis, we have proposed three specific aims. We will first investigate epigenetic mechanisms underlying cannabinoid-mediated reduction of cocaine-induced inflammasome activation in blood monocytes, brain, and CE during chronic SIV infection (aim 1). Next, we will determine the ability of THC administered in conjunction with cART to reverse cocaine-induced dysregulation of the microbiota-gut-brain axis (aim 2) and finally identify potential molecular mechanisms associated with THC counteraction of cocaine-induced DNA hypomethylation and inflammasome activation in in vitro cultured microglia, and CE cells (aim 3). The new findings from these studies will have important therapeutic implications for immune modulation in not only HIV but also other chronic intestinal inflammatory/neurodegenerative disease.

NIH Research Projects · FY 2026 · 2025-04

Sleep impairments are commonly reported in Fragile X syndrome (FXS) and autism spectrum disorders (ASD). Sleep impairments can have profound negative impact on brain development, cognition and general well-being however the underlying brain defects of sleep impairments in FXS are not well understood. While most research of sleep mechanisms have focused on neurons, recent studies revealed a critical role of astrocytes, a type of glia in the brain, as regulators of sleep and wake. Specifically, astrocyte Ca2+ signals exhibit distinct features across the sleep-wake cycle, are reduced during sleep compared to wakefulness and manipulating astrocyte Ca2+ signaling impacts sleep. Astrocytes in FXS have hyperexcitable Ca2+ signaling in culture and our preliminary data demonstrate the same in vivo. The goal of this proposal is to investigate the contribution of astrocytes and especially increased astrocyte Ca2+ signaling, to sleep impairments in FXS. We will ask the following important questions: How is sleep architecture impaired in FXS mouse models across development and adulthood and what is the contribution of astrocytes? In Aim 1 we will perform longitudinal telemetry to measure brain rhythms and behavior to determine the sleep architecture in the Fmr1 KO mice. To identify the contribution of astrocytes we will also record from astrocyte specific Fmr1 deletion (cKO) and restoration (cON) mice. Next, we will ask how is astrocyte Ca2+ signaling altered during sleep-wake cycle in FXS mouse models? In Aim 2 we will record cortical astrocyte Ca2+ activity with 2-photon microscopy in awake and naturally sleeping mice while monitoring brain rhythms and behavior. We will also image the dynamics of norepinephrine (NE) and NE-projections in the cortex. Finally, in Aim 3 we will determine the causal relationship between altered astrocyte Ca2+ signaling and sleep impairments using genetic and chemogenetic approaches and determine the impact on sleep- dependent synaptic plasticity. The approach is intellectually and technically innovative as a first study to link FXS sleep deficiency and altered cortical astrocyte Ca2+ activity and because it employs a novel combination of state-of-the-art approaches. The proposed research is significant because it is expected to provide critical knowledge of the molecular and cellular mechanisms by which sleep impairments are regulated in FXS. Ultimately, such knowledge is expected to guide the development of astrocyte-specific therapies for treatment of sleep impairments in FXS and autism spectrum disorder.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY Chemotherapy-induced neuropathic pain (CINP) is a common side effect for cancer patients and can greatly reduce patients’ quality of life. Although a wide range of pharmacological agents have been investigated in clinical trials as potential CINP therapies, currently approved analgesics for neuropathic pain are inadequate and offer limited beneficial effects in ameliorating CINP. Flupirtine, a non-opioid and non-steroidal anti- inflammatory drug analgesic, can effectively relieve various pain conditions in clinical studies. However, safety concerns, particularly rare but severe liver toxicity, led to restrictions on its use and eventual withdrawal from the market in the U.S. The adverse side effects of flupirtine that induce hepatotoxicity have been shown to not be linked with the primary antinociceptive mechanism of action. We have developed a flupirtine analogue library and demonstrated that one of the compounds (compound 5) had a much faster metabolism rate than the flupirtine parent and did not induce hepatotoxicity after intraperitoneal injection. We have also successfully developed both short-term and long-term mouse CINP models. Our results showed that compound 5 effectively ameliorated short-term CINP, as evidenced by various pain behavior tests. In this proposal, we will further synthesize and screen flupirtine analogues through structural modification to enhance their liver safety profile while maintaining or even improving therapeutic efficacy to effectively relieve CIPN. The specific aims of the studies are (1) to strategically develop potent Kv7.2/7.3 activators unable to form hepatotoxic metabolites, (2) to determine the analgesic efficacies of flupirtine analogues in the CINP mouse models. This proposal will develop a series of novel flupirtine analogues with reduced or eliminated hepatotoxicity as non-opioid analgesics to relieve CINP and treat various other pain conditions and non-pain-related diseases.

