Oregon Health & Science University

NIH Research Projects · FY 2026 · 2025-07

PROJECT SUMMARY Tobacco use disorder is the deadliest substance use disorder accounting for nearly half a million premature deaths in the United States each year. Primary care clinics have the potential to reach the majority of the population with tobacco cessation assistance in the context of ongoing care, but uptake of evidence-based strategies is poor. Team-based and electronic health record-supported approaches to promote tobacco assessment and referral to cessation services (e.g. Quitlines) have been successful, but were significantly disrupted by the sequelae of COVID-19. Population outreach approaches using navigators and technologies such as patient portals have been effective in facilitating patient access to and engagement with care services and can improve health outcomes. Important gaps in understanding which outreach approaches effectively reach and engage low-income primary care patients with tobacco cessation support remain. We propose that using an outreach strategy focused on patients who were not assisted during a recent primary care visit will greatly improve the delivery of tobacco cessation support to patients. We examine two outreach strategies: a predominately person-based outreach navigator (ON) vs. a technology-based interactive patient portal (IPP). Patients that are not offered assistance during a primary care visit will be randomized into 3 groups: ON vs. IPP vs. usual care / no outreach group (NOC). Both implementation strategies will use an evidence-based approach to communicate the importance of cessation counseling and tobacco cessation medications for an effective quit strategy, offer patient choice among options, and facilitate referrals to tobacco cessation counseling and orders for medications using EHR tools. This study aims to: 1. Implement a tobacco treatment outreach intervention with ON vs. IPP vs. NOC to compare rates and correlates of receipt of cessation counseling and tobacco cessation medication orders. 2. Evaluate the effectiveness of ON vs. IPP vs. NOC on tobacco quit status. 3. Estimate and compare the costs and cost-effectiveness of ON vs. IPP. These aims will be accomplished with a three-arm randomized trial with 6000 patients (randomization ratio of 1:1:1) from 10 community-based clinics. EHR data and patient surveys will be used to assess the effect of the ON vs. IPP vs. NOC on patient reach and effectiveness outcomes and whether the approaches work equitably across important subgroups of patients. We will also assess the cost to implement and conduct each outreach approach and conduct cost-effectiveness analyses. For each Aim, qualitative methods will complement the hypothesis testing analyses to better inform observed differences in outcomes of the approaches. This study is well aligned with NIDA priorities and fills a critical knowledge gap of identifying effective and scalable strategies to improve tobacco cessation assistance among low-income primary care patients.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY / ABSTRACT This application requests funds to purchase a dual ultrasound-photoacoustic system needed for in vivo, non- invasive, longitudinal imaging of small animal models. The instrument, a Vevo F2 LAZR-X imaging system (FUJIFILM, VisualSonics), will support and enhance current NIH-funded projects, and promote innovative basic and translational projects at the Oregon Health & Science University (OHSU) and neighboring institutions. The Vevo F2 LAZR-X uniquely enables correlative ultrasound-photoacoustic imaging. Ultrasound allows to image organs with high resolution and assess organ function, as well as quantify tissue and blood flow velocities through Doppler ultrasound. Photoacoustic imaging (PAI) allows to determine tissue molecular content (including quantification of blood oxygenation) based on the light absorption properties of distinct molecules. Because photoacoustic and ultrasound signals are detected with the same transducer simultaneously, the system can generate images in which the molecular content (from PAI) is overlaid on high resolution (up to 30 μm) ultrasound images of the tissue or organ of interest. Moreover, since both ultrasound and PAI modalities are non-invasive and non-ionizing, the Vevo F2 LAZR-X system is ideal for in vivo longitudinal studies in small animal models. Technical advances incorporated into the Vevo F2 LAZR-X system, together with unique features, such as wide range of transducer frequencies (1 to 71 MHz) and laser wavelength range (680- 970 nm and 1200-2000 nm), as well as fast scanning (up to 10,000 frames per second when combined with EKV) make this instrument a must-have for the developmental, cardiovascular, musculoskeletal, and cancer applications of our Users. The proposed Vevo F2 LAZR-X system is at the vanguard and a top choice among competitive commercial ultrasound and photoacoustic imaging systems, and will uniquely enable basic and preclinical research at the forefront of discovery while advancing NIH funded research. The OHSU South Waterfront campus, where the Vevo F2 LAZR-X system will be located, is a hub for innovation in developmental, cardiovascular, musculoskeletal, and cancer research. Several of the Major Users in this application are developing photoacoustic contrast agents with the goal of translating into the clinic for improved cancer diagnosis and guided surgery intervention. Users are also seeking an effective imaging technique to longitudinally monitor cardiovascular performance during the lifespan. This proposed shared instrument will greatly benefit NIH-funded investigators at OHSU and beyond. The proposed system will replace two previous instrument models, which are currently out of manufacturer's maintenance and service (as of 12/2023) and out- of-order. There is no other dual ultrasound-photoacoustic system at OHSU or nearby. Thus, there is a dire need for the proposed system to boost developmental, cardiovascular, musculoskeletal, cancer, and photoacoustic contrast agent research at OHSU and Oregon. The Vevo F2 LAZR-X imaging system will be instrumental to advance our understanding of disease progression and treatment efficacy at different stages of life.

