University Of Wisconsin-Madison

NIH Research Projects · FY 2026 · 2024-03

Local sleep and mental fatigue PROJECT SUMMARY Mental fatigue—a feeling of exhaustion often accompanied by the sense that every task requires great effort—is prevalent in neurological and psychiatric diseases. In healthy subjects, it is a common symptom ensuing after intense learning or sleep deprivation. Mental fatigue is also frequent when sleep is poor and can be relieved by improving sleep quality. Moreover, the broad cognitive impairment that accompanies mental fatigue is resolved by sleeping but not by resting while awake. Despite the evidence linking mental fatigue with brain dysfunctions and sleep deficits, the reasons why the brain gets “tired” remain unknown. This proposal tests a novel, circuit-level hypothesis about the neural mechanisms of mental fatigue caused by sleep loss or intense learning. In these cases, we hypothesize that a key underlying mechanism is the increasing occurrence, within corticothalamic networks, of local neuronal OFF periods in wake, triggered by the activation of Martinotti cells (MaCs). In NIH-funded groundwork that led to this hypothesis we demonstrated, in both rodents and humans, that sleep deprivation leads to “local sleep” in wake: even though the rest of the brain is awake, local groups of cortical neurons briefly stop firing (OFF periods), as they usually only do, in a widespread manner, during sleep slow waves. Even though subjects are behaviorally awake, if “local sleep” occurs in cortical areas involved in task execution, performance is impaired. In further work, we demonstrated that the widespread OFF periods underlying the slow waves of sleep are promoted by the activation of MaCs, which are somatostatin-positive (SOM+) GABAergic neurons found throughout cortex. Once recruited by strong excitatory inputs from pyramidal neurons, MaCs can act as master regulators of cortical excitability by inhibiting all other cell types. Finally, we found that extended wake and intense learning lead to increased neuronal excitability due to the progressive strengthening of cortical glutamatergic connections, which are renormalized (weakened) by sleep and not simply by rest. Guided by these findings and preliminary data, we propose that a key mechanism underlying mental fatigue is the increased excitability of MaCs and their maladaptive triggering of OFF periods during wake. To test this hypothesis, we will perform high-density Neuropixels recordings in mice and rats in multiple cortical and subcortical areas. These recordings will be combined with optogenetic tagging to determine, first, whether extended wake or intense learning will activate MaCs and trigger local OFF periods in wake. Guided by a new large-scale sleep/wake model of corticothalamic circuits, we will then test two key predictions in vivo. Using opto/chemogenetics, we will determine whether, after extended wake or intense learning, local OFF periods in wake are prevented by silencing MaCs and, conversely, whether they are triggered by the activation of MaCs even in fully rested animals. If successful, we will uncover: 1) a candidate circuit mediating mental fatigue after extended wake and intense learning; 2) a key mechanism through which fatigue is mediated—the induction of OFF periods in corticothalamic networks due to the progressive build-up of synaptic strength in wake. Exploratory analysis of unit data collected in thalamic, striatal, and hippocampal areas will also determine whether local sleep extends subcortically, potentially contributing to the impairment in performance and motivation associated with mental fatigue.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Mpox (formerly monkeypox) virus (MPXV) is a serious global health concern that recently caused a significant outbreak in the United States, resulting in over 30,000 infections. MPXV infections are known to cause high rates of fetal demise, particularly with clade I MPXV which is endemic in Central Africa. It is unclear whether clade IIb MPXV, which circulated worldwide and within the United States, also causes adverse pregnancy outcomes because the outbreak is so recent. Defining the rate of adverse pregnancy outcomes and whether vertical transmission occurs with clade IIb MPXV is a critical public global health challenge. In addition, it is uncertain whether the antiviral drug tecovirimat, currently used off label to treat mpox disease, can improve pregnancy outcomes when administered early in the course of infection during pregnancy. Determining whether early treatment with tecovirimat improves pregnancy outcomes is essential for informing clinical management. We developed a preclinical rhesus macaque model of prenatal Zika virus infection, defined vertical transmission pathways, and assessed adverse pregnancy outcomes. We will adapt this expertise to develop a preclinical rhesus macaque model of mpox disease in pregnancy. We hypothesize that MPXV will be vertically transmitted with inoculation in early gestation, that there will be an increase in adverse pregnancy outcomes with maternal infection, and that the antiviral drug tecovirimat will decrease maternal disease and improve pregnancy outcomes. We propose the following aims to test these hypotheses. Aim 1: To define the rate and pathways of vertical transmission following MPXV infection in early pregnancy. Aim 2: To define the association between maternal mpox disease severity and pregnancy outcomes following MPXV infection in early pregnancy. Aim 3: To determine if early antiviral treatment improves maternal mpox disease and pregnancy outcomes. Accomplishing these aims will fill a significant knowledge gap in our clinical management of pregnant persons infected with MPXV: whether infection results in a higher rate of adverse pregnancy outcomes, whether vertical transmission occurs, and whether early administration of tecovirimat improves maternal mpox disease and pregnancy outcomes. Given the limited clinical trial data on mpox in pregnant women, the findings from this study will be pivotal in informing future clinical trials and improving the clinical management of pregnant women with MPXV infection, leading to better health outcomes for both mothers and infants.

NIH Research Projects · FY 2026 · 2024-02

Human O-GlcNAcase (OGA) is the sole enzyme that hydrolyzes O-GlcNAcylation, an essential intracellular protein glycosylation that functionally regulates thousands of proteins in response to nutrients and stress. Aberrant functions of OGA have been detected in numerous diseases including cancer and neurodegeneration, fostering intense interest in OGA as a therapeutic target. However, due to the lack of complete OGA structure and the paucity of information on its substrate recognition, little is known about how OGA can accommodate such a broad range of substrates without a consensus sequence motif, while still maintaining certain level of specificity. Our preliminary studies provide strong evidence suggesting that OGA’s non-catalytic regions play critical roles in substrate binding and new functions that we are just beginning to reveal. We propose to advance structural and functional investigation of OGA. The new findings from this study will help decode OGA’s complex cellular regulation and will serve as a critical foundation for manipulating the precise role of OGA, paving the way for development of more effective and safer therapies for challenging diseases.