NIH Research Projects · FY 2025 · 2025-02

Parkinson’s disease (PD) results from progressive loss of substantia nigra dopaminergic cells and manifests clinically as impaired motor control. Non-motor symptoms are far more disabling for patients, precede the onset of motor symptoms by a decade or more, are more insidious in onset and less effectively treated. Sleep dysfunction is often the most burdensome of non-motor symptoms, is pervasive in PD patients, and includes sleep fragmentation, insomnia, excessive daytime sleepiness, and rapid eye movement (REM) behavior disorder. Changes in sleep macro-architecture are also seen in PD, with less total sleep time, increased wake after sleep onset (WASO), increased non-REM stage 1 (NREM1) sleep, and decreased NREM2, NREM3, and REM. Building on our earlier observations of spectral patterns in subthalamic nucleus local field potentials (STN LFP) that correspond to specific sleep stages, we have used a novel investigational DBS programmable generator (RC+S Summit System; Medtronic) to enable exploration of sleep biomarkers and prototyping of therapeutic closed-loop, stimulation (DBS) algorithms. Specifically, in PD patients undergoing STN DBS, we examined whether STN oscillations correlated with sleep-stage transitions, then constructed and evaluated sensing and adaptive stimulation (aDBS) paradigms that allow ongoing sleep-stage identification, and attempted to induce through aDBS an increase in sleep-stage duration associated with restorative sleep. This work addresses an unmet clinical need—i.e., the significant sleep dysfunction of PD—and enabled evaluation of STN aDBS in PD patients, specifically for the treatment of sleep dysfunction. Our primary hypothesis for UH3NS113768A1 was that STN—a highly interconnected basal ganglia node—affects the regulation and disruption of human sleep behavior and may be modulated for therapeutic advantage. We tested whether dysfunctional PD STN activity correlates with sleep fragmentation, and whether STN aDBS algorithms could be developed that improve sleep-stage maintenance and sleep quality. Results demonstrate that PD sleep disturbance is marked by pronounced fragmentation and under-expression of NREM3 and REM, reinforcing prior reports of disrupted macroarchitecture. Sub-clinical STN DBS did not alter overall sleep-stage expression, though effects on sleep spindle density support microarchitectural benefits. Efforts to implement closed-loop aDBS for sleep in the home environment revealed practical challenges, including wearable limitations and subject compliance, but also highlighted how next-generation DBS devices with enhanced recording and user interface capabilities can empower novel aDBS strategies. While these findings do not yet confirm a definitive approach to restoring sleep in PD, they underscore that STN-LFP signals hold promise as biomarkers for identifying and modulating specific sleep states in real-world settings and set the stage for more targeted, long-term interventions to improve sleep quality and potentially slow disease progression. Progress on study Aims 1 and 2 described above was impeded by the COVID Pandemic, and has gathered steam with navigation past obstacles to patient recruitment and surgeries. Enrollment for Aim 1 and 2 and investigation of Aim 1 subjects are complete, and the 3-week in-home sleep trial for Aim 2 subjects will conclude prior to Year-5 end. We recently developed novel methods for analyzing study data and are requesting additional funding to accomplish this.