NIH Research Projects · FY 2026 · 2025-07

Project Abstract Many adult organisms, including humans, turnover a significant number of neurons during life, and many non- human organisms can regenerate substantial neural tissue following injury. New neurons in both cases are ultimately made from adult stem cells (ASCs), yet it is largely unknown how ASCs make the correct diversity of neural cell types at any given time while maintaining proper patterning and function in an otherwise healthy tissue. Understanding, at the single-cell level, how ASCs can access neural fates, make appropriate numbers and types of new cells, and then restore brain function is of fundamental importance in devising regenerative therapies for human neural injuries. However, in order to understand the mechanisms of successful neural regeneration, model organisms capable of both the biology and gene-function analyses must be used. To understand complex biology of adult neural regeneration we use complementary model organisms that can regenerate substantial parts of the nervous system following injury: the freshwater planarian and zebrafish. Planarians have the key advantages of being able to regenerate their entire brain following decapitation, as well as test gene function rapidly by RNAi during the regeneration process in vivo. Zebrafish are the best genetic vertebrate model of neural regeneration and have the advantages of live-imaging and the ability to recover from complete spinal cord transections. Only now do we have the tools in both systems to determine the conserved mechanisms of neural regeneration at the single-stem cell level. Despite much work from our lab and others, we understand little about stem cell heterogeneity or conserved stem cell programs of regenerative neurogenesis in either system. This proposal addresses these unknowns. In Aim 1, we will test four conserved transcription factors that we hypothesize drive the neural stem cell state in planarian brain regeneration. In Aim 2, we will test 57 novel factors that we have discovered in planarians that turn on specifically in neural-fated stem cells during brain regeneration, including two homologs of the conserved musashi family of RNA-binding genes. In Aim 3, we will determine the first roles of the musashi-1b gene in stem cells during spinal cord regeneration in zebrafish using conditional genetic strategies and live-clonal analyses. Similar to planarians, musashi-1b also turns on in zebrafish neural stem cells in response to injury and we will determine the RNA targets of musashi-1b in spinal cord regeneration. In total, this proposal will deliver conserved mechanisms of adult neural regeneration from stem cells that will be of broad relevance to designing regenerative therapies for human neural injuries.

NIH Research Projects · FY 2025 · 2025-07

Project Summary The International CD1-MR1 Workshop is the premier international conference focusing on the latest science regarding non-classical T cells, which are broadly defined by their lack of MHC restriction. This 4-day workshop has been held since 2000, and brings together T cell experts from around the world. Beyond MR1-restricted mucosa associated invariant T (MAIT) and CD1d-restricted NK T cells, this international forum describes new CD1- and MR1-restricted T cell populations participating in human inflammatory, autoimmune and infectious disease, as well as translational sessions describing new reagents to treat human T cell mediated diseases. The upcoming 2025 CD1-MR1 workshop will be held from September 9-12, at Oregon Health Science University in Portland, Oregon, and aims to further build the CD1-MR1 field by stimulating collaborations between early career and senior scientists in a setting conducive for networking. We have designed programs supporting visibility for trainees, as well as special lectureships for early and mid-career scientists. The recent establishment of an international CD1-MR1 Society will increase participation, further aided through travel awards to support students and postdoctoral fellows, social and scientific functions that are specifically designed to promote international interactions. Requested funds will primarily be allocated towards merit-based travel awards as well as reduced registration fees for early career investigators