NIH Research Projects · FY 2026 · 2024-02

Project Summary/Abstract: The project addresses vascular contributions to cognitive impairment and dementia (VCID) using a design that enrolls persons with the autosomal dominant gene for Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL). Exploration of VCID by focusing on the rare heritable monogenic disorder, CADASIL, will advance knowledge of the full spectrum of this vascular dementia from asymptomatic gene carriers through dementia and will facilitate studies evaluating vascular contributions in different neurodegenerative processes including Alzheimer’s disease (AD) and related dementias (ADRD). CADASIL is caused by a mutation in the gene Notch3, which encodes a receptor protein expressed in vascular smooth muscle cells and pericytes. When mutated, expression of the gene leads to generalized degeneration of vascular smooth muscle cells affecting small and medium sized arteries. To provide insights into the impact of small vessel disease (SVD) in CADASIL (and AD, ADRD, and VCID more generally) on parenchymal damage and cognitive decline, non-invasive Magnetic Resonance Imaging (MRI) provides quantification of brain structure, function, and some measures of vascular health; however, current clinically available MRI techniques lack sensitivity and specificity to address key vascular hypotheses as many measures only indirectly study cerebrovascular disease, primarily focusing on the damage already caused by SVD, not the vessels themselves. Our group is uniquely positioned to address these key gaps in our knowledge base, both due to our MRI technology and as a member of the first CADASIL consortium study in North America. We propose an ensemble of novel MRI techniques optimized to provide vascular specific measures in a non-invasive imaging protocol. By characterizing in vivo measures of vascular stiffening at the macrovascular and microvascular levels in CADASIL, we will provide insights into the degree and mechanisms of VCID and establish baseline data to compare the longitudinal time course of CADASIL with normal age-related cerebrovascular changes, including SVD. We previously found that our MRI stiffness measures can detect vascular changes in amyloid positive AD participants even prior to demonstrating cognitive dysfunction. Our pilot data in CADASIL suggests pronounced vascular disease involving all levels of the cerebrovascular system. In close collaboration with three additional CADASIL consortium sites, a multidisciplinary team at the University of Wisconsin will first utilize newly developed, state- of-the art, quantitative MRI vascular imaging methods to understand the distribution and severity of vascular changes in CADASIL (Aim 1). We will then study the influence of altered intracranial vasculature on CADASIL neurodegeneration and cognition (Aim 2), and finally, we will study the degree of vascular involvement in CADASIL in comparison to cognitively unimpaired, normal aging participants (Aim 3). Upon completion we will be uniquely positioned to incorporate these techniques into large cohort studies investigating SVD, VCID, and extending into other ADRD vascular mechanisms as well as evaluate risk and protective factors.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY More than two million individuals are currently incarcerated in correctional facilities in the United States, and over half these individuals have a significant mental health problem. Despite the widespread need for mental health treatment in prisons, resources in this setting are extremely limited and most facilities are unable to provide adequate care. Hence, low-cost and scalable mental health treatment options are urgently needed for the prison population. Self-help workbooks are one such viable option. In non-incarcerated populations, self- help workbooks have shown efficacy in decreasing symptoms of depression, anxiety, and stress, as well as increasing mindfulness and acceptance skills. Moreover, our preliminary data on treatment preferences among incarcerated individuals indicate a preference for self-administered workbooks relative to other treatment formats (e.g., group or individual therapy with counselors). A self-help workbook for Acceptance and Commitment Therapy (ACT) may be particularly well suited to the prison environment. ACT aims to increase psychological flexibility, the ability to choose prosocial and value-based behavior, even if psychological barriers (e.g., anger, shame, maladaptive beliefs, etc.) are present. ACT self-help workbooks have been proven efficacious in reducing mental health burden in non-incarcerated populations. Moreover, studies of ACT group therapy in correctional settings have demonstrated feasibility as well as a variety of positive mental health outcomes. Hence, an ACT self-help workbook may be highly feasible, acceptable, and efficacious format for mental health treatment in the prison setting. The overall goal of this project is to conduct a pilot test of an ACT self-help workbook tailored specifically for the prison setting. Based on feedback from key stakeholders (incarcerated people and prison officials), we will make two main adaptations to the most widely used and empirically supported ACT workbook, in order to make it more acceptable and effective for the prison population: 1) reduce the reading level and 2) increase the personal relevance of the content (e.g., examples and exercises that are relevant to the prison setting). This project features a unique partnership between stakeholders (incarcerated individuals and corrections staff) and academic experts in forensic psychology and ACT. Our proposed research is directly responsive to Funding Opportunity Announcement (FOA) PAR-21-131, as it will adapt and test an intervention with demonstrated efficacy in a broader (and severely underserved) target population. As per the FOA, our study will include a pilot randomized controlled trial (RCT) to test the feasibility and acceptability of the intervention to inform a future larger-scale study. This study is novel in its use of stakeholder input to tailor a self-help intervention specifically for the prison setting. Successful completion of this pilot project will support a future larger-scale RCT evaluating the effectiveness of the workbook in prison. If proven clinically effective, widespread dissemination of the workbook could dramatically increase access to mental healthcare for a severely underserved population.

NIH Research Projects · FY 2026 · 2024-02

SUMMARY The proposed research focuses on processes that are mediated by proteins, that are crucial for cellular physiology and that are dysregulated in human disease. This effort has two components that differ topically but share common themes in experimental design. Fundamental mechanistic insight is our primary goal, but outcomes could support future therapeutic advances. We use chemically synthesized polypeptides to test molecular hypotheses, an approach that transcends the compositional limitations of biosynthetic polypeptides. One major focus involves signal transduction by a set of G protein-coupled receptors (GPCRs) that are naturally modulated by long polypeptide hormones (B family GPCRs). Agents that activate these receptors are used in human medicine. We conduct parallel studies with multiple receptors so that we can discern which features of the signal transduction process are conserved and which features are unique to specific receptors. We have previously explored the parathyroid hormone receptor-1 (PTHR1) and the glucagon-like peptide-1 receptor (GLP-1R), agonists of which are used to treat osteoporosis or type 2 diabetes, respectively. Continuing work on these two receptors will be complemented by new studies with the glucagon receptor (GCGR; glucagon is used by diabetes patients to reverse hypoglycemic shock) and the GLP-2 receptor (GLP-2R; agonists are used to treat short-bowel syndrome). Proposed studies will harness unique capabilities of our group to explore the consequences of varying agonist conformational propensities in terms of signaling outcomes and receptor binding. Results should be very impactful because agonist design efforts motivated by therapeutic goals usually focus on stabilizing a specific receptor-bound conformation of the agonist, which is fully α-helical for the GPCR agonists we study. Our recent findings raise questions about this approach; we have discovered surprisingly potent agonists based on designs that destabilize the α-helical conformation. The other major research focus is liquid-liquid phase separation (LLPS) mediated by proteins. LLPS in cells leads to a variety of “membraneless organelles” (MOs) that appear to be crucial for cellular function. The noncovalent interactions that underlie these dynamic assemblies are not well understood. Evolution of dynamic MOs toward more ordered assemblies appears to underlie some neurodegenerative diseases. Our approach involves developing models of protein-mediated LLPS in which one component is a peptide of < 50 residues and therefore readily accessible via chemical synthesis; the other component is a larger biopolymer (protein or RNA). This design strategy enables incisive evaluation of the roles of diverse noncovalent interactions in the dynamic associations that are unique to MOs. Results should inform future efforts to address diseases associated with “hardening” of MOs within cells.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT The overall goal of this revised proposal is to address the important unmet need for noninvasive detection and assessment of liver fibrogenesis, the pathologic process resulting in liver scarring and fibrosis. Fibrosis is the cumulative end-result of all chronic liver diseases (CLD) that affect ~1.5 billion people globally. This study will be enabled by using positron emission tomography (PET) through specific in-vivo targeting of the activated (fibrogenic) hepatic stellate cells (HSCs), the cornerstone of liver fibrogenesis and fibrosis. While non-invasive techniques for assessment of fibrosis have been developed and validated (e.g., magnetic resonance elastography, MRE), there is lack of a much-needed non-invasive biomarker to measure fibrogenesis, which is especially significant in the setting of drug-discovery. Development trials and eventual clinical im- plementation of antifibrotic drugs require non-invasive detection and quantification of active fibrogenesis. As a current limitation, assessment of treatment response through measuring fibrosis requires several months due to the time lag between reduction in fibrogenesis and subsequent decrease in fibrosis. Moreover, even if a drug is effective, MRE may not detect a change in liver stiffness despite improvement in histologic, serologic, and/or other imaging markers. Hence, non-invasive assessment of liver fibrogenesis is urgently needed. We have demonstrated successful non-invasive preclinical assessment of liver fibrogenesis through a fundamentally innovative strategy using PET. Fibroblast activation protein (FAP) is specifically expressed by activated HSCs. We have demonstrated that in our human-sized model of alcohol-induced liver injury, uptake of radiolabeled FAP inhibitor (FAPI) on PET directly correlates with fibrogenesis and fibrosis. Moreover, mouse studies have demonstrated reduced FAP expression with fibrosis reversal. However, there is need for sys- tematic evaluation of the relationship between FAPI PET and tissue markers of fibrogenesis (1) across all grades of liver injury, and (2) in both presence and absence of the hepatotoxin. This requires further preclinical evaluation of FAPI PET in a controlled setting and subsequent translational evaluation in patients. We aim to (1) Determine the relationship between liver FAPI uptake and tissue markers of fibrogenesis across all stages of liver injury in our swine model of liver injury; (2) demonstrate that decreasing liver FAPI uptake is a biomarker for decrease in fibrogenesis and its tissue markers, preceding any reversal in fibrosis; (3) determine the rela- tionship between liver FAPI uptake and tissue markers of fibrogenesis in a pilot clinical study in CLD patients. Successful completion of these aims by our unique and highly qualified multidisciplinary team will lay the foundation for future clinical trials for definitive validation in patients and to fulfill the unmet need of a non- invasive biomarker for liver fibrogenesis. The development of this biomarker will have a meaningful impact on the care of ~1.5 billion people with CLD through (1) facilitating the development of antifibrotic pharmacothera- peutics (2) early detection of fibrogenic liver disease, and (3) monitoring of fibrogenesis in response to treatment.