NIH Research Projects · FY 2026 · 2025-02

PROJECT SUMMARY Staphylococcus aureus is a gram-positive bacterium that colonizes the nares of ~30% of the human population and is the most common cause of bacteremia, endocarditis, and skin and soft tissue infections. Environmental nitrogen acquisition is essential for bacterial growth. To meet intracellular nitrogen requirements, S. aureus requires the transport of exogenous nitrogen via cytosolic NH4+ ions, amino acids, nitrate, nitrite, or urea. A common nitrogen source for many pathogens is glutamine, which along with glutamate function as the primary amino donors for cellular reactions requiring nitrogen. Since many precursor molecules for glutamine synthesis are also essential for other intracellular reactions, these pathways are tightly regulated processes affected by intracellular glutamine concentrations. Human hosts have a high level of free glutamine, so we hypothesized that glutamine transport serves as the primary route for nitrogen acquisition in S. aureus. However, glutamine transport alone does not cause a virulence defect in S. aureus. We therefore hypothesize that S. aureus uses preferred nitrogen sources and that intracellular glutamate and NH4+ pools are regulated by GltC and GlnR, respectively. This hypothesis will be addressed by two experimental aims. In aim 1, we will use 15N- isotopolog tracing experiments with LC-MS to identify preferred nitrogen sources in aerobic, microaerobic, and anaerobic conditions. Additionally, we will use both a bacteremia and skin soft tissue model of infection to determine if nitrogen limitation results in a physiological phenotype. In aim 2 we will use a gel-shift assay to characterize the DNA binding activity of the nitrogen metabolic regulator GlnR both individually and in the feedback inhibited glutamine synthetase (FBI-GS) complex with glutamine. We will also identify and evaluate the function of the regulatory GltC using qRT-PCR and RNAseq. This project seeks to research the unexplored role of nitrogen metabolism and regulation in S. aureus pathogenesis. Additionally, these data will inform systems biology approaches to determine novel therapeutic approaches to inhibit staphylococcal disease. The knowledge and training that I will gain through the experiments and activities proposed in this F30 application will provide me a comprehensive background in bacterial genetics and metabolism. This will be essential for my career as an infectious disease physician scientist as it will provide me the background needed to understand clinical genetics and metabolomics reports as our ability to characterize both pathogens and the microbiome expands. My laboratory is part of the Center for Staphylococcal Research (CSR) at the University of Nebraska Medical Center which fosters collaboration between faculty studying S. aureus metabolism, genetics, and host-pathogen response. Weekly meetings with members of the CSR will ensure that my understanding of S. aureus pathogenesis extends beyond the perspective of my laboratory. Weekly meetings with my mentor and mentorship from the senior scientists and junior faculty in my laboratory will ensure I have the proper support and training to excel in the field of prokaryotic biology.

NIH Research Projects · FY 2026 · 2025-01

Abstract While antiretroviral therapy (ART) has significantly reduced AIDS-related mortality, significant treatment gaps remain among children living with HIV-1 infection. The potential impact of emerging potent pediatric formulations on viral suppression rates has been limited by factors common to daily oral ART. Additionally, the pharmacokinetic (PK) profiles of orally administered drugs in children is dynamic making drug delivery by this route unpredictable. Therefore, long-acting (LA) non-oral drug delivery approaches that could potentially overcome these challenges are of significance. We have created a 6-month aqueous injectable formulation of a bictegravir (BIC) prodrug with a shorter PK tail. The nanoformulated esterified medicine referred to as NDBIC sustains plasma BIC concentrations of > 4X the 95% inhibitory concentration after a single injection. Based on these encouraging results, we seek funds to launch an innovative approach that will produce a complete every 6-months integrase strand transfer inhibitor (INSTI) based regimen for treatment of HIV-1 infected pediatric patients. Our innovative approach is focused on transforming well established daily oral pediatric therapies; BIC with dolutegravir as the alternative INSTI, emtricitabine (FTC) and tenofovir (TFV) prodrugs, into ultra- long-acting (ULA) dose flexible prodrug formulations. Pediatric friendly formulation excipients, prodrug linkages and aqueous buffers will be used to further de-risk the process. We will leverage on the expertise of our multidisciplinary team with a strong history of collaboration, pharmaceutical industry support, animal models of pediatric drug development and PK/PD modeling to identify a safe and efficacious complete prescription ULA- ART formulation. Our approach is supported by extensive and rigorous preliminary data. Drug choices, prodrug, and formulation development, in vitro and in vivo screening and mechanisms, toxicology, PK, and pharmacodynamic (PD) profiles in relevant animal models of pediatric drug development (Sprague Dawley rats, humanized mice and non-human primates) will follow thoughtfully well-designed milestone-driven Go-No Go criteria that seek to address the unmet needs of the pediatric patient, simplify manufacturing, scale up and storage, evaluate potential adverse reactions of non-oral ART, and clinical pharmacology. We posit that sustaining therapeutic drug levels in viral target cells (monocyte-macrophages and CD4+ T cells) and tissues will improve treatment outcomes. A twice/year complete pediatric ART regimen will be identified for future clinical development following the guidance and recommendations by the US FDA.

