University Of Wisconsin-Madison

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY The outcome for patients with HER2-positive breast cancer (BC) has been significantly improved with the use of HER2-directed agents; however, hormone receptor (HR)+/HER2+ BCs that express both HR and HER2 are less responsive to anti-HER2 therapies as compared with HR-/HER2+ BC. Combined endocrine therapy plus anti-HER2-targeted therapy have been clinically investigated, but only a small subset of patients benefit from the treatment, indicating that HR+/HER2+ tumors have different biological characteristics, such as ER and HER2 signaling crosstalk. Dual HER2-blockade induces a Luminal A-like phenotype both in patients’ tumors and in in vitro models. However, the molecular mechanism underlying activation of ER signaling by anti-HER2 agents remains unclear. Using single cell RNA-sequencing (scRNA-seq), we identified Bromodomain Containing Protein 8 (BRD8) as an essential hub bridging HER2 and ER signaling pathways. We found that, BRD8, a component of p400 histone acetylase complex, was rapidly induced by treatment with a variety of anti-HER2 agents including neratinib, lapatinib and trastuzumab and that BRD8 is at the high hierarchy of ER activation in responsive to HER2 blockade. Furthermore, BRD8 regulates other growth promoting pathways in addition to ER. BRD8 knockout significantly impairs growth of a fulvestrant-resistant cell line and xenografts, and BRD8 knockdown enhances the sensitivity of HR+/HER2+ cells to HER2-targeting agents. These preliminary data lead to the hypothesis that BRD8 is an essential hub linking ER and HER2 pathways and BRD8 mediates ER activation in response to HER2 blockade in HER2+ cells. Therefore, combinatory BRD8 ablation with HER2 blockade should be effective for treating HR+/HER2+ BC. We propose three aims to test our hypothesis: (1) to elucidate the mechanism of BRD8 activation by anti-HER2 agents in HER2+ cells at the single cell level; (2) to define the roles of BRD8 in ER signaling activation and ER-independent functions upon HER2 blockade; (3) to test whether ablation of BRD8 sensitizes HR+/HER2+ BC to anti-HER2 blockade. The studies will reveal, at single-cell level, the cell-cell variability of BRD8 induction by anti-HER2 agents, and the novel functions of BRD8 in mediating ER/HER2 signaling crosstalk and ER-independent growth promoting functions. The mechanistic insights will lay foundation for developing combinatory BRD8 ablation and HER2 blockade therapy as a new treatment regimen for HR+/HER2+ BC.

NIH Research Projects · FY 2025 · 2023-03

ABSTRACT In the proposed work, I will be investigating how neural stem cells establish ‘proliferative asymmetries’, which are responsible for precisely controlling the development and repair of the central nervous system. During development and tissue repair, stem cells are triggered to proliferate. While proliferation is necessary, it must be precise to prevent overgrowth that can lead to tumorigenesis. As neural stem cells proliferate, they commonly divide asymmetrically. The ability to undergo asymmetric cell division is a highly conserved feature of neural stem cells across all animals. Asymmetric cell division produces one daughter cell that retains the neural stem cell identity, and one daughter cell which takes on a neuron-producing progenitor cell identity. A key difference between these two daughter cells is that the stem cell will rapidly reenter the cell cycle and divide again, while the progenitor cell will divide much more slowly or stop dividing all together. This proliferative asymmetry ensures that the proper number of neurons are produced, and its disruption leads to defects in neurogenesis. The molecular basis for this proliferative asymmetry mostly unknown. I will focus my independent research program around the mechanisms which establish this proliferative asymmetry. I hypothesize that this proliferative asymmetry is established by the differential inheritance of proliferation promoting factors during asymmetric cell division. I plan to learn how neural stem cells generate proliferative asymmetries in both invertebrates and vertebrates, through the use of Drosophila and zebrafish animal models. Studying both animal models will allow me to identify conserved and divergent modes of generating proliferative asymmetries in neural stem cells, while also providing me with new training opportunities. Both animal models are highly amenable to live cell imaging of neural stem cells in developing brains. In Aim 1, I will screening for proliferation regulators which are polarized in the mother neural stem cell. In Aim 2, I will determine how polarized proliferative regulators contribute to proliferative asymmetries between sibling cells. In Aim 3, I will determine how PAR polarity proteins regulate the PI3K proliferation pathway. Through my initial screening, I have already discovered one promising candidate that appears to mediate the proliferative asymmetry in the neural stem cell lineage, the lipid PIP3. PIP3 is transiently produced by the mother neural stem cell just before division, becomes polarized to one side of the cell, and gets inherited by the daughter cell which retains the neural stem cell identity. Discovering how PIP3 and other factors establish proliferative asymmetries during asymmetric cell division will advance our understanding of how neural stem cells mediate development and tissue repair. Through training activities aimed at improving my skills in grant writing, scientific teaching, and mentoring, I will be better prepared to run my own laboratory. The scientific discoveries and training this proposal will facilitate, will be foundational to building my independent research program.

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY Metabolic diseases are increasing worldwide in prevalence and severity. Globally, approximately 650 million people are obese or overweight, over half of whom are children. The healthcare burden of obesity is enormous and only growing. Circadian rhythms play a central role in healthy metabolism and in maintaining proper host- microbe interactions. Circadian rhythms in feeding and behavior create a feedback loop, driving rhythms in hepatic transcription, which in turn drive rhythms in host and microbial metabolism that ultimately feedback onto circadian rhythms in host behavior. This network of rhythms is severely disrupted in obese mice consuming a Western diet. Early-life diet interacts with the developing microbiome to program metabolism and disease risk, but how the early-life microbiota contribute to development of the host metabolic circadian network remains unknown. There is an urgent need to identify mechanisms by which gut microbes interact with diet early in life to shape these circadian networks that coordinate daily rhythms in metabolism. Mice devoid of all microbes (‘germ-free’) exhibit disrupted circadian rhythms in numerous aspects of host metabolism compared to mice that have had microbes since birth. Remarkably, repopulating germ-free mice later in adulthood with microbes (‘conventionalization') only partially normalizes their circadian rhythms. This indicates that the age at which gut microbes are acquired must play a critical role in the development of circadian networks that govern metabolism. But when and how healthy and obesogenic microbes first begin to normalize and disrupt, respectively, the circadian network remain unexamined. Our research will bridge this gap in knowledge by testing the hypothesis that the organismal circadian network is shaped by interactions between the type of gut microbes acquired and the developmental age of acquisition. Aim 1 tests the hypothesis that the age when mice acquire a normal, healthy microbiome determines the robustness of their circadian network in adulthood. We will conventionalize germ-free mice with microbes during gestation, at weaning, or in adulthood. Phenotypes across the entire circadian network will be examined later in life, including: hepatic clock and clock-controlled gene expression via RNA-seq, microbial community oscillations via 16S rRNA, and host behavioral circadian rhythms thermoregulation and via wireless telemetry inside sterile isolators. Aim 2 will test the hypothesis that obesogenic diets early in life disrupt normal development of the circadian network through a combination of microbe-dependent and microbe-independent mechanisms. Beginning early in life, germ-free mice will be administered an obesogenic diet alone, obesogenic microbes alone, or both obesogenic diet and microbes. Metabolic circadian networks will be examined as in Aim 1. These studies will specify how diet and microbes interact to affect the developing host-microbial circadian network. New insights into how metabolic programming impacts the circadian system will inform early-life dietary and microbial interventions that may afford prophylaxis against the development of obesity.