NIH Research Projects · FY 2026 · 2024-01

Abstract Chronic stress is pervasive in our society and linked to a reduced quality of life and increased risk for developing chronic diseases. Treatment options, including changes in lifestyle, exercise, and anxiolytics, are only partially effective and would benefit from a deeper understanding of the cellular mechanisms underlying the negative effects of chronic stress on the brain. Here we start by focusing on key restorative processes that occur during the inactive phase - the light period in rodents and the dark period in humans - when animals are asleep and for many hours brain activity is characterized by the presence of spindles and slow waves. These restorative processes involve integrated changes in astrocytes and neurons and include glymphatic clearance and renormalization (weakening) of synaptic strength. The former, by removing metabolic waste, reduces the risk of toxic waste build-up and the latter, by reducing the strength of synapses and the size of the perisynaptic astrocytic coverage, keeps the cost of synaptic activity under control and promotes future learning. It has been known for decades that the activity of the noradrenergic neurons of the locus coeruleus (LC) decreases during the inactive phase, leading to a progressive decline in noradrenaline (NE) levels in the brain. However, we and others have recently found that this decline is not monotonic: there are large, infraslow NE oscillations (INO) that occur in association with spindles and slow waves. We postulate that INO, combined with progressively lower NE levels and slow waves, are key to promote the cellular restorative processes occurring during the inactive phase. Based on this observation, we will use a mouse model of chronic stress to test the novel hypothesis that chronic stress disrupts the slow wave rich inactive phase and its complex NE dynamics, thereby impairing both astrocytic and neuronal restorative processes, including glymphatic clearance and renormalization of synaptic strength and of perisynaptic astrocytic coverage. To test our hypothesis we will combine recordings of brain activity (EEG/EMG), neuromodulators’ dynamics (voltammetry and NE sensors), serial block-face scanning electron microscopy, and brain fluid dynamics, along with optogenetic manipulation of NE/LC. We will use memory performance and sensitivity to painful stimulation as behavioral measures of the cellular restorative processes. Finally, we will test whether a non-pharmacological intervention, mild acoustic stimulation, can counteract the negative effects of chronic stress on astrocytes and neurons by restoring normal INO and slow waves. The proposal is organized in 2 aims. Aim 1 will define the cellular targets for the NE dynamics, including glymphatic clearance, synaptic weakening and renormalization of astrocytic synaptic coverage and glycogen storage. Aim 2 will test whether mild auditory stimulation in chronic stress can boost slow waves and normalize INO, thus improving the cellular restorative processes, memory performance and pain threshold. The proposed experiments are challenging but will benefit from the complementary expertise of the Nedergaard and Cirelli labs. If successful, the experiments will provide fundamental new insights into the biology of both astrocytes and neurons and their dysfunction in chronic stress.