NIH Research Projects · FY 2025 · 2025-01

Project Summary/Abstract The goal of this project is to develop nanostructures that are selective for bacteria and to create a theoretical framework (in silico model) that can be used to predict the activity of nanostructures and their behavior and interactions with bacteria. Bacterial infections are a serious health problem that affects the human population. Further, the emergence of resistant bacterial strains has made the problem even more serious. Two pathogens of particular concern are Staphylococcus aureus and Acinetobacter baumannii. These pathogens are a main cause of nosocomial skin and lung infections and are prevalent in immunocompromised patients and hospital settings. To deal with these serious pathogens we propose to create nanostructures with intrinsic antibacterial activity that can also target the bacteria by including siderophore molecules. In addition, we propose to develop theoretical (computer) models that can be used to understand the biological action of the nanostructures and can be used to design new nanostructures. To develop these systems, we have devised four specific aims. Specific Aim 1. Computer-Guided Design of PA nanostructures. We will design several PA molecules and use theoretical tools to select the PAs to be prepared. We will prepare nanostructures and test their antimicrobial activity and cell toxicity. Specific Aim 2. Developing Bacteria Targeting Nanostructures. We will modify the best PA molecules with siderophores. We will prepare nanostructures and test them against diverse bacteria. We will also use the data to refine our MOLT system Specific Aim 3. Membrane Interactions and in Vivo Activity. We will study the ability of nanostructures to disrupt membranes. We will also study the role of the siderophores. Finally, we will evaluate their antibacterial activity using a Wax moth larvae in vivo model. Specific Aim 4. Developing a Multi-Resolution Model model. We will combine the statistical mechanics and molecular dynamic simulations to predict and understand the shape of the nanostructures, their activity. We will also study the interactions of the PA nanostructures with membranes using both models. We will then combine the models to get a better picture of the membrane disruption process. We will use the model to develop Quantitative Structure-Activity Relationships that can be used to make better nanostructures.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY/ABSTRACT Medulloblastoma (MB) is the leading cause of cancer-related mortality in childhood. Divided into four subgroups, patients with group 3 tumors (G3MB) have the lowest survival rate (<50% at 5 years). This has been attributed to a lack of targeted therapies and high-risk features, like deletions along the short arm of chromosome 17. These deletions target critical tumor suppressor genes like miR-1253, a brain-enriched microRNA active during cerebellar development. Restoring miR-1253 expression in G3MB cell lines reduced cancer cell growth and aggressiveness, inhibiting several oncoproteins, including B7-H3, which has been strongly linked with metastasis and immune evasion in many aggressive cancers. Silencing B7-H3 expression in G3MB cells dramatically reduced their growth and invasiveness, as well as the expression of SLUG, a JAK2/STAT3 signaling product associated with metastasis. This kindled our interest in finding drug compounds to silence B7-H3 signaling as a novel way to treat G3MB tumors. By screening an extensive collection of molecules known to interact with B7-H3, one highly potent compound, B7-H3-Ni1 (Ni1), was identified, which inhibited G3MB cancer cell viability at low micromolar concentrations. The overall objective of this proposal is to demonstrate a mechanistic proof-of-concept for B7- H3 inhibition as a therapeutic approach against G3MB. We hypothesize that B7-H3-Ni1 can suppress G3MB tumor growth and aggressiveness by activating immune cell function and inhibiting JAK/STAT signaling pathways associated with metastasis. In Aim 1, we will map the molecular binding of Ni1 to B7-H3. For immune activation, a syngeneic G3MB cell line (MP-1) co-cultured with mouse Tregs, CTLs, or NK cells will be treated with vehicle, Ni1, or si-B7-H3 to study the effect of B7-H3 blockade on immune cell degranulation, target cell lysis, cytokine production, and cancer cell viability. For JAK2/STAT3 signaling, we will measure the impact of vehicle vs. Ni1 vs. si-B7-H3 on metastasis- associated signaling partners of B7-H3, metastasis-associated effectors of JAK2/STAT3 signaling, and apoptotic markers. Impact on cancer cell growth, viability, and invasiveness will also be assessed in vitro. Then, we will examine if Ni1 signaling is enhanced by a STAT3 inhibitor (WP1066). A B7-H3 knockout (B7-H3KO) G3MB cell line will be incorporated into all in vitro studies. In Aim 2, an immunocompetent G3MB mouse model that most closely recapitulates human disease will be used to illustrate the immunostimulatory and anti-neoplastic properties of Ni1. We expect to demonstrate that treatment with Ni mitigates tumor burden and longevity by downregulating JAK2/STAT3-associated metastasis signaling. At completion, our studies will address a critical gap for G3MB patients who suffer disproportionately high mortality rates amongst MB patients by characterizing a novel anti-tumor agent that may be efficacious and well-tolerated. By identifying a new druggable target (B7- H3), we may unlock the potential to transform the treatment of advanced MB tumors.