NIH Research Projects · FY 2025 · 2025-07

Project Summary The external globus pallidus (GPe) is a central hub nucleus that processes information necessary for behavioral control by the basal ganglia, a group of interconnected forebrain nuclei. A wide range of behavioral disorders, including the formation of obsessive and compulsive habits, and the generation of psychotic, major depressive, and anxiety disorders are all thought to arise by dysfunctions of the basal ganglia because these nuclei process a wide range of information. Sensorimotor, associative, and cognitive information from many regions of the cortex and thalamus are integrated by projection neurons of the striatum, the main basal ganglia input nucleus. In order for the basal ganglia to control behavior, information from the striatum must then be integrated by the GPe. In the rodent, where the connectivity of the basal ganglia is best quantified, millions of striatal projection neurons (SPNs) form hundreds of millions of inhibitory synapses on only tens of thousands of GPe neurons. Each GPe neuron has two unique properties that may allow them to process an enormous amount of information: long dendrites and spontaneous somatic firing. The long dendrites of GPe neurons allow them to receive an enormous number of inhibitory striatal synaptic inputs, but also force these inputs to propagate along the dendrites before they can be integrated by the somata as changes in firing. The spontaneous firing of GPe neurons is driven by intracellular second messengers that activate two protein kinases that can enhance dendritic propagation. The goal of this proposal is to investigate how dendritic protein kinase activity (1) relates to the spontaneous firing activity of GPe neurons and (2) controls their integration of inhibitory striatal synaptic inputs. To address these questions, this proposal is divided into two aims. (Aim 1) Determine the relationship between dendritic protein kinase activity and spontaneous firing activity in GPe neurons. I will receive training in two-photon fluorescence lifetime microscopy to measure the spatiotemporal dynamics of dendritic protein kinase activity, while simultaneously measuring the spontaneous firing activity of GPe neurons using the cell-attached technique. (Aim 2) Determine how protein kinase activity controls the spike time responses of GPe neurons to dendritic input from the striatum. I will receive training in laser scanning photostimulation to selectively evoke dendritic striatal inputs, and will employ perforated patch clamp recordings to measure the effect of blocking protein kinase activity on the integration of these dendritic inputs. The proposed work will advance our understanding of information processing within the basal ganglia and will broaden my research approach. I will perform the proposed work under the mentorship of Dr. Tianyi Mao, a pioneer in the design and use of optical techniques to measure the spatiotemporal dynamics of protein kinase activity and the integration of dendritic synaptic inputs, at the Vollum Institute, a world-renowned research institute that has advanced our understanding of synaptic transmission and intracellular effector functions for the past four decades.

NIH Research Projects · FY 2025 · 2025-07

Project Summary Noise-induced hearing loss (NIHL) affects nearly one in four U.S. adults, making it the leading cause of acquired hearing loss, contributing to significant communication challenges and reduced quality of life. These challenges are particularly severe in complex listening conditions, such as when multiple sounds are present or in noisy backgrounds. These challenges often persist even with hearing aids. Currently, hearing-aid fitting is based on the audiogram, which measures hearing thresholds in quiet and does not fully address suprathreshold processing critical for real-world listening. Several neural mechanisms support complex-sound processing along the auditory pathway; how NIHL degrades these mechanisms is relatively well characterized in the auditory periphery, but not in the auditory cortex. Since the auditory cortex directly supports perception of complex sounds, it is critical to characterize how hearing loss affects cortical representations of complex sounds. A major impediment to understanding cortical processing in normal and impaired hearing has been the immense response diversity of auditory cortical neurons. We will address this limitation by using recent advances in large- scale electrophysiology and machine learning. In parallel, we will behaviorally assess categorization of sounds presented simultaneously, in noisy conditions – a task designed to capture real-world deficits due to NIHL. [K99 – normal hearing] Aim 1: We will perform large-scale single-unit recordings while presenting a massive natural sound corpus and use these data to identify the cortical manifold, which represents a low-dimensional representation of high-dimensional cortical activity. We will test whether manifold computations generalize across animals. Aim 2: Next, to reveal the neural basis of sound categorization, we will develop categorization models based on cortical single-unit data and validate these models using behavior. We will test whether neural models better match behavior than acoustic models not based on biology. [R00– NIHL] Aim 3: Using tools (cortical manifold and behavior) developed during K99, we will test whether frequency tuning broadens, inhibition reduces, and manifold degradation in noise correlates with behavioral deficits following NIHL. Aim 4: To directly test whether manifold-degradations underlie these perceptual deficits, we will design algorithms to restore the impaired cortical manifold to normal and compare their efficacy with traditional hearing-aid algorithms in improving neural representations and behavioral performance. These results will characterize, possibly for the first time, mechanisms by which NIHL impairs categorization of complex natural sound categories. Over the course of the project, the PI will be trained in cortical neurophysiology and modern machine-learning techniques, building on his existing expertise in peripheral neurophysiology, computation (modeling, signal processing), and translational neuroscience (noninvasive assays, hearing loss). This scientific and professional training will facilitate the PI's transition into an independent investigator position.