NIH Research Projects · FY 2025 · 2024-01

Project Summary/Abstract Proper mitotic spindle positioning and orientation of the cell division plane is imperative for tissue morphogenesis, development, and determination of cell fate. When the spindle is mislocalized or the division plane is askew, defects impacting the genetic content of daughter cells, tissue architecture, and development of tumors may arise. In vertebrates, cytoplasmic dynein and its cortically localized regulators, such as Leu/Gly/Asn-repeat containing protein (LGN) anchor the spindle and provide the mechanical force needed to move the spindle into its proper position and orientation. The location of this spindle positioning machinery is, in turn, controlled by a variety of factors including polarity proteins, cell-cell junctions, or both, depending on the cellular context. In addition, studies going back more than a century have identified the so-called “default” spindle positioning mechanism, wherein the spindle spontaneously positions itself parallel to the longest axis of the cell and perpendicular to its shortest axis. In the absence of any other information, this default mode of spindle placement ensures formation of two equally-sized daughter cells. The small GTPase, Cdc42, has been repeatedly implicated as an upstream participant in spindle orientation in a variety of organisms and tissues, based largely on knockdown or knockout approaches. However, the means by which Cdc42 controls spindle positioning is poorly understood, in part because it can impact many processes known to be involved in spindle positioning including cell polarity, cell-cell junction formation, and cell-substrate adhesion formation. Further, almost nothing is known about Cdc42 dynamics during spindle positioning. Our preliminary data show that Cdc42 is present in propagating cortical waves during spindle positioning in Xenopus embryos, and that experimental suppression of these Cdc42 waves results in inappropriately positioned cleavage planes, a hallmark of mispositioned or misoriented spindles. I will use a combination of imaging, global and optogenetic manipulation of Cdc42, and knockdown approaches in intact Xenopus embryos and dissociated blastomeres to test the hypotheses that Cdc42 controls proper spindle positioning and to test whether it acts on the spindle positioning machinery directly or whether it acts through upstream control mechanisms such as cell polarization. In Aim 1, I will assess the role of Cdc42 in spindle positioning in intact embryos and dissociated blastomeres via both global and optogenetically controlled Cdc42 wave modulation. This will allow me to determine where and when in the spindle positioning hierarchy Cdc42 acts. In Aim 2, I will identify the Cdc42 activator, GEF, and inactivator, GAP, responsible for regulating Cdc42 waves. This will allow me to understand the molecular regulation of cortical Cdc42 dynamics during spindle positioning, and provide a complementary approach for wave manipulation. Collectively, these studies will reveal how Cdc42 controls proper spindle positioning in a vertebrate model.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY Transcription factors and cellular signaling mechanisms establish and maintain regulatory networks that allow the hematopoietic system to respond to infection, inflammation and other stresses. These networks mediate the expansion, mobilization, and differentiation of hematopoietic stem and progenitor cells (HSPCs). As exemplified by human germline variation in genes encoding HSPC-regulatory factors, such as GATA2 and RUNX1, genetic variation can disrupt HSPC activities, leading to recurring infection, cytopenia, and/or bone marrow failure. However, the mechanisms underlying these defective responses are incompletely understood. We have modeled human pathogenic variants causing GATA2 deficiency syndrome, where patients present with recurring infections and cytopenias with progression to MDS and AML. This model utilizes mice with a single-nucleotide variant from GATA2 deficiency patients, in conjunction with a severely impaired Gata2 +9.5 enhancer in the second allele (compound heterozygous; CH), mimicking epigenetic silencing and allele-specific expression correlating with disease presentation. We demonstrated that Gata2 variation alters steady-state HSPC levels, blocks HSPC expansion and differentiation in response to chemotherapy, attenuates long-term repopulating activity following bone marrow transplantation, leads to bone marrow failure following chronic inflammation by the viral mimetic polyI:C, alters HSPC response to the bacterial cell wall component LPS, and attenuates HSPC mobilization in response to pro-inflammatory cytokines, including G-CSF. We hypothesize that pathogenic clinical variants disrupt GATA2- and infection/inflammatory-instigated networks essential for HSPC expansion and differentiation, and we shall utilize global approaches to generate a rigorous foundation for elucidating how variants impact networks in the context of sterile- and infection-induced inflammation. Aim 1 will develop global insights into GATA2-dependent mechanisms that regulate HSPCs in response to bacterial infection and sterile inflammation. Barcoding cell surface markers coupled with single cell RNA-sequencing (CITE-seq), will be performed on HSPCs to determine pathways altered by the pathogen response, immunophenotypic populations affected, and how clinical variants disrupt the networks. The contribution of the niche to HSPC function will also be analyzed. Aim 2 will determine how fungal infection alters networks governing HSPC expansion, mobilization, and inflammation, and the requirement of GATA2 in these processes. We will generate libraries of differentially expressed transcripts in HSPC populations following infection with the pathogen Aspergillus fumigatus. Inflammatory cytokines will be quantified via multiplex cytokine analysis. We will test if networks responsible for HSPC response to fungal infection remain intact in GATA2 deficiency genetic models. These pilot studies will leverage existing models to generate systems and omic datasets to elucidate how germline genetic variation creates a predisposition to bone marrow failure.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY/ABSTRACT Developmental language disorder (DLD) impacts 7% of all children—both monolingual and bilingual— and has lifelong consequences for educational attainment and socioemotional wellbeing. DLD is characterized by deficits in structural language, but the mechanisms underlying these deficits remain under investigation. The Predictive Processing Hypothesis posits that prediction underlies human cognition and behavior, and it is increasingly being applied to language. Recent work has applied the Predictive Processing Hypothesis to DLD, investigating the possibility that deficits in predictive processing underlie language deficits. However, this hypothesis has only been tested in monolinguals, and only within the context of fluent, idealized language input. Fillers (e.g., uh and um), are a common disfluency in spontaneous language production and reliably precede difficult-to-retrieve words (e.g., words that are low-frequency). Neurotypical monolingual adults harness this association between fillers and production difficulty to anticipate upcoming novelty, but the mechanisms that underlie children’s comprehension of fillers as signals for upcoming difficult words are poorly understood. This is particularly so for children who have DLD and/or are exposed to more than one language (bilingual children). The overarching goal of this proposal is to examine both comprehension of fillers and consequences for learning in monolingual and bilingual children with and without DLD. A total of 80 children (40 English monolinguals and 40 Spanish-English bilinguals) ages 4-5, representing the full range of language ability (from DLD to typical language) will participate. The Specific Aims are: 1.) to determine the impact of disfluencies on language processing and learning in monolingual children with and without DLD and 2.) to determine the impact of disfluencies on language processing and learning in bilingual children with and without DLD. Two eye-tracking visual world tasks, in which children view novel and familiar objects side-by-side, will be used to assess predictive processing of fillers and word learning under fluent and disfluent conditions. Language ability (from typically developing to DLD) will be examined continuously under both aims, and bilingualism (proxied by years of second language exposure) will be examined continuously under Aim 2. Together, the findings across the two aims will test the innovative Predictive Processing Hypothesis within the novel context of disfluent speech and assess how consequences of disfluency on processing scale up to impact learning. Moreover, the findings will provide clinical insight into input characteristics that may enhance or disrupt learning and processing, with implications for structuring interventions. During the training period, the applicant will acquire new knowledge in the language profiles of bilingual and monolingual children with DLD as well as methodological considerations for working with this population; gain critical background knowledge in fluency and disorders; develop expertise in eye-tracking and advanced statistical analysis; and learn and practice principles of ethical science and effective mentorship.

NIH Research Projects · FY 2025 · 2024-01

Project Summary/Abstract Cell replacement therapies are one of the best options for multifactorial diseases of the retina like aged related macular degeneration (AMD), which normally cannot be treated by correcting mutations in individual genes. The retina is also highly surgically accessible, allowing stem cell derived neurons to be precisely transplanted and tracked post-operatively. While transplant therapies into the retina are ongoing, including Phase I human trials, success has been limited due to the inability of photoreceptors (PRs) to integrate properly into complex existing circuits. One potential means to improve PR integration is to transplant genetically modified PRs that have enhanced ability to extend processes and form synapses. We recently found that stem cell derived PRs have a limited time they are capable of autonomous axon extension, which correlated with a loss of an organized actin cytoskeleton within their terminals and an upregulation of synaptic proteins (Rempel et al., 2022). This result suggests that PRs become immobile due to their decreased ability to polymerize and organize actin, which is essential for cell and growth cone motility. Therefore, identifying age- dependent molecular changes that account for the loss of actin filaments and decreased PR terminal motility would provide potential targets to potentiate intrinsic PR terminal motility. Transplanting more motile older PRs that also express synaptic proteins could greatly improve circuit integration. It also should be noted that even young human PRs only extend axons an average of 20 um in isolation, so methods to improve terminal motility at any age could help restore circuits in the adult retina. Transplantation of “motility enhanced” PRs could greatly improve their integration into existing circuits. Here we propose a series of innovative experiments to first test likely upstream modulators that may be lost as PRs mature within retinal organoids (ROs). Next, we will conduct an unbiased proteomic screen of protein expression changes within PRs across relevant time points. Candidates will be tested to determine if PR actin dynamics, terminal motility and axon extension can be enhanced by increasing the activity of key proteins that show age-dependent loss of expression.

NIH Research Projects · FY 2026 · 2023-12

Natural killer cells provide a critical early defense against viral pathogens by virtue of their ability to recognize and kill virus-infected cells without prior antigenic stimulation. This is accomplished through the integration of signals from killer-cell immunoglobulin-like receptors (KIRs) and C-type lectin receptors (NKG2A and NKG2C) on NK cells and their MHC class I ligands on target cells. KIR and HLA class I polymorphisms can have profound effect on the course of HIV-1 infection and the efficacy of NK cell-based therapies for eliminating virus-infected cells. However, animal studies to address the underlying immunological mechanisms have been hampered by our limited understanding of NK cell receptor interactions in nonhuman primate models. For MERIT extension of R37 AI095098 “KIR and MHC class I immunogenetics in SIV infection”, we will build on recent success defining the MHC class I ligands of rhesus macaque KIRs, isolating antibodies to these receptors, and investigating the effects of viral peptides on KIR-MHC class I interactions. We will also extend these studies to assess the influence of viral peptides bound by MHC-E on interactions with macaque NKG2A and NKG2C and to test the hypothesis that adaptive NKG2C+ NK cells have a significant impact on the course of SIV infection. Aim 1 will determine how KIR polymorphisms shape ligand recognition and generate reagents (antibodies and MHC class I tetramers) for differentiating these receptors on primary NK cells. Aim 2 will investigate the influence of viral peptides bound by MHC class I ligands on KIR and NKG2A/C interactions to address alternative hypotheses concerning the role of peptides in enabling NK cells to differentiate virus-infected cells from healthy cells or to facilitate immune evasion. Aim 3 will assess the contribution of adaptive NKG2C+ NK cells to the outcome of SIV infection by depleting these cells during the acute and chronic phases of SIV infection. This animal study will also afford an opportunity to investigate longitudinal changes in the phenotypic properties of NKG2A+ and NKG2C+ NK cell subsets in response to SIV infection using KIR- specific reagents generated in Aim 1. RELEVANCE (See instructions): The proposed studies will provide an essential foundation for investigating natural killer (NK) cell responses to simian immunodeficiency virus (SIV) in rhesus macaques as a nonhuman primate model for HIV/AIDS and will explore fundamental mechanisms by which NK cells differentiate virus-infected cells from healthy cells and contribute to the control of immunodeficiency infection.