NIH Research Projects · FY 2026 · 2025-01

Project Summary Light responses generated by photon capture in rod photoreceptor cells must be transmitted at their synapses to downstream neurons. Transmission at the first synapse in the retina thus determines the information about light available to the visual system. A remarkable ability of the human visual system is the ability to detect dim light flashes consisting of only a few photons. This sensitivity originates with the ability of rods to respond to single photons but requires nearly noiseless transmission of small single photon responses through the retina and brain. A critical first step in this process is transmission from rods to rod bipolar cells. Fluorescent glutamate sensors have provided important insights into circuit mechanisms in other parts of the retina and CNS but have not been successfully used to study release from rods. To overcome this, we developed a genetically-modified mouse line allowing constitutive expression of an improved glutamate sensor, iGluSnFr3, specifically in rods. In this proposal, we plan to characterize the strengths and limitations of using iGluSnFr3 to measure glutamate at rod synapses. Electrophysiological recordings from tiny mammalian rods are exceptionally challenging so providing an optical approach for studying glutamate release from rods will pave the way for future work by many laboratories on basic and disease-related questions of photoreceptor function. Release rates at rod synapses are slow and this makes it difficult for a post-synaptic rod bipolar cell to discriminate a lengthy random pause in release from a genuinely lengthy pause caused by a single photon response. One way to overcome this would be for release to occur at regular, not random, intervals. Results from our laboratory using an indirect measure of glutamate release suggested that synaptic release from rods is surprisingly regular, not random as generally thought. Optical measurements using iGluSnFr3 offer the opportunity to directly measure release from dark-adapted rods in intact retina and test the physiological significance of this regularity. Establishing that release from rods occurs at regular intervals would be a fundamental change in our understanding of how visual information is encoded at the first synapse in the retina and suggest many future experiments.

NIH Research Projects · FY 2026 · 2024-12

Porcine Platforms For Technology Development In Pancreatic Cancer PROJECT SUMMARY/ABSTRACT The long-term goal of this research is to develop a platform on which experimental therapies and devices for pancreatic cancer (PC) management can be advanced to the clinic in a more efficient manner than achievable with current preclinical PC models. The focused objective of this R01 application is to demonstrate the utility of a transgenic porcine model of cancer (the KRAS/p53 Oncopig) in testing the antitumor efficacy of novel neoadjuvant therapies and also in testing the effectiveness of three endoscopic medical devices for imaging and treatment of PC. Murine PC models may not adequately reflect human tumor biology because of differences in size, physiology, anatomy, and genetic sequence with humans. So, the rationale to use a porcine model of PC is that it should be more predictive of human tumor biology and response to therapy than murine models are. This R01 application will particularly use the swine's size advantage. The hypothesis of this R01 application is that transgenic porcine models of PC are feasible and useful for testing of PC applications. In Aim 1, the ability of the transgenic porcine PC model to replicate the entire neoadjuvant therapy/surgery/survival sequence of human PC management will be demonstrated. In Aim 2, two novel endoscopic technologies for imaging and treatment of PC will be tested in the Oncopig, in studies which would not be feasible in a mouse. The work proposed in this R01 application will be accomplished through a collaborative, multi-faceted team that will include a general-oncologic surgeon (PI), molecular/cellular biologists with expertise in PC and gene editing, a medical oncologist who manages patients with PC, a pathologist specializing in pancreatic/GI cancers, sequencing and bioinformatics experts, a radiologist focused on MRI research, and a biostatistician. This project is innovative because the type of models proposed and the novel technologies that will be tested with these models. The impact of validated porcine models of PC would be to enhance, complement, and supplement preclinical data from other tumor models, and also to advance experimental technologies for PC management to the clinic in a more efficient manner, with fewer experimental therapies failing in clinical trials.

NIH Research Projects · FY 2026 · 2024-12

PROJECT ABSTRACT Alcohol use disorder (AUD) rates have doubled in the past five years. Despite having effective treatments, over 50% of patients relapse within the first year. Theories suggest that a primary reason for high relapse rates is the development of anxiety during abstinence that causes continued alcohol use. That is, individuals with AUD use alcohol for its negative reinforcing property, which removes anxiety that occurs during alcohol abstinence. Chronic alcohol use impacts negative reinforcement neural circuits, which contain the bed nucleus of the stria terminalis (BNST) at its core. The BNST is a region that contributes to negative affect, mediates stress-induced reinstatement (relapse), and was recently implicated in negative reinforcement. The BNST has extensive inputs from regions implicated in reward and emotion processing—specifically, the amygdala, ventral and dorsal striatum, and dorsal anterior cingulate cortex (dACC). In alcohol research, the primary emphasis has been on positive reinforcers (i.e., alcohol), providing us with extensive knowledge about the effects of positive reinforcement on behavior and brain function in AUD. In contrast, the alcohol field has a limited understanding of the effects of negative reinforcement on behavior and brain function. Considering that conceptual models of AUD cite negative reinforcement as a driver of relapse, this represents a major gap in knowledge. Human studies can leverage findings from animal models to provide a better understanding of negative reinforcement in AUD. Specifically, cross-species translation will be foundational for determining underlying behavioral and brain mechanisms of AUD and relapse. To address cross-species translation and underlying mechanisms of negative reinforcement, we formed a collaboration with a preclinical researcher to translate a well-validated animal paradigm of negative reinforcement into humans. The proposed project will investigate negative reinforcement behaviors and connectivity in a negative reinforcement neural network in adults with AUD who are in early abstinence (EA). Specific Aim 1 will characterize negative reinforcement learning and bias in EA adults compared to controls (CON). Specific Aim 2 will determine connectivity strength of a BNST negative reinforcement neural network using intrinsic (‘resting state’) functional connectivity and diffusion tensor imaging (DTI). Both aims will test for group differences (EA, CON) and exploratory analyses will determine whether negative reinforcement network connectivity is associated with negative reinforcement behaviors in the task. The hypotheses are that EA participants will (1) learn negative reinforcement faster and have a bias for negative reinforcement and (2) display stronger negative reinforcement network connectivity than controls. The results of this study will fill a critical gap in knowledge to better understand the behavioral and neural mechanisms that underly AUD and relapse, representing a critical target to guide future prevention and treatment of AUDs.