NIH Research Projects · FY 2026 · 2025-07

Improving access to opioid use disorder (OUD) treatment is critical to curb opioid overdose rates in the United States. Alarmingly, only 25% of adults with an OUD receive opioid agonist or antagonist treatment. In March of 2020, telehealth policies were enacted to increase access to OUD treatment, including federal regulatory agencies authorizing remote buprenorphine induction and prescription refills if a patient could be adequately evaluated and monitored via telehealth. Many insurers began reimbursing at the same rate for in-person, video and telephone visits. Prior to telehealth implementation, access and utilization of primary care-based OUD treatment were limited. Expansion of telehealth promoted access to OUD care that might not have been accessible otherwise and has the potential to reach patients with low rates of in-person primary care-based OUD treatment. Conversely, limited access to technology and low digital literacy could hinder the use of telehealth services in some populations (e.g., older adults, those living in rural areas). While telehealth has been shown to be as effective as in-person care in engaging and retaining patients on medications for opioid use disorder (MOUD), most studies were conducted within a few years of policy implementation. Significant gaps remain in our knowledge of how these changes have affected rates of and characteristics associated with OUD care as telehealth has evolved within health care settings and become a standard option for care delivery. Moreover, little is known about the patterns of telephone, video and in-person visits for OUD care over time or the multi-level characteristics associated with use of each modality. To date, no studies have comprehensively evaluated the impact of telehealth expansion on OUD care within community health centers (CHCs) which are at the forefront of the opioid epidemic. Our study uses almost a decade of CHC electronic health record (EHR) data (N>3.6 million patients in 35 states) from OCHIN, one of the nation’s largest safety-net networks with a single EHR. The data set includes over 90,000 patients with an OUD diagnosis, of whom more than 40% had an order for an MOUD. We link the EHR data set with community-level data and Datavant mortality data to: 1) Examine rates of OUD-related care (e.g., OUD diagnosis, MOUD orders, MOUD retention, behavioral health) in CHCs before, during, and after telehealth expansion; 2) Examine rates of OUD-related care in CHCs by visit modality (in-person, video, telephone) over time; and 3) Identify patient- and community-level factors that moderate the associations between visit modality and OUD care over time. This study uses a robust linked data set to conduct a comprehensive and rigorous examination of changes in the rates and moderators of OUD care among CHC patients seen before, during, and after the expansion of telehealth. Our evaluation informs future telehealth policy decisions and clinical guidelines, and determines what, if any, interventions are warranted within CHC settings to ensure access to OUD treatment for all individuals in need.

NSF Awards · FY 2025 · 2025-07

With support from the Environmental Chemical Sciences program in the Division of Chemistry, Professor Paul Tratnyek at Oregon Health & Science University and his students will study how the molecular size and shape of natural organic matter (NOM) influences its redox reactions with mineral surfaces. NOM plays significant roles in many biogeochemical/environmental processes and in almost all cases, these processes involve redox (oxidation-reduction) reactions that occur on the surfaces of particulate materials such as minerals and microbes. To characterize these reactions, the project will use a suite of electrochemical methods to measure electron transfer to and from samples of NOM with varying size and shape. The results will have a wide range of potential applications, including modeling of air pollution, management of agricultural soils, and optimization of drinking water disinfection. As well as graduate students, the project team will include interns from several nearby colleges that have strong programs for including undergraduate students in research. Reactions of NOM with the surfaces of minerals (and microbes) require sufficient proximity between the redox-active moieties in NOM and redox-active sites on the surface. This proximity is strongly dependent on the size and shape of the NOM, which can vary from discrete monomers to unfolded polymers to aggregates and other supramolecular structures like micelles. These effects can be studied electrochemically, since electrode response to NOM also is an interfacial process that is strongly dependent on NOM size/shape. The overarching technical objective of the proposed project will be to determine qualitative and quantitative relationships between the size and structure of NOM and its redox potentials, kinetics, and capacities using a complementary suite of electrochemical methods that allows for resolution of the many complex interactions involved. In the process, we expect to address several fundamental, cross-cutting issues regarding NOM redox chemistry, including that its bulk redox properties are the net effect of multiple redox active moieties that are linked by intra-molecular transfer, and that its capacity for electron exchange/storage reflects both labile and kinetically-limited components to varying degrees depending on operational factors. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2025-07