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY Liquid biopsy modalities that can non-invasively detect disease-associated biomarkers from biofluids can enable early cancer detection and patient monitoring with implications for improved survival rates. However, current methods have not achieved critical sensitivity and accuracy to be approved for population screening programs. New spectrochemical liquid biopsy methods, such as Raman and infrared spectroscopy, coupled with machine learning models are emerging as next-generation diagnostic modalities. Yet, fundamental physical limitations of light-matter interactions using conventional optical setups hinder the analytical performance of molecular spectroscopy techniques. Here, we propose to employ novel electromagnetic metasurfaces that can advance the analytical sensitivity and chemical selectivity of infrared absorption spectroscopy enabling its real-world applications in the biomedical field. Moreover, our innovative laser-based spectral imaging approach can achieve on-chip spectrometer-less chemical fingerprint retrieval eliminating clinically incompatible, complex, and bulky instrumentation requirements. The long-term goal of this project is to develop a rapid, label-free, portable, and non-invasive cancer detection platform based on sensitive and accurate chemometric liquid biopsy and machine learning-aided discrimination modalities. The overall objectives in this application are to (i) determine a potent metasurface design that can robustly extract chemical fingerprint information from a complex biosample matrix, (ii) identify optimized design parameters for spectral imaging-based on-chip fingerprint retrieval (iii) establish measurement protocols and data processing pipeline (iv) identify a machine learning model by which sensitive and accurate sample discrimination can be achieved. In the short term, we will pursue two specific aims: 1) develop novel engineered metasurfaces for sensitive and specific spectrochemical biofluid analysis and demonstrate spectrometer-less on-chip chemical fingerprinting 2) Test and validate the platform using biofluids from an ovarian cancer patient cohort and non-cancer controls. Our proposed approach is innovative because it catalyzes the state-of-the-art laser-based infrared spectral imaging technology with powerful nanophotonic tools to enable its impact in biomedical diagnostics and address an unmet medical need. In addition, the proposed interdisciplinary project is significant because it is expected to develop a non-invasive and accessible health screening platform that can ultimately impact the clinical management of cancer and the survival outcomes equitably among diverse populations.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY/ABSTRACT Currently approved stroke therapies include underutilized, time-limited, systemic thrombolysis and mechanical recanalization options. Therefore, there is a need for more affordable and flexible approaches to treatment that can be disseminated widely. One approach includes targeting robustly expressed innate pathways such as the Toll-like Receptor (TLR) signaling pathways activated after focal cerebral ischemia. There is a lack of knowledge regarding the timing and cellular specificity of downstream pathways activated by pathways such as TLR4 in astrocytes and other components of the neurovascular unit. In this proposal, we will determine the key downstream targets in astrocyte TLR4 signaling and the effect of astrocyte-specific TLR4 signaling on blood brain barrier permeability (BBB) and neurobehavioral outcomes following acute focal cerebral ischemia. We will use a clinically relevant, endogenous danger associated molecular pattern (DAMP), HMGB1, a known TLR4 ligand to determine the key downstream targets in astrocyte TLR4 signaling. We will also use in vitro models of ischemia, such as Oxygen Glucose Deprivation (OGD), to determine the TLR4-dependent downstream pathways activated by other DAMPs in astrocytes. Characterization of the downstream effectors of astrocyte TLR4 signaling has important implications for decreasing BBB permeability and secondary brain damage and improving outcomes after stroke. The overall hypothesis of this proposal is that stroke-induced, astrocyte-specific TLR4 signaling induces BBB disruption in the acute phase of stroke, and that inhibiting ischemia-relevant DAMP-TLR4 signaling in astrocytes will decrease BBB permeability following acute focal cerebral ischemia and improve behavioral outcomes. This central hypothesis will be tested in the following aims: Aim 1: We will determine the signaling pathways active in TLR4-reactive and TLR4 non-reactive penumbral astrocytes using a model of middle cerebral artery occlusion (MCAO) and transcriptomics following acute cerebral ischemia Aim 2) Using HMGB1 stimulation of cultured astrocytes and an in vitro model of cerebral ischemia, OGD, we will identify downstream targets of TLR4 signaling in astrocytes via Western blot and phosphoproteomics. Aim 3: Using mice with inducible, astrocyte-specific deletion of TLR4, we will determine the effect of astrocyte-specific TLR4 deletion on BBB permeability and neurobehavioral outcomes following MCAO. At the end of these studies, we will have a better understanding of the molecular mechanisms that underlie TLR4 signaling in astrocytes. Results from these studies will lay the foundation for the development of novel therapeutics that can decrease brain damage after stroke.

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), the etiological agent of coronavirus disease (COVID-19), has spurred an unprecedented global pandemic. Infection surges necessitate new therapeutic agents that are effective against a rapidly changing virus. Antivirals that inhibit viral entry into host cells have proven effective against other viruses with similar mechanisms of pathogenesis. The long-term objective of this highly collaborative proposed research is to develop potent peptides inhibitors of SARS-CoV-2 infection that operate by blocking structural rearrangements of the spike protein required for viral entry into host cells. For SARS-CoV-2 infection to occur, the viral surface spike (S) protein, a homotrimer, rearranges to form an energetically favored postfusion state. In this postfusion conformation, two helical domains, the N-terminal (HRN) and C-terminal (HRC) heptad repeats, associate to form a 6-helix bundle (6HB). Peptides derived from the HRC can inhibit formation of the 6HB, and thus SARS-CoV-2 infection. However, conventional peptides, composed entirely of α-amino acid residues, are highly susceptible to proteolytic degradation, which necessitates frequent and high dosing. The Gellman lab, in collaboration with virologists Prof. Anne Moscona and Prof. Matteo Porotto at Columbia University, has demonstrated that site- selective incorporation of backbone modifications, in combination with cholesterol conjugation, can decrease proteolytic sensitivity while maintaining high antiviral potency. Our team recently found that such lipopeptides can inhibit SARS-CoV-2 infection in biological assays and animal models, and that these inhibitors are effective against SARS-CoV-2 variants, SARS-CoV-1 and MERS. Building on this foundation, my proposed project seeks to develop lipopeptides containing backbone modifications that display high antiviral potency and resist proteolysis. Aim 1 will produce potent inhibitors of SARS-CoV-2 (and other coronaviruses) that contain backbone modifications and resist proteolysis. Aim 2 will evaluate the stability of 6HB formation between inhibitor candidates and the native SARS-CoV-2 HRN. Aim 3 will elucidate critical structural interactions between the HRC mimics and the native HRN. My hypothesis is that site-selective incorporation of backbone modifications into HRC-based designs will increase both antiviral activity and half-life in vivo, improving therapeutic efficacy. The proposed research, which will be conducted under the guidance of Prof. Sam Gellman at the University of Wisconsin, will provide me with experience in macromolecular X-ray crystallography, molecular design, protein engineering & expression, and virology. Through structure-guided engineering and sophisticated and multi-pronged assay implementation, these efforts could generate effective pan-variant therapeutics for COVID-19.