NIH Research Projects · FY 2025 · 2024-09

Abstract: Alcohol consumption adversely affects up to 28% of hospitalized patients and contributes to a loss of 133 million disability-adjusted life years and 5.3% of worldwide deaths each year. As alcohol use has substantial health and economic impact, much attention has been directed toward the numerous adverse health outcomes in patients with unhealthy alcohol use. Alcohol use disorder (AUD) occurs on a spectrum from mild to severe and is precipitated by binge drinking and heavy alcohol use. We and others have shown that AUD is a modifiable perioperative risk factor and is present in up to 18% of surgical patients. Despite its significant clinical impact, AUD is often overlooked in the design of perioperative care plans. Although AUD affects 9% of the US population, less than one in ten individuals with AUD receives any treatment. Barriers to treatment are multiple and include scarce care availability, limited access, and social stigma. Although anesthesiologists routinely provide guidelineconcordant treatment for non-operative medical conditions such as coronary artery disease, AUD-specific care is rarely provided, even to high-risk patients. Thus, there is a compelling opportunity to integrate AUD screening and treatment into routine perioperative care. Our central hypotheses are that AUD health services initiated in the perioperative period will 1) leverage the significant resources made available to perioperative care in the US, 2) forge novel synergistic alliances between previously disconnected healthcare settings, and 3) break down barriers to access to care. A multi-institutional team of anesthesiologists, biostatisticians, psychologists, and psychiatrists with expertise in AUD treatment and novel clinical trial designs will lead the Leveraging Alcohol Use Disorder Screening for Treatment in Routine Perioperative Care: AllUsCare proposal and proposes three aims: 1) Leverage our existing EHR-integrated Alcohol Use Disorders Identification Test-Concise (AUDIT-C) screening tool for provision of AUD-specific perioperative care, 2) Conduct a two-center prospective observational cohort study to assess patient acceptability of interventions, feasibility of outcome data collection, and optimum outcome measures for a future pragmatic trial, and 3) Optimize perioperative AUD intervention bundles most likely to be effective in a future pragmatic randomized factorial cluster trial. This R34 planning grant will lay the groundwork for identifying the most effective health service intervention bundles in surgical patients at high risk for AUD. To maximize generalizability to other populations, we will conduct our study in two centers that serve inner-city and rural populations. At the conclusion of AllUsCare, we will have established the research team, designed an AUD intervention bundle most likely to be effective in a future multi-center pragmatic trial, demonstrated a single-IRB governed uniform data collection and entry process, and confirmed the acceptability and feasibility of the future pragmatic trial. This application directly responds to NIAAA FOA #PAR-22-157.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Despite decades of research documenting the benefits of continuous positive airway pressure (CPAP) therapy, this highly effective treatment is not initiated, implemented, or persistently used by the millions of adults worldwide diagnosed with obstructive sleep apnea (OSA). An estimated 16% of patients refuse CPAP after a single night’s use, 29-83% do not adhere to current CPAP recommendations, and up to 50% abandon CPAP after one year. Untreated OSA is associated with a range of distressing symptoms, dangerous comorbid conditions, billions in healthcare spending, and higher all-cause mortality. Growing evidence shows that older adults, people with low socioeconomic status, and certain racial/ethnic minority groups are at higher risk for developing OSA, less likely to adhere to CPAP therapy, and experience worse outcomes than other groups. Other variables associated with CPAP non-adherence are commonly experienced CPAP side effects, knowledge deficits, low levels of self-efficacy, motivation, and confidence, and fragmented healthcare delivery. A variety of educational, supportive, behavioral, and mixed interventions have been tested in randomized controlled trials (RCTs). While each of these intervention types has demonstrated efficacy to varying degrees, key knowledge gaps remain as to: (1) which intervention type leads to the greatest improvements in long-term CPAP adherence and clinical outcomes; (2) who benefits most from a given intervention; and (3) what duration and/or sequence of interventions is required to effectively induce behavior change. Collectively, these gaps suggest the need for the development and testing of adaptive interventions aimed at improving CPAP adherence. Adaptive interventions capitalize on differences in how patients respond to intervention by providing appropriate modifications for those who need them and when they need them. To our knowledge, no fully powered RCTs have tested the effectiveness of adaptive interventions aimed at increasing CPAP adherence. Project objective: is to evaluate a telehealth-delivered adaptive intervention for persons newly diagnosed with OSA that promotes CPAP adherence and improves health outcomes and assess potential mediators and moderators of treatment response. Aim 1: Compare effects of motivational enhancement, device support, and standard care on primary and secondary outcomes at 3 mo. Aim 2: Determine which behavioral change treatment is most efficacious at promoting primary and secondary outcomes in participants who respond and do not respond to initial treatment at 6 and 12 mo. Aim 3. Assess if CPAP adherence is mediated by stage of change, self-efficacy, knowledge, and/or confidence and moderated by social economic status, race, and sex at 3 mo. By using an innovative adaptive design and telehealth-delivered approach, study results will directly lead to a more personalized approach to increasing long-term CPAP adherence and improving health outcomes of persons with OSA while simultaneously generating a better understanding of factors that drive CPAP adherence across a variety of personal, social, and economic contextual factors.