PROJECT ABSTRACT Reducing the global burden of low birthweight (LBW) remains a high priority for the World Health Organization (WHO). In Africa, malaria in pregnancy contributes to approximately 20% of LBW cases and affects nearly 12 million pregnancies every year. To curb the risk of malaria and LBW in Africa, the WHO recommends intermittent preventive treatment with sulfadoxine-pyrimethamine (IPTp with SP), a malaria chemoprevention strategy for pregnant women living in malaria-endemic settings. However, over the past two decades, widespread parasite resistance to SP has called for an urgent need to identify alternative antimalarials that could replace SP. While several antimalarials have been studied to date, the most promising candidate appears to be dihydroartemisinin-piperaquine (DP). Randomized controlled trials from our group and others have shown DP to be safe in pregnant women and far superior to SP in preventing malaria. Yet, these studies yield conflicting results on whether DP is superior to SP in preventing LBW. Mediation analyses conducted by our group confirm that the reason for this paradoxical finding is that SP, an antimalarial with known antibiotic and anti-inflammatory properties, improves LBW through mechanisms independent of its antimalarial activity (e.g., potentially through preventing sexually transmitted and reproductive tract infections, changing the gut or vaginal microbiome, and reducing maternal inflammation). Moreover, upon further investigation, the benefits of IPTp with either DP or SP appear to be context-specific, largely driven by the heterogeneity of the ‘non-malarial’ effects of SP between sites. Thus, in order to inform WHO on the optimal IPTp regimen, which may require a tailored approach for each setting, further evidence is needed to define the mechanisms driving the non-malarial effects of SP and for whom and where prevention of the malarial and ‘non-malarial’ mechanisms are most relevant. The objectives of this K99/R00 are to: characterize the mechanisms that mediate the effect of SP and DP on birthweight (Aim 1), assess the extent to which these mechanisms and other factors are causing heterogeneity between sites (Aim 2), and develop a model to estimate which antimalarial combination (either DP, SP, or a combination of DP+SP) would be the most optimal regimen for each unique epidemiological setting (Aim 3). Our research will leverage existing data from eight clinical trials conducted across ten study sites. The proposal will build on the applicant’s background in malaria, clinical trials, and epidemiology and include new training in: (1) the potential ‘non-malarial’ targets of SP affecting maternal and child health, (2) advanced computational statistics, (3) causal inference methods to target and tailor interventions. The training plan will be guided by an exemplary mentorship team who are experts in the field of causal inference, statistics, malaria, and maternal and child health. The combined research and training plan will competitively position the applicant for a successfully independent research career as an infectious disease epidemiologist focused on improving global maternal and child health policies.

NIH Research Projects · FY 2025 · 2025-07

Summary It is well understood how lipids are synthesized and metabolized in cells and that many lipids exhibit signalling functions to regulate cellular processes in a spatially and temporally defined way. The latter requires the build- up and turnover of lipid species in membranes either in a site-specific fashion or, alternatively, a directed form of lipid transport. This work aims to investigate the intracellular transfer of lipids from one membrane to another by several proteins that we discovered to be involved in lipid transport. In the previous funding period, we synthesized multifunctional lipid derivatives of five phosphoinositides and four common glycerophospholipids. These feature a photo-activatable protecting group (”cage”) to release the lipid derivative by light and a photo-crosslinking diazirine to covalently attach the lipid derivative to binding proteins. An alkyne group for click chemistry is useful for isolating lipid-protein conjugates or for determining the lipid location in cells by fluorescent tagging and microscopy. In published work, we identified specific lipid binding proteins for phosphatidylinositol 3,4,5-trisphosphate (PIP3), phosphatidylinositol 3,4-bisphosphate [PI(3,4)P2], and phosphatidylinositol (PI) via proteomic analysis. We then used siRNAs to block lipid transport and validated two hits that were required for transporting PIP3 and PI(3,4)P2: cytosolic MPP6 and transmembrane ATP11A. This R35 application proposes the continuation of work described in the application of R01 GM127631, namely the characterization of the lipid transport by the two above mentioned proteins (Project 1). This includes the purification and characterization of recombinant proteins and their functional mutants. We will use purified proteins to determine the 3D structure of MPP6 and its mutants by cryo-electron microscopy with and without crosslinked lipid derivatives. In Project 2, we will synthesize lipid derivatives featuring the photo-crosslinking diazirine closer to the membrane interphase to reach more transiently binding proteins such as those with a PH domain. Comparative proteomic analysis of the lipid interactomes will then be used to identify proteins involved in signalling with and without receptor stimulation. In Project 3, we will use multifunctional lipid derivatives to investigate the lipid interactomes of healthy and virus-infected cells. We recently discovered that an RNA virus infection leads to massive changes in the host cell lipidome. One exciting aspect is that one group of cellular phosphoinositides featuring a particular fatty acid composition is strongly up regulated. We will measure the lipid interactomes of flavivirus- and COVID-infected cells and identify targets crucial for viral infection and replication. Hits will be validated by protein knock-down and the effect on virus infection will be studied. Our unique lipid tools will help to better understand the lipid and lipid binding components of a viral infection.