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY Quorum sensing (QS) is the process by which bacteria of the same species coordinate behavior at a high population density. In many pathogenic bacteria, QS systems are used to regulate virulence. This proposal focuses on the accessory gene regulator (agr)-type QS systems found in a several Gram-positive pathogens, including Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, and Clostridioides difficile. Agr-type QS systems contain four proteins, AgrA-D, that together produce and respond to an autoinducing peptide (AIP) signal. I will study the two proteins, AgrB and AgrD, that are responsible for signal biosynthesis. In this pathway, the peptide precursor AgrD is processed by AgrB and an extracellular protease to produce the AIP signal. In Aim 1, I will mutate the AIP region of S. aureus AgrD, testing to see if the AIP signal is still produced. Through iterative rounds of mutation, I will determine where in the peptide and to what extent variation is tolerated. Imbedded within this approach to understand the basic mechanisms of AIP processing is two additional goals, one of which has already been realized. First, I have tested the ability of the native system to produce non-native AIP analogs that act as potent inhibitors of S. aureus QS and have demonstrated that two highly potent pan-group inhibitors of S. aureus QS can be biosynthesized using my system. Second, by uncovering which residues of the AIP signal can be mutated without a loss of processing, I can test the non-native AIP analogs produced for their ability to inhibit QS in S. aureus, potentially discovering new and more potent inhibitors. For Aim 2, I will engineer a non-pathogenic bacterium to constitutively express the QS inhibitors biosynthesized in Aim 1. Then I will test the probiotic strain’s ability to prevent S. aureus pathogenicity in a Caenorhabditis elegans model. Aim 3 will continue to study the processing of AgrD but will switch the focus to investigating the final step, wherein an extracellular protease cleaves the AgrD peptide to yield final AIP signal. One mystery of this process is that AIP signals, even those within a single species, often differ in their proteolysis site. To discover what drives this variability, I will make targeted mutations to AgrD and compare proteolysis sites for the native AgrD sequence to mutant sequences. Applying this knowledge, I will then biosynthesize designer AIP analogs that combine features from two or more native AIP signals. Together these three aims will significantly increase our understanding of AIP biosynthesis and provide a novel pathway to valuable chemical tools for inhibiting agr-type QS systems in major human pathogens.

NIH Research Projects · FY 2025 · 2023-02

Project Summary The overarching theme of this proposal is to apply robust and reliable reaction mechanisms to improve the efficiency of forming C–C bonds adjacent to alcohol or amine functionality. Among the most common strategies to these products is the addition of a Grignard reagent (or other carbon nucleophile) to a ketone or ketimine electrophile, but the broad applicability of these methods is limited by (1) the poor compatibility of organometallic nucleophiles with many functional groups common to drug scaffolds and (2) significant limitations inherent to migratory insertion mechanisms, such as sluggish or undesired reactivity. The central hypothesis of this proposal is that α-heteroatom radical generating processes can be implemented in tandem with nickel cross-coupling for C–H functionalization to install hindered stereocenters. The emergence of nickel catalyzed cross-electrophile coupling strategies has provided novel and complementary reactivity to traditional cross-couplings and the controlled generation of organic free radicals is central to the development of new methods in this area. Intramolecular hydrogen atom transfer (HAT) mechanisms proceed at a higher rate than nickel capture of alkyl radicals, and should be an appropriate approach to generate the radical. The Specific Aims of this proposal are: (1) development of a 1,5-HAT and nickel C–C bond forming cascade to synthesize tertiary alcohols from readily available secondary alcohols and a traceless auxiliary group; (2) a strategy in which α-heteroatom radicals formed through 1,5-HAT are intercepted for nickel facilitated C–C bond formation to prepare functionalized amino alcohols and amino acids; and (3) an approach for Csp3–Csp3 coupling of α-heteroatom radicals to primary radicals is presented, in which cross-selectivity should be driven by radical stability differences. All three of these Aims can be adopted into enantioconvergent reactions by employing chiral ligands to generate stereocenters, which is highly desirable in the context of pharmaceutical and natural product synthesis. The development and synthesis of drug candidates is limited by the reactions available to make them, and the strategies described in this proposal will facilitate Csp2–Csp3 and Csp3–Csp3 bond formation at α-heteroatom carbon centers, an important motif found in pharmaceuticals and bioactive natural products. The continued improvement of catalytic conditions that employ a large pool of coupling partners to prepare sterically congested alcohols and amines will be of significant interest to both academia and industry, and will enable broader chemical space accessible in future drug discovery endeavors.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY/ABSTRACT Advances in imaging and fluid-based biomarkers of Alzheimer’s disease (AD) including amyloid (A), tau (T) and neurodegeneration (N), allow detection of underlying disease pathology decades before the onset of dementia. Despite over a decade of AD biomarker research, the field still lacks the ability to accurately predict if and when individuals with preclinical AD (those with biomarker detectable pathology in the absence of cognitive symptoms) will experience dementia. Identifying factors that slow or quicken this preclinical timeframe is needed to improve dementia risk prediction for preclinical AD patients and to inform optimal treatment windows for clinical trials aiming to slow or prevent cognitive decline and impairment. Until recently, studying these factors was precluded by observing different people for short periods of time that began studies in different disease stages with no way to identify when disease began for individual participants. Our team developed and validated new methods that provide individualized estimated amyloid onset age (EAOA) from amyloid biomarkers. EAOA can be used to rearrange biomarker and clinical observations along an AD-specific timeline (i.e., an Amyloid Clock) anchored to the start of preclinical AD. This project will apply this novel approach to existing data from that Washington University Knight ADRC, the Wisconsin Alzheimer’s Disease Research Center, the Wisconsin Registry for Alzheimer’s Prevention, the, the Mayo Clinic Study of Aging and the Alzheimer’s Disease Neuroimaging Initiative to investigate factors across cohorts that influence the timing and trajectories of AD biomarkers and dementia. This study was initiated based on our preliminary findings showing considerable differences between individuals and cohorts regarding 1) when amyloid onset occurs, 2) the time between amyloid onset and dementia onset, and 3) factors that affect AD biomarker and dementia trajectories in AD. In addition, studies from our center and others have begun to link AD pathology, change in brain volume, and changes in cognition to social determinants of health (SDoH) like neighborhood disadvantage. However, possible links between SDoH and the timing and trajectories of AD biomarkers and dementia are not well-understood. Our hypothesis is that observed individual and cohort differences in AD trajectories are due to a combination of demographic, environmental, sociocultural, and biologic factors, and study design and sample composition. We will test this overall hypothesis in three specific aims: 1) identify common factors across multiple cohorts that influence the timing and trajectories of ATN biomarkers; 2) identify common factors across multiple cohorts that affect the time from amyloid onset to dementia; and 3) explore inter-cohort differences in AD biomarker and dementia trajectories. This study will leverage existing data in several well-characterized studies to provide new insights into mechanisms that explain when preclinical AD begins and how long this preclinical phase lasts. This is expected to improve AD dementia risk prediction for individuals and identify optimal windows for disease modifying and prevention therapies.