NIH Research Projects · FY 2025 · 2024-09

Alzheimer's disease (AD) is a pressing issue in the United States, affecting over 6 million Americans and ranking as the fifth-leading cause of death in 2020 for persons 65 and over. Approximately 90% of AD cases are sporadic, reflecting an established contribution to AD risk of environmental and lifestyle risk factors (ERF). ERF impact AD risk throughout the lifespan, but ERF during critical developmental periods may influence late- life disease risk beyond what is currently appreciated. This may be demonstrated by abnormal brain maturation (structural or functional) due to ERF exposure. While late onset AD is not often diagnosed until one’s mid 60s, utilizing a lifespan approach acknowledges early life ERF that may influence risk to disease. Utilizing an AD lens can help investigate these early life influences, with the goal of identifying early interventions for reducing neurodegenerative risk. The F99 phase of my proposal will investigate prenatal and childhood ERF and their association with memory. The central hypothesis is that AD associated ERF (AD-ERF) during childhood are linked to the neurodevelopment of AD associated brain systems (hippocampus, hippocampal dependent memory), and these links affect AD vulnerability in late life. There is a notable gap in understanding how early- life AD-ERF bias cognitive development towards neurodegenerative risk. To investigate this, I will measure how a person’s environmental exposures (stress, socioeconomic status, neighborhood deprivation index) in the prenatal, personal, and parental environments (as identified through a factor analysis using the Adolescent Brain and Cognitive Development study: Aim1A) controlling for familial AD risk (via grandparent/great grandparent blood biomarker assay: collected from parent study) influence hippocampal dependent RM in children. The F99 phase will include training in reproducible neuroimaging, advanced statistics, and scientific communication. Completion of the F99 phase lays a strong intellectual, technical, and professional foundation for the postdoctoral (K00) phase of this award. During the K00 phase, training in epigenetics and AD- associated biological aging will develop knowledge, expertise, and skills essential to becoming an independent investigator. It is my long-term goal to establish a multi-disciplinary research laboratory focused on studying modifiable AD risk factors in order to develop and implement treatments or preventative programs that might reduce AD risk in populations with high vulnerability due to their environment. This research will shed light on the relationship between environments and development, offering insights into the predictive power of the environment in shaping AD risk. Ultimately, this work could lead to early interventions for AD, addressing the looming public health crisis posed by an aging, AD-vulnerable US population.