NIH Research Projects · FY 2026 · 2025-07

ABSTRACT Head and neck squamous cell carcinoma is the sixth most common cancer worldwide with approximately 600,000 new cases diagnosed each year. Oral squamous cell carcinoma (OSCC) in particular, poses a significant health challenge due to its high prevalence and aggressive nature due to its propensity to invade bone. Therefore, elucidating mechanisms of bone OSCC growth and invasion that results in bone destruction is an urgent need in craniofacial and dental research. The calcified bone matrix and cells in the bone microenvironment make up a complex puzzle of variables that have been recognized to play important roles in the progression of oral squamous cell carcinoma towards bone invasion. This is especially due to the lack of in-vitro models that allow for a validated representation of these complex interactions as they happen in humans, in a manipulatable and systematic manner, which have remained virtually non-existent thus far. We have generated a tumor-bone interface on-a-chip model that includes all the players within the bone microenvironment, including mineralized matrix, osteoblasts, osteoclasts and the cells composing the bone vasculature. This system is highly manipulable, allowing for addition or deletion of the cell and matrix variables at play, which offer a unique opportunity to elucidate cause- effect relationships in oral cancer invasion into bone, and to validate next generation models of oral cancer. Here we propose the overarching hypothesis that (1) the reciprocal interactions between OSCC with bone, especially those mediated by osteocytes, regulate the process of oral cancer invasion and destruction of bone tissue, and (2) that our organ-on-a-chip model can replicate these interactions in a controllable, manipulable and validated manner. To address this hypothesis, we propose to (Aim 1) systematically manipulate, characterize and validate tissue microenvironment variables that have been linked to OSCC-bone invasion, (Aim 2) to elucidate and validate the biological mechanisms associated with these events in comparison to clinical samples, and (Aim 3) to test our system as a high-fidelity model to screen therapeutic effects targeting invasive OSCC. To ensure our model's clinical relevance and applicability, we will conduct rigorous validation through in- depth comparisons with clinical samples, assessing molecular signatures, histopathology, and therapeutic responses side- by-side. Our system represents a unique tool for academic researchers, pharma and regulatory bodies to assess drug efficacy and safety in a highly manipulable, high-fidelity human model for OSCC in bone.

NIH Research Projects · FY 2026 · 2025-07

Project Summary Hepatocyte transplantation has the potential to replace orthotopic liver transplantation for many liver diseases, especially genetic metabolic disorders. Cell therapy has significant advantages over liver directed gene therapy, including durability, the ability to administer repeat doses, and its applicability to multiple disorders, representing “one drug for multiple diseases”. However, significant obstacles to more wide-spread use of liver cell therapy remain. First, hepatocyte engraftment is rate limiting and it is difficult to achieve the replacement index needed for clinical benefit. Second, hepatocytes from organ donors are an allogeneic cell source and require immune suppression to persist. Here, we propose to overcome these barriers by using genetic enhancement of donor hepatocytes, making them immune stealthy and enabling selective growth expansion after engraftment. Three specific aims are designed to systematically explore multiple approaches to donor cell enhancement. In Aim 1, we will render donor hepatocytes resistant to acetaminophen (APAP) by knockout of Cypor. Two models of genetic liver disease (alkaptonuria and gyrate atrophy) will be used to test the efficacy of donor hepatocyte selection with APAP. In Aim 2, immune stealthy hepatocytes will be generated by knockout of class I HLA genes followed by class II HLA genes under inflammatory conditions. Gene-edited hepatocytes will be tested in allotransplantation models. In Aim 3, we will combine the manipulations to render donor hepatocytes immune stealthy and expandable. Fully enhanced allogeneic donor hepatocytes will be tested in a phenylketonuria disease mouse model. Together these experiments will lead to proof-of-principal that immune stealthy, expandable hepatocytes can overcome the major obstacles of therapeutic hepatocyte transplantation.