NIH Research Projects · FY 2025 · 2023-02

PROJECT ABSTRACT Placental pathologies stem from poor early placental development characterized by shallow invasion of trophoblasts, and are also associated with aberrant expression of miRNAs belonging to the primate-specific chromosome 19 miRNA cluster (C19MC). C19MC miRNAs are thought to have roles in trophoblast invasion and migration, however, their role(s) in primate embryonic and early placental development is not well-defined. I hypothesize that C19MC miRNAs will have primate-specific roles in trophectoderm lineage specification and early development of the primate placenta. The overall objective of my proposal is to determine the role of these miRNAs in primate trophoblast lineage specification and placental development in a rhesus macaque model, and reveal gene networks regulated by miRNAs in primate placentation. Towards this objective, I propose three Specific Aims. Specific Aim 1 (K99-phase). To define the expression of miRNAs in the primate embryo and trophoblast stem cells (TSC). This aim will establish the miRNA signature during embryo development through specification of the trophectoderm lineage and determine whether these miRNAs are expressed in a stage- or cell-type-specific manner. Specific Aim 2 (K99-phase). To determine the functional role(s) of C19MC members in TSC and differentiated trophoblast function. This experiment will directly overexpress C19MC miRNA members in TSC to identify genes and pathways regulated by these miRNAs, assessing their functional roles. Specific Aim 3 (R00-phase). To use genome editing strategies to globally perturb the C19MC and evaluate the impact of aberrant C19MC miRNA expression on embryo development and primate placentation. This Aim will 1) repress and overexpress C19MC cluster expression utilizing genome editing tools in TSC, 2) apply C19MC genome editing to embryos to evaluate the role of cluster expression on primate preimplantation embryo development, and 3) determine the impact of embryonic genome editing on trophectoderm function and differentiation in an in vitro implantation culture model. I have recently developed macaque TSCs with methods described by Okae et al. (2018), and have experience with rhesus IVF to derive embryos for genome editing and in vitro implantation experiments. TSC and embryo resources will be used to define miRNA and mRNA expression during embryo development and trophoblast lineage specification and identify miRNA-regulated gene networks in early placentation. Overall, the proposed research will establish a basic understanding of miRNA expression and miRNA target gene regulation in the embryo to placenta transition. Ultimately, we envision that the nonhuman primate model will allow us to extend these approaches to transfer of edited embryos to recipient dams, and to develop in vivo strategies to directly target the placenta for modification of miRNA expression for experimental and therapeutic purposes.

NIH Research Projects · FY 2026 · 2023-02

ABSTRACT Coccidioidomycosis is a re-emerging infection that NIH has prioritized for vaccine prevention. An experimental vaccine of live attenuated spores is highly protective after intranasal delivery. We propose to study this vaccine to define mechanisms by which lung epithelium regulates durable mucosal T cell immunity. Resistance against inhaled microbes is thought to reside within tissue-resident memory (TRM) cells, but little is known about how this intranasal vaccine induces lung TRM. Our preliminary data reveal that the vaccine elicits protective, Coccidioides endoglucanase 2 (C-Eng2) specific CD4+ T cells and that bronchiolar club cells and Ca++ calcineurin signaling in the cells are needed to mobilize inflammatory and T cells in response to vaccine. We also find that Microfold (M) cells descend from the bronchiolar club cells and facilitate T cell priming in response to the vaccine. From these preliminary data, we hypothesize that bronchiolar club cells and M cells regulate mucosal cellular immunity in response to intranasal vaccine. To test this hypothesis, we have created innovative tools: (i) transgenic mice to deplete epithelial cell subsets or their products to further define their role in inducing immunity; (ii) C-Eng2 specific tetramers to track and analyze protective CD4+ T cells and TRM in C57BL6 mice; and (iii) methods to isolate and culture human lung epithelial cells to translate results from mice to humans. We propose three aims to test our hypothesis. In Aim 1, we will elucidate early stages of the inflammatory response to intranasal vaccine regulated by bronchiolar club cells and M cells; in Aim 2, we will identify lung epithelial cell receptors - dectin-1, DUOX1 and DUOXA1 - and downstream PLCG2 that may sense intranasal vaccine and signal via Ca++ and calcineurin to mobilize mucosal immunity; and in Aim 3, we will define mechanisms of vaccine-induced durable mucosal immunity by studying lung TRM and the regulatory role of lung epithelium. In sum, we address the unmet need of vaccination against coccidioidomycosis. Our work is significant as it will define mechanisms by which a promising vaccine establishes T cell immunity at the lung mucosa. Results will identify tactics useful for other vaccine immunogens given intranasally, including subunit vaccines. The work will define correlates of immunity needed to advance this attenuated vaccine or next generation subunit vaccines against this high priority pathogen. The work will be done with state-of-the-art, cutting-edge tools. Our team of PI and Co-I’s will let us translate results from mouse to human, with tools and reagents for human lung epithelium.

NIH Research Projects · FY 2026 · 2023-02

ABSTRACT Carbohydrates are ubiquitous and play a vital role in many important biological processes. The development of efficient and selective chemical methods for the synthesis of carbohydrates and glycoconjugates is necessary to understand the specific roles of carbohydrates and for therapeutic development. The most prevalent functionality in carbohydrates is the hydroxyl group. There are two fundamental challenges in carbohydrate synthesis associated with the hydroxyl group: 1) site-selective functionalization of one hydroxyl in the presence of many other seemingly identical hydroxyls, and 2) stereoselective glycosylation. One long-term goal of this program is to develop methods to address these challenges and improve the efficiency and selectivity for carbohydrate synthesis. In the next five years, we will develop methods that can site-selectively functionalize hydroxyl groups, such as acylation, alkylation, and sulfation, in various minimally protected or unprotected glycosides in a predictable and general manner. We will also develop methods that can site-selectively remove protecting groups in carbohydrates. The directing groups that are site-selectively installed in carbohydrates will also allow us to access various types of glycosidic linkages stereoselectively. These transformations can significantly improve the efficiency and selectivity for the synthesis of carbohydrates. The other long-term goal of this program is to prepare carbohydrates and glycocongates with novel biological functions. Certain glycans on glycoproteins can be recognized by lysosome targeting receptors (LTRs), which then transport the glycoproteins to the lysosome for degradation. To take advantage of this natural process, lysosome targeting chimeras were recently reported for the degradation of disease-associated extracellular proteins. These degraders are created by conjugating carbohydrate ligands of LTRs on the cell surface with ligands that can bind to the extracellular protein targets. The receptor-ligand interaction then triggers the internalization of the extracellular proteins through receptor-mediated endocytosis, which further induces the degradation of the endogenous extracellular protein targets in the lysosome. This new strategy complements existing targeted protein degradation methods, which largely focus on intracellular proteins. In the next five years, we will develop a series of carbohydrate-based ligands for LTRs that can be used for the degradation of various extracellular disease associated proteins. During our previous studies, we recognized the enormous potential of transition metal catalysts and chiral organocatalysts in carbohydrate synthesis and the unique utility of glycoconjugates in cell-type selective targeted protein degradation. In the next five years, we will continue developing novel methods for the synthesis of carbohydrates and glycoconjugates, studying their applications in targeted protein degradation, and pioneering new research directions.