NIH Research Projects · FY 2025 · 2023-12

PROJECT SUMMARY/ABSTRACT As COVID-19 has illustrated, the emergence of novel RNA viruses from wild animals poses a significant threat to human health. In this proposal, we will begin to assess the zoonotic potential of an entire family of viruses: the arteriviruses. The arteriviruses are the only mammalian RNA virus family not known to infect humans; as such they have been largely ignored by the biomedical research community. However, these viruses cause severe disease in many other mammals including monkeys. Due to historical neglect, the research community has not developed the resources and knowledge that would be required to rapidly develop medical countermeasures in the event of an arterivirus emergence in humans. Thus, these viruses are uniquely positioned in our pandemic preparedness blindspot. In this project we will characterize the zoonotic risk posed by arteriviruses using a diverse collection of arteriviruses and variety of host tools. In Aim 1, we will build upon our recent discoveries that identified CD163 as a host molecule required for arterivirus entry into cells. We will create a panel of cells expressing CD163 orthologs from various species (human, monkey, mouse, pig) and examine the susceptibility of these cells to a panel of arteriviruses. Next, we will create CD163 molecules with mutations and deletions to map the key domains and residues required for arterivirus binding. Finally, we will use our knowledge of CD163 to try and grow novel arterivirus isolates that so far cannot be grown in culture. In Aim 2, we will build upon our recent discovery that the neonatal Fc receptor (FcRn) is required for arterivirus infection of cells. We hypothesize that FcRn acts as an attachment factor, allowing the virus to adhere to the cell surface and become internalized in the endosomal compartment where it contacts CD163. To test this, we will perform a series of binding and internalization assays (in the presence and absence of CD163) to establish a role of FcRn as a pan-arterivirus attachment factor. We will also overexpress CD163 and FcRn in the background of FcRn- and CD163-knockout cells to determine if these molecules are each required in sequential fashion or are redundant for arterivirus infection. Finally, we will perform a pair-wise species comparison of FcRn orthologs and divergent arteriviruses to examine whether FcRn acts as a barrier to cross-species infection. Upon conclusion of this project we will have a greater appreciation for the zoonotic risk posed by these viruses. We will also have developed tools, biological systems, and knowledge that will serve as a critical resource for the rapid development of life-saving medical countermeasures in the event of an arterivirus outbreak in humans.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY Hearing loss is one of the most common age-related diseases. Untreated hearing loss is associated with in- creased risks of negative social-emotional health outcomes including loneliness and depression. Why hearing loss is associated with poor social-emotional health is unknown. The primary treatment for hearing loss is hear- ing aids, but only 30% of adults with hearing loss use hearing aids. Whether hearing aids reduce the risk of poor social-emotional health is equivocal, and if they do moderate the risk, how they do is unknown. In order to improve the effectiveness of audiologic interventions in reducing the risk of poor social-emotional health, it is crit- ical to understand why hearing loss is associated with poor social-emotional health, whether the mechanisms of this association are hearing-related, and how hearing aid use moderates the risk of poor social-emotional health outcomes. This study addresses these questions by introducing a theoretical framework of hearing-related be- havior and novel methods to quantify it in the real world. Hearing-related behavior is proposed to comprise both auditory lifestyle, or how acoustically demanding and diverse the soundscapes a listener encounters in daily life are, and communication engagement, or how well a listener is able to engage in conversation within sound- scapes in daily life. In the proposed study, these domains are measured using long-form audio recordings and ecological momentary assessments collected from the daily lives of listeners with hearing loss who do and do not use hearing aids. Then, the relationship between these domains and social-emotional health is determined. In Aim 1, auditory lifestyle differences between the two groups is quantified by measuring auditory lifestyle demand (proportions of different types of soundscapes) and diversity (predictability of soundscapes) for each participant. Long-form audio recordings of daily life are analyzed using machine learning classification to precisely character- ize the soundscapes of each listener. Sound recordings are also paired with ecological momentary assessment to add context. Whether hearing aid users spend more time in demanding (e.g., speech-in-noise, group conversa- tion, unfamiliar talkers) and diverse (less predictable) soundscapes than non-hearing aid users is determined. In Aim 2, recordings from the real world are identified where participants engaged in conversation. Conversations are analyzed for utterance length and conversational turn rate, two measures of communication engagement. Differences in engagement between hearing aid users and non-hearing aid users are determined. In Aim 3, participants complete standardized and momentary measures of social-emotional health (depression, loneliness, quality of life). The association between hearing-related behavior, hearing aid use, and social-emotional health is determined. Findings from this study will determine how social-emotional health and hearing loss are related and how clinical audiology can best promote positive behavior change that reduces the risks of loneliness and de- pression in people with hearing loss. This study addresses goals of the NIDCD Strategic Plan by using machine learning to develop new outcome measures that will improve clinical practice and intervention effectiveness.

NIH Research Projects · FY 2026 · 2023-12

1. PROJECT SUMMARY/ABSTRACT Candidate: I am presently a scientist in the BRAVE Research Center (BRC), within the Department of Psychiatry at the University of Wisconsin (UW). Career Goals: I seek to identify neural mechanisms though which childhood adversity can perturb typical development of biobehavioral affective systems in ways that effect enduring psychiatric risk, as well as moderative ecological factors that may provide a foundation for evidence-based clinical efforts with vulnerable youth. Career Development: I request support for mentored training to build knowledge and skills in four areas pertinent to my career goals: (1) theory and analysis of functional brain networks; (2) caregivers and caregiving as ecological moderators of the linkage between victimization and psychopathology; (3) data-driven approaches to neurobehavioral analyses and (4) professional development as an independent investigator. Research Project: The proposed work utilizes longitudinal, multimodal assessment data from parent-child dyads participating in the Adolescent Brain Cognitive Development study (N=11,875), and its Social Development sub-study (N=2,500). The primary aim is the evaluation of cross- sectional and longitudinal changes in functional connectivity networks as mechanisms linking childhood adversity (victimization) and caregiving practices to impairments in emotion regulation, a biobehavioral domain with transdiagnostic relevance. First, I will extract robust functional network (static and dynamic) data from quality- controlled ABCD resting state fMRI scans and evaluate these metrics (and their change across time) as mediators in a structural equation model linking victimization and emotion regulation. Second, I will consider main effects of caregiving practices (e.g., positive parenting) on functional network development, as well as modulative impact on neurofunctional sequelae of victimization. Third, I will develop and evaluate a deep- learning model predicting impairment in emotion regulation from neural and behavioral inputs, including regional connectivity, caregiving practices, and exposure to victimization. These results will inform future applications proposing independent research projects considering neurofunctional mechanisms linking adversity to psychiatric risk in the context of granular assessment of caregiving practices, caregiver-child relationships, and dynamics within the broader family system. Such findings may serve as the basis for developing targeted clinical approaches translating ecological factors shown to moderate the biological and behavioral sequelae of victimization. Environment: Activities in this proposal will be conducted at UW (BRC, Department of Psychiatry) under the mentorship of esteemed scientists working in areas of affective and developmental neuroscience, neural and behavioral sequelae of adversity, quantitative modeling of developmental processes, and machine learning, led by Drs. Ryan Herringa, Seth Pollak, and Vikas Singh (respectively), each of whom possesses extensive experience mentoring early career investigators.