NIH Research Projects · FY 2025 · 2025-06

Project Summary There is a lack of longitudinal research examining neurodevelopmental trajectories spanning the fetal to postnatal period. The ability to characterize normative within-individual brain growth across this dynamic epoch is imperative to establishing the degree to which gestational measures can be used to predict neurodevelopmental abnormalities in individuals prior to later symptom onset. The goal of this proposal is to characterize the relationship between gestational and infant measures of neurodevelopment and determine the degree to which these measures are associated both structurally and functionally across the beginning of an individual's lifespan. The proposal leverages a dataset comprised of 229 serially imaged pre- and postnatal brain scans. Individuals received longitudinal structural T2-weighted scans beginning at mid gestation (postconceptional day 85) continuing through birth and into late infancy/early adolescence (postnatal 11 months) with the inclusion of postnatal diffusion-weighted imaging. Additionally, postnatal behavioral testing was conducted at one month of age. Aim 1 will characterize multiple cortical developmental indices (cortical folding, surface area, and volumetric expansion) across early development. Here, I test the hypothesis that in utero measures are associated with postnatal measures within-individuals using mixed effect modeling. Aim 2 will expand upon the previous aim to identify the degree to which gestational cortical macroscopic measures are associated with postnatal subcortical white matter maturation (fractional anisotropy and normalized T2-intensity), with the prediction that measures of gestational curvature will be associated with initial postnatal white matter organizational development. Finally, Aim 3 seeks to determine whether the developmental indices and relationships generated from the previous two aims explain individual variability in behavioral sensorimotor development. We hypothesize that gestational measures can alone predict composite measures of behavioral sensorimotor task performance and will be assessed using a Generalized Mixed Modelling approach. Together, these aims will provide a wide-ranging characterization of neurodevelopment, linking pre- and postnatal development measures longitudinally, which represent an under-studied and exceedingly important area in developmental research. The results generated from the proposed experiments will have implications for identifying the gestational measures best suited for early identification of neurodevelopmental abnormalities.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY The myelin sheath is a multilayered membrane generated by specialized glial cells called oligodendrocytes (OLs) that iteratively spiral their plasma membranes around axon segments in the central nervous system (CNS) of vertebrates. Myelination serves to increase the conduction velocity of action potentials, and OLs provide trophic support that is vital for neuronal survival. The importance of myelin in human health is underscored in diseases in which it is disrupted, like leukodystrophies and multiple sclerosis. Despite the importance of myelin, we lack a complete understanding of the molecular and genetic mechanisms that govern OL development and myelination; additionally therapeutic strategies to promote remyelination are sorely lacking. This proposal focuses on two critical regulators of OL development and myelination: Fbxw7 and Myrf. Loss of Fbxw7, a ubiquitin ligase component, in animal models leads to increased amounts of myelin formation, whereas loss of Myrf results in a lack of myelination. Fbxw7 targets Myrf for degradation, and we hypothesize that Fbxw7 functions to limit myelination in the CNS by acting, at least in part, by controlling Myrf levels. In this proposal, we will employ a synergistic combination of zebrafish and mouse models to: 1) define the cellular mechanisms by which Fbxw7 regulates OL development, myelination, and myelin homeostasis; 2) determine the role of Fbxw7 in remyelination; 3) Define the cadre of myelin-promoting Fbxw7 targets in OLs, with a particular focus on Fbxw7’s regulation of Myrf. These experiments will enhance our understanding of OL biology and may lay the foundation for future therapeutics that stimulate myelin repair in humans.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY With the most people ever in history currently living with HIV, stopping the HIV epidemic remains imperative. Combination antiretroviral therapy (ART) limits viral replication, but is not curative. Thus, there is an urgent need to design a functional cure via elimination of the viral reservoir. Five individuals, Timothy Brown (Berlin Patient), Adam Castillejo (London patient), Paul Edmonds (City of Hope patient), Marc Franke (Düsseldorf patient), and the New York City patient who has not released their identity were cured of HIV following leukemia-related, MHC- matched, allogeneic hematopoietic stem cell transplantation (alloSCT). How alloSCT mediates cure of HIV is unknown. Using our alloSCT model in Mauritian cynomolgus macaque (MCM), we have demonstrated that the allogeneic immune response can potently purge the latent reservoir. Therefore, we hypothesize that allogeneic T cell responses targeting minor antigens mediate HIV cure similar to how they mediate cure of leukemia. We propose here to define the allogeneic T cell responses in both the individuals who were cured of HIV, as well as the two MCM cured of SIV following alloSCT. In specific aim 1, we will define the allogeneic T cell responses and their targets in three of the individuals cured of HIV (Adam, Paul, and Marc), as well in a number of other cases where individuals underwent alloSCT and experienced a disappearance of HIV from the blood. In specific aim 2, we will define the allogeneic T cell responses and their targets in the two MCM cured of SIV post alloSCT, as well as in a further longitudinal cohort of SIV+ MCM that underwent alloSCT and experienced a decrease in the viral reservoir. In aim 3, we will adoptively transfer T cells expressing human or MCM allogeneic T cell receptors into HIV+ humanized mice or SIV+ MCM, respectively, to measure the safety and efficacy of allogeneic T cells for HIV cure. This work would expand our knowledge of the mechanism of HIV cure in the setting of allogeneic HSCT and establish a new therapeutic approach for HIV cure.