NIH Research Projects · FY 2026 · 2023-01

Project summary/abstract COVID-19 revealed weaknesses in respiratory pathogen surveillance. Tools to detect SARS-CoV-2 were initially limited. Now that SARS-CoV-2 is prevalent, schools and other congregate settings struggle to perform systematic surveillance. As influenza virus and other respiratory pathogens return to co-exist alongside SARS-CoV-2, pandemic fatigue undermines risk mitigation strategies that rely on behavior modification, creating an unprecedented risk of illness-caused absenteeism in schools. We hypothesize that frequent air sampling in K-12 schools is a simple, inexpensive, and accurate sur- veillance strategy for identifying when schools and the communities where they are located are at high risk for SARS-CoV-2 and influenza virus transmission. In this study, we will place an air sampler inside each of 16 K-12 schools and collect continuous air sam- ples twice weekly throughout the school year. We will test air samples for SARS-CoV-2 and influenza vi- rus, the two leading causes of respiratory virus absenteeism. A key question is whether detecting these viruses in air samples is non-inferior to comprehensive individual testing of people with influenza-like illnesses. We are partnering with the Oregon (Wisconsin) School District which already performs in- school rapid diagnostic testing of symptomatic individuals. Within this unique setting, we can determine if the detection rates of SARS-CoV-2 and influenza are similar between air surveillance and diagnostic testing of individuals. If they are similar, this would suggest that simple, inexpensive, and anonymous air sampling programs could be the foundation of school-based sentinel virus surveillance programs. We will also determine whether detecting and sequencing SARS-CoV-2 in air samples forecasts broader community disease spread. Using SARS-CoV-2 diagnostic testing, wastewater, environmental, and mo- bility data alongside in-school air sampler detection and sequencing data, we will develop and iterate a model using air sampler data to forecast community transmission. We will extend this model to forecast the risk for influenza viruses that are not currently surveilled systematically. Successfully developing precision air sampler surveillance for SARS-CoV-2 and influenza virus in a large public school setting would provide a model for a nationwide surveillance program. Such a program would lead to safer classrooms, improve awareness of respiratory viruses that threaten specific commu- nities, and increase resilience to respiratory pathogens that threaten schools in the future.