NIH Research Projects · FY 2025 · 2023-12

PROJECT SUMMARY Diabetes mellitus has been characterized since antiquity, with the first documentation of this group of disorders dating back to 1500 BC1. While many advances have been made in the field in terms of treatment, there remains no cure for diabetes. This has become increasingly problematic for both the individuals and the entire healthcare system as rates of diabetes continue to rise. It is estimated that healthcare for individuals with diabetes costs over $200 billion per year in the United States alone2. Diminishing these costs and improving treatment options for patients are just a couple of the many reasons why diabetes-related research is so critical. I will use a genetic approach to more fully understand the initiation and progression of diabetes. In this application, I highlight a method that integrates human and mouse datasets to examine the role of intragenic and intronic regions of the genome in diabetes susceptibility. One such locus of interest resides in the first intron of the GLIS3 gene and has highly significant SNPs associated with both type 1 diabetes (T1D) and type 2 diabetes (T2D). To study this Glis3T1D/T2D locus, we determined the syntenic region in mouse and deleted 1729bp to create a novel mouse model (Glis31729 mouse). I found that these mice exhibit hyperglycemia, despite increased insulin secretion and glucose-evoked Ca2+ oscillations. My overall objective for this proposal is to functionalize this locus in an effort to elucidate its role in driving diabetes. My central hypothesis for the function of this locus is that it acts as an enhancer for Glis3 in islets, and that T1D/T2D-related SNPs within the locus alter its activity. I will test this hypothesis, and thus accomplish the goals of this proposal, by pursuing the following aims: 1) evaluate the enhancer activity of the Glis3T1D/T2D locus, 2) identify the influence of T1D/T2D-associated SNPs on transcription factor binding at the Glis3T1D/T2D locus, and 3) evaluate differentially expressed (DE) genes in the Glis3 1729 mice to reveal the role of the Glis3T1D/T2D locus on altering T1D and T2D risk. With the completion of these aims, I anticipate a more complete understanding of this non-coding locus, including how it regulates gene expression and affects both type 1 and type 2 diabetes susceptibility. Additionally, I believe this proposal will provide the necessary training for me to progress as an islet biologist and prepare me for a tenure-track position at a research institution.

NIH Research Projects · FY 2026 · 2023-12

Project Summary/Abstract Background – The microbiome affects host metabolism predominantly via metabolites synthesized or modified by gut bacteria. The identity of these metabolites and their mechanisms of action in the host remain largely unknown. Overarching metabolomic analyses have offered a glimpse into classes of microbial molecules and how they associate with disease. Two such types of molecules, bile acids and folates have been shown in recent years to differentially modulate cell signaling pathways. However, the mechanisms of how distinct bile acids and folates induce gut permeability and inflammation, the hallmarks of metabolic syndrome remain largely evasive. Research – Targeted metabolomic analyses in human patient and rodent models of inflammatory diseases will identify bile acid and folate metabolites that differ compared to healthy controls. Preliminary data suggests that microbial unconjugated bile acids induce intestinal permeability, inhibition of which protects against development of Non-Alcoholic Steatohepatitis (NASH). Bile acids are deconjugated by gut bacteria. The K99 phase of the proposal will identify mechanisms of how unconjugated bile acids induce intestinal permeability, and if chronic inhibition of gut bacterial bile acid deconjugation is an effective strategy to rescue gut permeability and NASH. There is sufficient evidence to suggest that bacterially produced polyglutamylated (glu) folates can activate host folate receptor (FR) signaling, which in turn induces the MAP kinase pathway. The K99/R00 transition phase of the proposal will identify and quantify individual folate molecules in diseased cohorts that activate FR. The diseased samples include rodent and human bariatric surgery, NASH, and Inflammatory Bowel Disease (IBD) intestinal contents. A compound library of folates will be generated for high-throughput screening. Inflammatory folate producing bacteria will be isolated to establish causal relationships between strains and gut inflammation. Multiple IBD patient ileal biopsies show an upregulation of the folate hydrolase (FOLH1) gene, the only known purpose of which is to deconjugate poly-glu folates to mono-glu form. In the R00 phase, the mechanism of folate- mediated upregulation of FOLH1 in IBD will be studied. Further, the role of poly-glu folate deconjugation will be studied in the context of intestinal inflammation and metabolism. Alterations in mitochondrial dynamics, one- carbon metabolism, and energy status will be measured following activation or inhibition of the folate/FR/FOLH1 axis. This study will also utilize high-throughput screening to identify molecules that can rescue gut inflammation. Impact on Public Health – Bacteria in the gut encounter bile and dietary nutrients prior to their absorption in the body. Approximately 50% of bile acids and folates absorbed from the intestine is microbially derived. Therefore, studying microbiome-derived metabolites and their activity is important not only for intestinal, but organismal homeostasis. Bile acids are one of the most abundant molecules in the gut, present in millimolar concentrations. Folate concentrations, also high in the gut, vary widely based on folate fortification policies imposed by different countries. Therefore, the study of these metabolites in disease prognosis will reveal strategies for amelioration.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY 18F-Fluorodeoxyglucose (FDG) PET/CT imaging has become an essential tool for guiding and adapting treatments for lymphoma. However, the PET evaluation criteria currently used for assessing lymphoma, which consists of subjective visual scoring on a 5-point scale, is suboptimal. The visual scores suffer from high inter- observer variability and have low prognostic power for new emerging biological therapies. Quantitative PET metrics have been shown to be more predictive of clinical outcomes than visual scores, but quantitative analysis of whole-body PET/CT images is prohibitively time-consuming and impractical in routine clinical care. Deep learning (DL) has shown promise in automating the quantitative analysis of baseline FDG PET/CT images, but comprehensive evaluation of interim-therapy and post-therapy images using DL has proven difficult. Residual lymphoma has low-level uptake, which can be hard to differentiate from physiologic or treatment-related uptake, and reading physicians must use clinical histories and baseline PET images (i.e., sites of initial disease) to make reliable diagnoses. DL algorithms, on the other hand, only operate on cross-sectional images and are unable to account for historical context. Our objective is to develop DL algorithms that operate on PET/CT images from more than one time point so that algorithms can learn longitudinal dependencies for contextually-aware predictions. We also aim to develop multimodal vision-language models that can simultaneously interpret radiology text reports while performing PET/CT image analysis. These models can leverage critical information about patient history and physician interpretation when processing retrospective images. Furthermore, we will use semi-supervised learning to leverage both unlabeled datasets and labeled datasets. Our overall goal is to develop contextually-aware algorithms for automated longitudinal analysis of whole-body PET/CT images in lymphoma. These tools will be developed using diverse datasets from multiple institutions. PET metrics measured by DL will be validated as predictive markers of outcome using data from a Phase 3 clinical trial.