NIH Research Projects · FY 2025 · 2024-09

Project Summary The leading cause of death from ages 1-44 years in the United States is traumatic injury, responsible for millions of hospital encounters at a cost of over $4 trillion annually. Unfortunately, outcomes in trauma patients who survive the initial injury have not seen the marked improvements observed in other fields of medicine recently, so more research is critical. It is known that a key driver of delayed mortality after injury is organ failure, where a marked inflammatory response and dynamic coagulopathy are both implicated. Approximately 1 in 4 patients who suffer from major trauma have a detectable coagulopathy on clinical coagulation testing, and even more have a perturbation in fibrinolysis, a process mediated by the protease plasmin. A major inflammatory signaling pathway in the blood circulation is called complement, a protease cascade that shares a number of cleavage specificities with the coagulation system with increasing recognition the two processes heavily influence one another. It logically follows that acute traumatic coagulopathy and inflammatory organ failure may therefore be related processes, with complement-mediated inflammation being a key link. In support of this, the extant literature has shown the degree of complement activation correlates with both organ failure and mortality, but the primary mechanism of complement activation after trauma remains unknown. Our preliminary results and prior work suggest that the observed complement activation may occur as a direct result of plasmin cleavage of complement proteins, where plasmin is known to be generated in large amounts in trauma patients with severe shock. In addition, we also know that after trauma the vascular endothelium sheds its glycocalyx into the circulation, which may serve to activate complement via the lectin pathway, which is a complement activation pathway that binds to specific glycans and causes robust complement activation. To further elucidate the mechanism of complement activation after trauma, we are now proposing 3 Aims. In Aim 1, we propose a focused protein and glycan biomarker investigation of plasma from human trauma patients to test whether plasmin generation or circulating endothelial glycocalyx molecules correlate with degree of complement activation. In Aim 2, we will use a human endothelial cell culture model designed to mimic traumatic shock to elucidate whether plasmin, endothelial glycocalyx molecules and the lectin pathway, or both are capable of generating robust complement activation in the absence of other complement pathways to delineate relative contributions and mechanism of activation. Finally, in Aim 3, we will directly investigate the role of complement in generating organ failure in trauma using a mouse model of trauma comparing mice deficient in complement proteins C3 and C5 with wild-type mice. The results of this Aim would highlight the potential role of anti-complement therapies for future clinical study considerations. Taken together, this research program will improve our understanding of the mechanisms of complement activation and inflammatory organ injury after trauma in this leading cause of death in the United States.

NIH Research Projects · FY 2024 · 2024-09

Abstract Aging is associated with persistent levels of inflammation, which is also a hallmark of HIV infection. The effects of aging are particularly important to the population of people living with HIV (PLWH), as the overall cohort of infected individuals is getting older, with over 4 million over the age of 50. The reported effects of aging seen in HIV patients are increased co-morbidities, frailty, diabetes, cardiovascular disease, increased immune inflammation, and increased senescence, which led researchers to believe that HIV infection leads to a form of early aging. Though, the widespread use of antiretroviral therapy (ART) allowed the disease to be medically manageable, however, strict lifelong adherence to daily drug regimens is required. Though daily combinatorial ART intake keeps HIV-1 at bay in tissue reservoirs, replication-competent integrated proviral DNA can persist for decades as HIV-1 infected cells have an average half-life close to 44 months and could be responsible for accelerated aging. Also, despite all cART’s achievements, it’s poor penetrance across the blood-brain barrier leads to continuous low-level viral replication in the central nervous system (CNS) could be the cause of premature aging in PLWH. Thus, deciphering how much ART-mediated viral suppression affects aging and the brain immune homeostasis will provide clue to understand the mechanism involved in accelerated aging in PLWH and ultimately its therapeutic targeting. Defining the consequence of latent HIV-1 burden in the human brain during progressive aging is limited by sample access. We plan to answer this above question by deploying a small animal model of HIV-1 disease (NSG-humanized mice). Recently we employed NSG-humanized mice and demonstrated recovery of latent viruses from whole brain cells. Based on these findings and new preliminary data, we now propose to study progressive aging associated changes during HIV-infection in a long-term immune reconstituted humanized mouse model. How much HIV-1 latency and cART contribute to premature aging (we termed as LAAGING) and functional neuro-immune interaction at molecular and behavioral level will be answered. By employing the proposed animal model under highly suppressive cART treatment, the brain cells carrying the latent HIV-1 will be evaluated and characterized by transcriptomics and metabolic profiling. This will be followed by measurement of axonal fiber integrity using diffusion tensor imaging and behavior analysis to look for brain functional deficits during progressive aging. The outcome of the project will provide valuable information for the identification of new biomarkers of aging, assessment and monitoring of combination therapies for elimination of virus and its associated neuroinflammation in the CNS. Data from these ART- suppressed humanized mice brain studies will also provide the first-hand evidence of the existence of neuro- immune crosstalk both within and outside of CNS during progressive aging and molecular pathways being affected during HIV-latency and will help in future therapeutic targeting of CNS reservoirs in infected patients.