NIH Research Projects · FY 2026 · 2023-01

Project Summary One of the barriers to therapeutic neural regeneration in humans is the absence of neural stem cells in most regions of the adult brain. Nonetheless, it would be advantageous to induce regeneration from resident cells. In addition, progress in the field would be accelerated if neural regeneration from resident cells could be investigated using a genetic model organism. Toward this end, we have developed a novel adult neurogenesis model in the Drosophila melanogaster central brain. We find that despite the absence of known neural progenitors, cells in the adult Drosophila central brain proliferate following injury, giving rise to both new neurons and new glial cells. Further, the new neurons project both axons and dendrites to specific target regions. We also observe functional recovery of behavioral deficits, suggesting that the new neurons integrate appropriately into neural circuits. Our results are paradigm-shifting because they suggest that resident brain cells can mediate neural regeneration. Here, we propose to utilize the model to investigate the signaling pathways and cellular mechanisms that regulate adult neurogenesis. Based on compelling preliminary data, our central hypothesis is that adult-born neurons are responsible for functional recovery from brain injury and that these neurons arise separately from adult-born glia. This hypothesis is supported by multiple lines of evidence from our ongoing work. We have identified, and are now investigating, genes uniquely upregulated during neural regeneration. The work proposed here will provide critical data about the molecular mechanisms that underlie that adult neurogenesis. Our work is innovative and has translational relevance because it shifts the focus of neural regeneration away from stem cell transplants and toward resident cell populations and it may lead to the identification of therapeutic targets for the stimulation of brain regeneration in humans.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY Post-translational regulation of histones and other chromatin-associated proteins is a major means by which gene expression is modulated during normal and malignant development. Polycomb group proteins are essential for proper development and are frequently altered in human cancers. The Polycomb Repressive Complex 2 (PRC2) functions in a collaborative chromatin-based crosstalk with PRC1 and H3K27me3 to initiate and maintain gene silencing. We have found that PRC2 catalysis of H3K27me3 is inhibited in pediatric gliomas by two suspected tumor drivers: histone H3 K27M and EZHIP. Inhibition of PRC2 activity by K27M and EZHIP can result in aberrant gene expression, cellular differentiation, and cell proliferation. Despite a substantial reduction in H3K27 methylation levels caused by EZHIP or K27M, our work has revealed residual H3K27me3 at CpG islands near promoters of known and suspected tumor suppressor genes. Furthermore, evidence suggests that this residual PRC2 activity plays a critical role in supporting tumorigenesis. This proposal seeks to leverage and extend our preliminary findings to define the mechanisms by which K27M and EZHIP misregulate PRC2 to promote tumorigenesis by employing a multi-disciplinary approach that integrates biochemical, genetic, and genomic methods. Specifically, we will (1) define the role of aberrant PRC2 activity in promoting K27M and EZHIP-containing tumors, (2) determine the mechanism of targeting PRC2 to CpG islands in gliomas, and (3) define the mechanism of PRC2 inhibition by EZHIP. Expected results will help us formulate novel theories and provide crucial mechanistic basis underlying the pathogenesis by oncohistones. The knowledge generated in the course of this study will motivate future therapeutic efforts for treating pediatric gliomas.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY The objective of my laboratory is to characterize how molecular communication between bacteria and their animal hosts leads to specific and reproducible colonization. To accomplish this goal, the laboratory studies the Vibrio fischeri-squid system, in which the animal’s “light organ” is colonized exclusively by one bacterial species. This system is advantageous because bacteria colonize through the natural route of infection, all animals are colonized within three hours of bacterial inoculation into the seawater, the bacteria can be subject to detailed genetic manipulation, the precise site of infection can be imaged directly in the live animal host, and chemical analysis of the animal host enables detailed molecular investigations. Focusing on how squid are reproducibly colonized by the specific symbiont, to the exclusion of the millions of competing bacteria in seawater, has revealed key roles for bacterial aggregation and biofilm formation in promoting specific host- microbe interactions. Questions that our group is asking include: (1) How does a symbiont regulate a beneficial biofilm? Biofilms provide microbes with a protected environment in which they can act collectively and resist innate immune insults and antimicrobial compounds. V. fischeri elaboration of a symbiotic biofilm is required for entry into the host, providing an opportunity to study this process in the context of a natural host colonization model. Our past work identified BinK as a key negative regulator of biofilm formation and the planktonic-to-biofilm transition in the host. In this study, we examine how BinK interprets signals from the host and how that information is transmitted to V. fischeri. We examine mechanisms of signal transduction and seek to identify and characterize a ligand that regulates BinK activity. (2) What novel bacterial factors play critical functions in colonization processes? We have had success in applying global genetic approaches to identify bacterial colonization factors in V. fischeri. With a focus on novel and understudied bacterial genes for which the V. fischeri-squid system has the potential to elucidate protein functions, we identified a protein that has a substantial impact on biofilm formation and squid colonization. The protein is annotated as a putative RNA-binding protein, and we will characterize the molecular mechanisms by which this protein acts and determine how it impacts symbiotic biofilm formation. (3) How do small molecules influence microbiome specificity and colonization? We have begun to identify compounds that are present in the host and that are co-regulated with symbiotic behaviors. We will integrate genetic approaches to elucidate signaling pathways in the context of host colonization. A major strength of the V. fischeri-squid system is the ability to interrogate bacterial behavior in the intact animal host, and completion of these projects will enable a deeper understanding of the mechanisms underlying animal colonization by beneficial microbes.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY/ABTRACT This proposal presents a five-year research career development plan focused on improving diagnosis of post-traumatic sepsis using microbial DNA sequencing in plasma. I am an Assistant Professor of Surgery at the University of Wisconsin-Madison, with previous clinical and research experience in trauma surgery, patient- centric predictors of sepsis, and lab-based genomic analyses of plasma in patients with sepsis. I have assembled an outstanding mentorship team with expertise in infectious disease, cell-free DNA, genomic data analysis, and bioinformatics. The proposed training will enhance my development across the disciplines of translational research, laboratory and computational methods in genomics, and scientific writing. This will enable me to transition to research independence as a surgeon-scientist dedicated to improving outcomes for trauma patients with sepsis by developing better diagnostic tests based on microbial genomics. Sepsis is a life-threatening condition, estimated to cause one-fifth of all deaths worldwide. Approximately 25% of trauma patients develop sepsis during their hospital course and more than 20% of such patients die during the same hospitalization. Sepsis, if not recognized and treated early, may evolve into septic shock and multiple organ failure. Thus, the treatment of sepsis emphasizes timely initiation of antibiotics and source control in surgery/trauma patients. However, the diagnosis of sepsis in patients with traumatic injury is complicated because systemic inflammatory response to injury mimics sepsis. In the current paradigm, confirmation of sepsis is based on clinical signs, laboratory and radiographic findings, and medical scores which are usually obtained after sepsis onset. There are approximately 200 biomarkers of sepsis described in literature but nearly all such biomarkers rely on measuring inflammatory response, which is confounded in patients with trauma. Recent studies, including my preliminary work, have shown that metagenomic sequencing of plasma DNA can rapidly identify pathogens in patients with clinical suspicion of sepsis but these assays have not been investigated in trauma patients. To address this gap, I propose to develop and evaluate a metagenomic sequencing-based assay for analysis of microbial DNA shed into the blood. My central hypothesis is that quantitative analysis of microbial DNA levels in trauma patients can predict sepsis, rapidly identify the causative organism of sepsis, and help decrease the duration of antimicrobial treatment. Using prospectively collected plasma samples from trauma patients, I propose the following aims: 1) To determine if an increase in circulating microbial DNA levels is predictive of sepsis in trauma patients, 2) To assess concordance between causative organism identified using standard-of-care microbiology and detection of pathogen-specific DNA in plasma, and 3) To evaluate circulating microbial DNA levels in plasma as a biomarker for monitoring antimicrobial response.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Sympathetic stimulation of ventricular myocytes activates PKA, which phosphorylates L-type Ca2+ channels and phospholamban, among other substrates, to increase Ca2+ entry and SR Ca2+ uptake. This β- adrenergic-induced increase of intracellular Ca2+ (Ca2+ overload) is a natural and efficient mechanism to increase cardiac performance, inasmuch as the magnitude and force of cardiac contractions greatly depend on the amount of Ca2+ supplied to the myofilaments. However, Ca2+ overload also increases arrhythmia vulnerability because it brings ryanodine receptors (RyR2) closer to threshold for spontaneous Ca2+ release (SCR). SCR during diastole activates the Na+/Ca2+ exchanger of the sarcolemma and generates a depolarizing inward current (delayed after-depolarization or DAD) that may trigger extemporaneous actions potentials. Ca2+ overload and its subsequent SCRs are indeed the primary events that evolve in malignant cardiac arrhythmias, as observed in Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT) and quite possibly in some forms of atrial fibrillation, long QT, and heart failure. To prevent these Ca2+-dependent arrhythmias, an emergent group of RyR2 blockers aim to stop SCR, despite the fact that the key process that spawns SCR, namely, Ca2+ overload, may not be affected or in some cases may be even exacerbated by the RyR2 blockers. We have found that imperacalcin, a high-affinity, membrane-permeable selective agonist of RyRs, paradoxically decreases arrhythmia burden and prevents Ca2+-dependent arrhythmias in animal models of CPVT. We hypothesize that this unexpected and remarkable anti-arrhythmic effect of imperacalcin is due to a partial and controlled depletion of SR Ca2+ load, which decreases the propensity of SCR events and thus dissipates the main arrhythmogenic substrate in CPVT. Our research program will systematically and rigorously test this hypothesis at the molecular, cellular, whole heart and intact animal levels using established (mouse) and novel (rabbit) models of CPVT. The first specific aim will determine the defining structural and functional characteristics that confer to imperacalcin and other members of the calcin family their capacity to permeate membranes and their anti-arrhythmic properties. In the second aim, we will use native and modified calcins on intact cardiomyocytes, Langendorff-perfused working hearts and intact animals for a rationalized design of a novel group of RyR2 ligands capable of preventing Ca2+-triggered arrhythmias. These studies use animal models of CPVT to develop a novel paradigm for the treatment of Ca2+ overload-generated cardiac arrhythmias; results may be applied to other cardiomyopathies where controlled unloading of SR Ca2+ may be desirable.

NIH Research Projects · FY 2026 · 2023-01

Abstract Text Project Summary/Abstract To successfully colonize and establish themselves in the gastrointestinal (GI) tract, enteric pathogens must sense and respond to several microbiota and host derived signals to properly regulate expression of their virulence repertoire. An important signal within the GI tract is the host neurotransmitter norepinephrine (NE) and/or epinephrine (Epi), which have important functions in intestinal physiology. There is also an important relationship between neurotransmitters and the microbiota, because it modulates the levels of active Epi and NE in the gut lumen. The host conjugates Epi/NE to glucuronate to inactivate it. The microbiota encodes glucuronidases (encoded by the uidA gene) that deconjugate glucuronate from Epi/NE, increasing the levels of free biologically active Epi/NE in the lumen. We identified the first two bacterial adrenergic receptors, QseC and QseE. The enteric pathogen enterohemorrhagic E. coli (EHEC), exploits adrenergic signaling through QseC and QseE to carefully regulate expression of its virulence genes to promote optimal colonization of the gut. Importantly, EHEC virulence gene regulation intersects with the ability to sense glucuronate, linking deglucuronidation of Epi/NE by the microbiota to Epi/NE sensing at another level of inter- kingdom signaling. We identified ExuR, the sensor for glucoronate, as an important regulator of EHEC virulence gene expression. The linking of Epi/NE sensing through QseC and QseE, with glucuronate sensing through ExuR, ensures the precise control of virulence gene expression by EHEC. Indeed, Citrobacter rodentium (extensively used as a surrogate EHEC model for murine infections) qseC, qseE and exuR mutants are attenuated for murine infection, highlighting the important role of this signaling system in EHEC pathogenesis. Another neurotransmitter system, the endocannabinoid system is also intertwined with the gut microbiota. The endocannabinoid 2-Arachidonoylglycerol (2-AG) is sensed through QseC, preventing QseC function, and decreasing expression of virulence genes in EHEC and C. rodentium. Moreover, Epi and 2-AG antagonize each other at the level of QseC sensing. The levels of 2AG decrease from the small intestine to the colon, oppositely from the levels of Epi/NE. Given that EHEC colonizes the colon, the interplay between the adrenergic and endocannabinoid systems, likely has a key function in the biogeography of this pathogen within the GI tract. Accordingly the specific aims of this proposal are: Specific Aim 1: Investigate the relationship among the adrenergic and glucuronate signals in EHEC pathogenesis. Specific Aim 2: Investigate the role of the endocannabinoid system in EHEC pathogenesis.