NIH Research Projects · FY 2024 · 2023-09

Responding to NIH’s Helping to End Addiction Long-term (HEAL) initiative NOSI titled “Opioid Use Disorder Care Pathways for Individuals with Histories of Exposure to Violence,” this R61/R33 project seeks to develop and evaluate a brief video and text messaging intervention delivered in the emergency department (ED) to prevent the onset or escalation of opioid use, misuse, or disorder among recent sexual assault (SA) survivors. More than 1 in 4 women will experience an attempted or completed rape in her lifetime. PTSD and opioid misuse are prevalent in the wake of sexual violence, and the opioid epidemic is having a unique impact on women, with sharper increases in prescription opioid deaths, a greater likelihood of meeting criteria for opioid use disorder (OUD), and increased likelihood of comorbid mental health conditions compared to men. ED visits for sexual assault medical forensic exams (SAMFEs) have increased more than 1500% in the last two decades and may provide an important opportunity to intervene to prevent the onset or escalation of PTSD and opioid misuse and disorder. Our team developed and tested a brief video intervention delivered in the ED to women presenting for a SAMFE in two randomized controlled trials. Compared to treatment as usual (TAU), the video was associated with reductions in PTSD, depression, suicidality, alcohol, marijuana, and nicotine use for subgroups of survivors when followed over 6 months. However, in our second RCT, an active control mindfulness video better reduced post-SA opioid use compared to TAU for survivors with pre-SA opioid misuse. Thus, we propose to develop a new video, Skills Training in Active Recovery (STAR) based on the evidence-based principles of both videos to better address PTSD and opioid misuse. In the previous RCTs, survivors reported substantial distress at the SAMFE; given that distress can impair memory encoding, we propose to develop a 3-week daily text messaging program called TextSTAR that will reinforce content presented in the video but give opportunities for survivors to engage with the material in a different setting and over a longer period of time. During the R61, we propose to develop the STAR video and TextSTAR and get feedback from a community advisory board of sexual assault survivors about the wording and presentation of the content to maximize acceptability and utility. We will pilot test STAR and TextSTAR with a separate sample of survivors (N = 50) recruited from 5 ED sites within the Better Tomorrow Network, a research network of SAMFE programs throughout the US, to gather preliminary data about our recruitment approach, acceptability of the intervention, and efficacy. In the R33, we will conduct a SMART trial where survivors will be randomized to receive STAR or no video at the ED, assessed at 1 week, and those above threshold for acute stress/PTSD or opioid use, misuse or disorder will be randomized to receive TextSTAR or no text intervention for 3 weeks. We will re-assess PTSD and opioid misuse and disorder at 1, 3, and 6 months. If efficacious, this intervention is a low-cost and easy-to-disseminate approach to reduce PTSD and opioid outcomes among SA survivors.

NIH Research Projects · FY 2025 · 2023-09

Project Summary Increased oxidative stress is associated with cardiac cell dysfunction in heart disease. An unbalanced redox state leads to an increase in the post-translational modification of S-Glutathione, which modifies cysteine residues on key myofilament proteins, such as cardiac myosin binding protein-C (cMyBP-C). cMyBP-C regulates contraction and relaxation of the sarcomere. The phosphorylation of cMyBP-C by Protein Kinase A (PKA) is cardioprotective. Yet, in the failing heart, phosphorylation levels of cMyBP-C are reduced, contrary to an increase in S-glutathionylated cMyBP-C. When cardiomyocytes were incubated with oxidized glutathione (GSSG), myofilament calcium sensitivity increased and cross-bridge kinetics slowed. Prior experiments were unable to isolate the specific effects of S-glutathionylated cMyBP-C (i.e., without the effects of other S- glutathionylated proteins) nor the specific sites responsible for the functional change. Phosphorylation and S- glutathionylation of cMyBP-C may have antagonistic effects. Incubating three N’-terminal domains of cMyBP-C with GSSG led to a significant increase in S-glutathionylated cMyBP-C and a downregulation in cMyBP-C phosphorylation. These results indicate that the decrease in cMyBP-C phosphorylation and consequent loss of the cardioprotective effect of phosphorylated cMyBP-C seen in the failing heart could be due to an increase in S-glutathionylated cMyBP-C. In addition, the anti-ischemic drug, ranolazine, has been shown to improve diastolic function to sham levels, which correlated with S-glutathionylated cMyBP-C. These data indicate that a currently available cardiac therapy might be useful in moderating the levels of cMyBP-C S-glutathionylation. Thus, this proposal will identify how the interaction between phosphorylation and S-glutathionylation affects cardiomyocyte function under normal, elevated, and therapeutically treated conditions using our novel “cut and paste” SpyC3 mouse model. Protein domains C0C7 of cMyBP-C will be “cut” from the sarcomere using the tobacco etch viral protease and, after washing steps, recombinant C0C7sc protein with and without modified cysteine residues will replace its location within its endogenous location in the sarcomere. Site-directed mutagenesis will be used to generate cysteine substitution constructs preventing S-glutathionylation at specific residues and the functional effects of each construct will be measured using the “cut and paste” model. Dual incubation of PKA and GSSG with and without ranolazine treatment will determine the functional effects of this interaction. A combination of ProQ Diamond staining, immunoblotting, Phos-tag gels, and mass spectrometry will be used to measure total modification levels and identify the site-specific modifications. Results from this proposal will be the first to identify the functional effects of individual cMyBP-C S-glutathionylated residues and of phosphorylation and S-glutathionylation cMyBP-C crosstalk. This research proposal will lead to a better understanding of the effects of oxidative stress on cMyBP-C function, its potential to affect phosphorylation in the heart, and if a currently available therapeutic might benefit hearts affected by oxidative stress.

NIH Research Projects · FY 2026 · 2023-09

Responding to NIH’s Helping to End Addiction Long-term (HEAL) initiative NOSI titled “Opioid Use Disorder Care Pathways for Individuals with Histories of Exposure to Violence,” this R61/R33 project seeks to develop and evaluate a brief video and text messaging intervention delivered in the emergency department (ED) to prevent the onset or escalation of opioid use, misuse, or disorder among recent sexual assault (SA) survivors. More than 1 in 4 women will experience an attempted or completed rape in her lifetime. PTSD and opioid misuse are prevalent in the wake of sexual violence, and the opioid epidemic is having a unique impact on women, with sharper increases in prescription opioid deaths, a greater likelihood of meeting criteria for opioid use disorder (OUD), and increased likelihood of comorbid mental health conditions compared to men. ED visits for sexual assault medical forensic exams (SAMFEs) have increased more than 1500% in the last two decades and may provide an important opportunity to intervene to prevent the onset or escalation of PTSD and opioid misuse and disorder. Our team developed and tested a brief video intervention delivered in the ED to women presenting for a SAMFE in two randomized controlled trials. Compared to treatment as usual (TAU), the video was associated with reductions in PTSD, depression, suicidality, alcohol, marijuana, and nicotine use for subgroups of survivors when followed over 6 months. However, in our second RCT, an active control mindfulness video better reduced post-SA opioid use compared to TAU for survivors with pre-SA opioid misuse. Thus, we propose to develop a new video, Skills Training in Active Recovery (STAR) based on the evidence-based principles of both videos to better address PTSD and opioid misuse. In the previous RCTs, survivors reported substantial distress at the SAMFE; given that distress can impair memory encoding, we propose to develop a 3-week daily text messaging program called TextSTAR that will reinforce content presented in the video but give opportunities for survivors to engage with the material in a different setting and over a longer period of time. During the R61, we propose to develop the STAR video and TextSTAR and get feedback from a community advisory board of sexual assault survivors about the wording and presentation of the content to maximize acceptability and utility. We will pilot test STAR and TextSTAR with a separate sample of survivors (N = 50) recruited from 5 ED sites within the Better Tomorrow Network, a research network of SAMFE programs throughout the US, to gather preliminary data about our recruitment approach, acceptability of the intervention, and efficacy. In the R33, we will conduct a SMART trial where survivors will be randomized to receive STAR or no video at the ED, assessed at 1 week, and those above threshold for acute stress/PTSD or opioid use, misuse or disorder will be randomized to receive TextSTAR or no text intervention for 3 weeks. We will re-assess PTSD and opioid misuse and disorder at 1, 3, and 6 months. If efficacious, this intervention is a low-cost and easy-to-disseminate approach to reduce PTSD and opioid outcomes among SA survivors.