NIH Research Projects · FY 2026 · 2023-01

Despite widespread clinical use, the theoretical framework by which to understand safety of electrical stimulation through implanted electrodes is surprisingly limited. Most of our current understanding of stimulation safety was phenomenologically determined in the 80s and 90s using very limited electrode geometries, materials, stimulation systems, and stimulation locations. Current benchtop testing of electrode safety to support submissions to the Food and Drug Administration (FDA) is predominantly focused on identifying the applied charge density that drives the hydrolysis of water at the electrode/electrolyte interface. In this proposal, we seek to validate, optimize, and distribute a benchtop testing framework that more accurately predicts chronic in-vivo safety issues. This framework is extensible to coated microelectrode designs, including high-density and/or thin-film arrays, as well as to novel stimulation waveforms – both of which are critically enabling for next-generation minimally invasive neuromodulation therapies.

NIH Research Projects · FY 2024 · 2022-12

PROJECT SUMMARY/ABSTRACT Suicide rates have risen sharply over the past 20 years1. There is a need to more precisely identify proximal risk indicators for the development of near-term suicide risk in order to effectively intervene. Studies utilizing ecological momentary assessment (EMA) to collect data at several intervals per day have demonstrated that suicidal ideation (SI) and proximal risk factors change rapidly across the course of the day2. However, prior EMA studies examining SI dynamics implement stable assessments, with intervals of several hours between SI assessments across the duration of a study period3 for all participants. This one-size-fits-all approach to SI assessment fails to capture the nuanced within-person variability of the timescale of the development of acute suicide risk. In turn, we lack even a basic understanding of within-person variability in the time varying relationship between SI and its proximal risk factors. The proposed study aims to address the limitations of current assessment approaches in proximal suicide risk research through the development of a personalized, adaptive time sampling system. The specific objectives of the proposed research are to: (1) develop a novel, adaptive time assessment system that more efficiently and accurately identifies when an individual is at highest risk for SI; and (2) advance the understanding of SI and its theoretically-informed proximal risk factors at finer timescales. Data collected according to varied timing schedules in the first phase will be used to train an algorithm that generates predictions of suicide risk, predictions that will be adaptively use to determine assessment timing during the second phase of data collection. Aim 1 is to develop the adaptive time assessment system, followed by assessing the predictive accuracy of the adaptive sampling system (Aim 2) and identifying variations in person-specific effects of the relationship between SI and theoretically-informed risk factors (Aim 3). The research team (PIs: Ammerman, Jacobucci; Co-I: Cheng; Consultants: Burke) has access to world-class expertise, with extensive experience in EMA data collection in high-risk samples, machine learning for suicide prediction, longitudinal data analysis, collecting and modeling continuous data streams, and the development of adaptive assessment platforms. To meaningfully reduce suicide rates, a more nuanced understanding of suicidal thoughts and associated risk factors is required. Our adaptive assessment platform will more efficiently assess suicidal thoughts and risk factors, allowing for a closer approximation of the true associations. Indeed, there is a need to identify near-term risk factors prior to suicidal thought occurrences to successfully deliver an intervention and prevent suicidal outcomes. These findings will support the successful implementation of just-in-time adaptive interventions through increased precision of suicide risk detection and targeted intervention timing. Given the grave personal and societal cost of suicide, this work has important public health implications.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY/ABSTRACT Adverse Childhood Experiences (ACEs) have become a significant public health concern in the U.S. ACEs are potentially traumatic events that occur to children under the age of 18, including all types of abuse, neglect, and exposure to household challenges (e.g., having lived with a household member with substance use disorder). Varying sources suggest that 50-60% of U.S. adults report at least one ACE, and 15-25% report three or more. These statistics are concerning because experiencing ACEs has been shown to negatively affect various aspects of adult lives, including health outcomes, relationships, and financial and social status. Despite this established research, the impact of ACEs has not been studied in the context of caregiving for aging parents. Extensive prior studies have identified a comprehensive list of risk and protective factors related to caregiver stress; however, these factors tend to be proximal with little attention given to distal or early life course factors such as ACEs. Relying on the life course perspective, the proposed research aims to examine the effect that ACEs have on the experience and outcomes of caregiving for aging parents. Using nationally representative data from the Midlife in the United States (MIDUS) studies, secondary data analyses will be performed to address the first two aims: 1) Describe the prevalence and characteristics of filial caregivers who experienced ACEs, and 2) Examine the effect of ACEs in the association between daily caregiving and short- and long-term health outcomes. The second aim particularly focuses on exploring caregivers’ physiological functioning using daily cortisol levels to successfully quantify stress effects associated with filial caregiving. In addition, qualitative research will be conducted to address the third aim: 3) Explore caregivers’ experience of ACEs and illustrate whether and how ACEs affect their caregiving experience in terms of stress sources and the strategies they use to cope with caregiving stress. This K01 award would provide Dr. Jooyoung Kong with the training required to become an independent scholar and leading expert in later-life family relationships and caregiving for adults who experienced childhood trauma and adversity. The proposed training plan would allow Dr. Kong to receive instruction and mentorship toward meeting the following career goals: 1) Increase substantive knowledge in psychobiology; 2) Obtain advanced training in quantitative research methods and analysis; and 3) Gain advanced training in qualitative research methods and analysis. The K01 award will lead to an R01 grant application that will propose to conduct primary data collection to further investigate caregivers with histories of ACEs informed by Dr. Kong’s newly acquired substantive knowledge and methodological skills. Ultimately, this research will inform novel programs and policies to improve the health and well-being of family caregivers whose roles are becoming more significant in the current aging society.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY/ABSTRACT This K23 application proposed a career development plan to help Dr. Beth Fields establish an independent research program that focuses on optimizing hospital-based care and outcomes for patients living with Alzheimer’s disease and related dementias (ADRD) and their family member or friend care partners. She will train under the mentorship of a transdisciplinary group of senior scientists with research expertise in participatory human-centered design, conduct of clinical trials, and clinical ADRD care. She will continue working with her current mentors, Drs. Farrar-Edwards, Werner, and Shah, and Mr. Hetzel, all of whom have experience mentoring trainees. This will be complemented by content and mentoring expertise from Drs. Schulz and Gilmore-Bykovskyi. Collectively, this team will provide an outstanding training environment that will allow Dr. Fields to fill critical gaps in her knowledge and skill set relating to the study of hospital-based care processes and outcomes for patients living with ADRD and their care partners. Her training goals are to develop skills in (1) participatory human-centered design principles, (2) hospital-based ADRD care delivery, (3) clinical trial design and statistical analysis, (4) ethical and regulatory standards in ADRD research, and (5) professional skills in team science and scientific leadership. Achieving these goals will strengthen her scholarly activities, establish important collaborations, and acquire critical data that will ensure her successful transition to independence. To this end, Dr. Fields’ proposed research plan builds directly on her prior work developing and validating the Care Partner Hospital Assessment Tool (CHAT). Guided by the widely used and effective decision-support model of Screening, Brief Intervention and Referral to Treatment (SBIRT), the CHAT applies a sequential screening and referral pathway that 1) identifies care partners and their preferences for inclusion in the patients’ hospital care and 2) tailors referrals to address their stated preferences and unmet needs for post-discharge preparedness. The SBIRT model was designed to adapt flexibly to different health conditions and contexts, thus enabling the adaptation of the CHAT to facilitate the inclusion and preparation of care partners of patients living with ADRD. Therefore, the purpose of this proposal is to adapt CHAT for care partners of hospitalized patients living with ADRD (CHAT-AD) and evaluate its feasibility and potential efficacy in a pilot randomized clinical trial. Findings from this study, in combination with the career development plan, will enable Dr. Fields to launch an independent program of research that aims to (1) improve hospital-based care processes and outcomes for patients living with ADRD and their care partners, and (2) elucidate the essential caregiving role that so many care partners of patients living with ADRD assume.