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract The increase of people with complex chronic health conditions is stressing the U.S. healthcare delivery system. People needing chronic care management (CCM) are at a high risk of medication safety issues and health- related harms, such as hospitalizations and emergency department visits. Community pharmacies play a role in ensuring patients’ safe medication use for CCM, but their efforts are undermined by volatile work demands and other system barriers. In preventing medication hazards for people needing CCM, pharmacy staff are further hampered by the historical preoccupation with what “went wrong” (a Safety-I approach), rather than also considering the vast majority of times that things “go right” (a Safety-II approach). This project seeks to reengineer the pharmacy system to advance a combined Safety-I and Safety-II approach to improve CCM. Our transdisciplinary team of pharmacists, health services researchers, engineers, and quantitative and qualitative researchers will partner with pharmacies in 2 large healthcare organizations (Advocate Health, UW Health), and Boscobel and Center independent pharmacies, to design and evaluate a pharmacy work system of care that optimizes resilience. To achieve this objective, we will create the Medication Safety Map (MedSafeMap) for pharmacists and technicians to better navigate complex pharmacy tasks, and to facilitate communication with patients and clinicians, while safely providing medications for patients dealing with CCM. The CARE (Concepts for Applying Resilience Engineering) and SEIPS (Systems Engineering Initiative for Patient Safety) models will be used to define and address the complex pharmacy work system issues that exacerbate medication safety risks for patients needing CCM. Human factors and systems engineering will be used to carry out the AHRQ RFA-required 5-step method. Aim 1 will focus on problem analysis of the work system through pharmacist and technician stakeholder observations and interviews, and analyses of work system information flow and artifacts, which will illustrate relationships among work system features that are then addressed in Aims 2 and 3. Aim 2 will employ participatory design and in-situ simulations to develop and design MedSafeMap detailing pharmacy redesign recommendations to optimize pharmacist and technician interactions within their work system and with patients/caregivers. For Aim 3, we will implement and evaluate MedSafeMap impact on CARE/SEIPS resilience outcomes – pharmacy staff attitudes, behaviors, performance, and work demands (interviews and surveys), time involved in tasks (time and motion analysis), types and quantities of services provided, and perceived changes in medication-related problems (self-reported data). This innovative transdisciplinary PSLL seeks to redesign interactions to strengthen pharmacy staff resilience while ultimately improving medication safety for patients needing CCM. This study, therefore, responds to an AHRQ priority population (i.e., patients with chronic disease) and a patient safety priority “to examine effective ways to make system-level changes to help prevent errors” that span the medication use process continuum.

NIH Research Projects · FY 2025 · 2023-09

Most microorganisms live in communities containing hundreds or thousands of species, each engaging in a rich web of interactions. The complexity of these interactions makes quantitative predictions about community dynamics difficult. To overcome this, simple proxies for natural communities, designated "model microbial communities," have been designed to support laboratory study. These models are complex enough to exhibit community-specific phenomena but simple enough to reveal governing principles of community interaction. Invasions are among the most destabilizing events that a microbial community can experience, often resulting in community dysfunction or host disease. We propose to use THOR, a model community that we developed, to characterize the response to invasion by Pseudomonas aeruginosa. We will track population dynamics functionally profile molecular interactions. The multi-omics and multifactorial nature of this study present multifaceted opportunities for statistical innovation. A truly integrative analysis cannot simply perform parallel hypothesis tests across assays, and there is a need for a differential testing framework that blends data sources into a unified molecular interaction network. We will draw from advances in selective inference and multi-omics network analysis to develop methods that illuminate the molecular interactions driving community response. This will allow us to tailor interventions that shape dynamics in the THOR model microbial community. We propose: 1. Aim 1: Functionally profile THOR's community response to P. aeruginosa using metabolomics, metatranscriptomics, and 16S rRNA sequencing and establish associated data curation workflows. These are the core data-generating experiments and quality control steps that provide accurate and complementary views of THOR under invasion. 2. Aim 2: Develop differential testing methods that are sensitive to interaction effects and that control module-level false discovery rates. We will introduce methods for selective inference of differential interactome modules, like activated biosynthetic pathways. 3. Aim 3: Consolidate software for differential interactome analysis and experimentally validate knockout targets. These experiments will illustrate a data-driven approach to control invasion dynamics, and our software will make such analysis easily accessible.

NIH Research Projects · FY 2025 · 2023-09

Project Summary Recent breakthroughs in neuromodulation for diet and weight control have stimulated a growing interest in the development of new anti-obesity strategies. However, achieving effective, real-time, and maintenance-free electrical neuromodulation with minimal side effects remains a major challenge. To address this challenge, this project proposes to develop a battery-free, flexible, and implantable piezoelectric nanogenerator (NG) that produces closed-loop, biofeedback electrostimulation (ES) on the vagus nerves to control food intake in response to stomach motions. This project builds on the collaborative work by Wang (PI) and Cai (co-I) of an implantable vagus nerve stimulation (VNS) device, which achieved effective diet and weight control in rats. The battery- and electronics-free VNS device is attached to the stomach surface and generates alternative current (AC) ES signals to the vagus nerves only when the stomach moves upon food intake. Our preliminary study demonstrated 38% less weight gain on normal adult rats with the VNS device implantation as compared to controls over a 100-day testing period. Although this efficacy value surpassed most peer reports, the ES signal intensity was 1-2 orders of magnitude smaller compared to those typically used. We hypothesize that tuning the closed-loop ES signal to the typical level of neuromodulation may further increase weight loss efficacy outperforming the currently-used non-natural continuous ES. To test this hypothesis and eventually bring this intriguing technology to clinic, we propose to develop a piezoelectric NG that provides tunable ES pulse signals up to 10 V in response to stomach peristalsis, and remains safe and stable over long- term implantation. We will also optimize the implantation of the VNS device and validate the closed-loop VNS efficacy and advantages to using standard obese rat models. In Specific Aim 1, we will develop a biomaterial- based flexible piezoelectric NGs that can produce tunable ES pulses in response to simulated stomach movements. In Specific Aim 2, we will evaluate the biocompatibility of the NG ex vivo and in vivo on the stomach of rats, and examine implantation sites and in vivo outputs in correlation to stomach motions. In Specific Aim 3, we will quantify and compare the diet and weight control performances on two obese rat models among three different strategies of using on-stomach NGs for VNS: (1) battery-powered open-loop VNS; (2) NG-enabled self- powered closed-loop VNS; (3) NG-switched battery-powered closed-loop VNS. This project will deliver a novel biomaterial-based VNS device that is battery- and electronics-free for weight control. This project uses rat model to test the new VNS devices, providing rapid feedback for device optimization, and quantifying the therapeutic efficacy in correlation to ES signals. Implantation-related technical issues will also be addressed. Together, we will establish an essential biological and engineering foundation that will allow us to move rapidly to the next step of in vivo studies in large animal models, eventually leading to human trials.