NIH Research Projects · FY 2025 · 2022-12

Abstract There is strong clinical impetus to provide bilateral hearing early in life, not only for safety in navigating the environment, but also to maximize learning and socializing in mainstreamed settings. Although this topic is an active area of research, we lack critically important information on how to assess and interpret the impact of age at onset of deafness and auditory experience in children with bilateral cochlear Implants (CIs). Four groups of bilateral CI users and normal hearing (NH) participants in the same age ranges will be recruited to test hypotheses about the role of auditory experience and inter-implant delay in emergence of binaural hearing, functional listening and cognition. Here we propose novel studies aimed at gaining fresh insight into the role of auditory experience and inter-implant delay on outcomes. We will integrate perceptual and electroencephalographical (EEG) measures of binaural integrity (Aim 1), and functional listening with binaural cues (Aim 2). Further, cognitive measures of executive function (EF) are introduced to promote novel discovery of the association between binaural listening measures and EF (Aim 3). By synergistically combining these approaches, this project will be the first to provide urgently needed answers to timely and clinically critical questions regarding pediatric cochlear implantation. An important barrier to maximizing outcomes stems from engineering limitations whereby CIs are lacking binaural coordination. In addition, while speech envelope (ENV) cues are preserved in speech signals, cues that are significant for binaural hearing, namely temporal fine structure (TFS) cues, are not preserved in CI processing. We will systematically manipulate the control of ENV and TFS cues by using either research processors that allow exquisite control of timing of stimuli reaching each electrode or presenting stimuli to clinical processors and in free field, where stimulation is more akin to today’s CI processing. comprehensive investigation into outcomes in children with bilateral CIs, using perceptual, eye tracking and EEG measures at multiple levels of auditory processing. The stepwise assessment along the neural axis, from brainstem to auditory cortex, to cortical connectivity and whole brain coherence analyses, will enable us to understand which perceptual deficits are associated with abnormal neural processing. Cognitive measures are introduced to promote novel discovery of how abnormal neural development is related to EF, and if these effects show pervasive effects beyond auditory-based EF tasks. This information is important for understanding how the timing of bilateral CIs is related to a set of cognitive processes– regulating attention, memory, and controlling cognitive behaviors – necessary for success across academic, social, and daily living domains. We will harness the tests used in our studies to develop a clinical assessment toolbox that can be used by clinicians to assess binaural hearing abilities, and our findings will further identify which cognitive EF measures should be use along with auditory measures to clinically assess outcomes in children. Results will inform selection of design features in engineering CIs, with an emphasis on binaural processors, thus aiming to reduce the gap in performance relative to NH listeners.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT MIDUS has unprecedented opportunities to advance knowledge of risk and protective factors for cognitive decline as well as for Alzheimer’s Disease (AD) and Related Dementias (ADRD). Such potential stems from its comprehensive assessments from two national samples (Core, Refresher) of behavioral, social, psychological, and biological assessments from prior decades in the participants’ lives. Identifying markers of risk before disease symptomatology is foremost to AD/ADRD prevention science. Key aims are to: (1) Conduct additional waves of cognitive assessments on both national samples and obtain new measures focused on cognitive impairment. The Brief Test of Adult Cognition will be re-administered to ~ 4,000 adults (Core n = 2,062; Refresher n = 1,935), ages 25 to 95 at the last wave with key neuropsychological assessments of memory, speed, fluency, reasoning, and executive functioning. Global cognitive status will be assessed with the Telephone Interview for Cognitive Status and self-reported symptoms. (2) Conduct additional waves of emotion-related functional neuroimaging and psychophysiological assessments on Core and Refresher Neuroscience subsamples and obtain comprehensive measures of brain aging. Multimodal neuroimaging and psychophysiological data will be collected on ~ 450 adults (Core n = 215 longitudinal + 60 new; Refresher n = 115 longitudinal + 60 new). Longitudinal analyses will examine changes in affective chronometry of emotional processes, computed brain age, atrophy, white matter structural integrity and hyperintensities, microstructural complexity of dendrites and axons, and network connectivity and modularity. (3) Quantify AD and neurovascular disease burden and collect new ADRD-related plasma and neuropsychological measures and clinical assessments in the Biomarker subsamples. Conduct advanced molecular amyloid PET to identify individuals exhibiting amyloid accumulation indicative of AD neuropathic change, and neurovascular MRI to identify vascular diseases including vessel stiffness and oligemic tissue perfusion. Cross-validate plasma markers of amyloid beta (42/40) against amyloid imaging in participants who have both, and in conjunction with the MIDUS U19 collect aβ42/aβ40, p-tau181, ptau217, total tau, and neurofilament light (NFL) on the full biomarker samples (~ 1280 participants; Core n = 630; Refresher n = 647), thereby extending the reach of MIDUS to include ADRD biomarkers. In conjunction with the U19 application, these new measures will be used to test hypotheses regarding cognitive decline and the precursors of AD/ADRD and neurovascular disease via cumulative stress over 30 years and consider socioeconomic and race disparities and resilience. The protective influence of biopsychosocial resources, affective style, and lifestyle factors assessed over multiple prior waves of MIDUS will be examined in relation to early indicators to gain a better understanding of the relations of these factors to cognitive impairment and dementia. These new assessments and analyses offer ground-breaking science on mechanisms to advance prevention, including development of interventions and treatments for aging declines and dementia.