NIH Research Projects · FY 2026 · 2025-06

Summary Cannabis use disorder (CUD) affects at least 10% of the 193 million individuals that use cannabis worldwide. Cannabis is frequently promoted as a sleep aid despite conclusive evidence to support such benefits. Further, abrupt cessation of regular cannabis use is associated with sleep disturbances, often leading to reinitiating cannabis use. Thus, while the mechanisms by which cannabis regulates sleep remains unclear, the escalation and/or maintenance of hazardous levels of cannabis use likely involve a bidirectional (and mutually reinforcing) relationship between sleep and cannabis use. In this prospective mechanistic randomized control study, we will determine the impact of sleep restriction or extension on the amount of cannabis use and in turn the impact of this cannabis use on sleep continuity and sleep homeostasis, a pivotal component of sleep regulation. To examine this bidirectional relationship between sleep and cannabis use, as well as the hypothalamic-pituitary- adrenal (HPA) axis as a potential mediator, 60 participants (20 healthy controls, 20 with moderate cannabis use, and 20 with severe CUD) aged 21-29 years (a demographic with the highest prevalence of risky cannabis use and CUD) will complete a three-week randomized control study that includes: (1) an ecological momentary assessment, involving daily surveys delivered via phone four to five times per day to record daily sleep behaviors (time in/out of bed, napping), cannabis use (time, method, product), and motivation for cannabis use; (2) continuous actigraphy to objectively measure sleep; and (3) two 4-night in-laboratory sleep protocols to assess systematic responses to both sleep restriction and sleep extension, utilizing a randomized, counterbalanced, crossover design. Laboratory measurements will include sleep quality, quantity and dynamics (via full polysomnography; PSG), circulating cannabinoids before and after sleep (via venous blood), and cortisol (via saliva). This mixed-method approach—combining ambulatory assessments, observations of naturalistic cannabis use, and intensive laboratory measures with a controlled sleep protocol—will allow us to systematically determine the prospective relationship between sleep and cannabis use both within and between participants. This design will allow us to examine the bidirectional relationship between cannabis use and sleep (aim 1), as well as the relationship between circulating cannabinoids and sleep (aim 2). The rapid legalization of cannabis over the last decade has raised concerns about its potential negative impacts on health and safety, which are themselves influenced by sleep. Current research significantly lags behind consumers’ naturalistic cannabis use habits (e.g., the use of concentrates, which have stronger effects compared to smoked cannabis flower). The proposed study aims to address this gap.

NIH Research Projects · FY 2025 · 2025-06

Summary/Abstract The neurological, Down syndrome diagnosis is associated with various disease phenotypes that include cardiovascular, haematological and immunological disease processes.The increase in DNA copy number in Down syndrome, as a result of Trisomy 21, has led to the DNA dosage hypothesis, which posits that the level of gene expression is proportional to the gene's DNA copy number. The study of rare cases of partial Trisomy 21 has led to the identification of the dosage sensitive “Down Syndrome Critical Region”. Down syndrome individuals also experience cognitive decline in a syndrome that resembles Alzheimer's disease with its onset one to two decades earlier than typical Alzheimer's disease. The underlying hypothesis of this proposal is that epigenetic regulation at dosage sensitive genes on chromosome 21 contribute to the disease phenotypes associated with both Down syndrome and the co-occurring Alzheimer's disease. This hypothesis stems from our recent identification of >200 “Inactivation/Stability Centers” located on human autosomes. Inactivation/Stability Centers are ~1 megabase in size, are characterized by a novel epigenetic program that results in stochastic allelic inactivation of both protein coding and non-coding RNA genes. Inactivation/Stability Centers are also characterized by variability in allelic replication timing, where each allele can display either early or late replication timing. Two of the recently mapped Inactivation/Stability Centers are located on chromosome 21: one of them contains the Down Syndrome Critical Region, and the other contains the dosage sensitive, autosomal dominant, Alzheimer's disease gene APP (Amyloid Precursor Protein). This proposal is designed to elucidate the epigenetic mechanisms that are responsible for the allelic inactivation of the protein coding genes within the Down Syndrome Critical Region as well as at the APP gene. We have also found that the Down Syndrome Critical Region and the APP gene are syntenic regions in the mouse genome that are also within Inactivation/Stability Centers. This proposal is designed to use human and mouse cell-based assays to elucidate the molecular and cellular mechanisms associated with stochastic allelic inactivation of these dosage sensitive genes, and to determine the cellular phenotypes associated with the different expression states that are generated by stochastic allelic inactivation at these two critical locations on chromosome 21. This proposal challenges the existing models for autosomal random monoallelic expression, and if successful will establish a new paradigm for Down syndrome and the co-occurring early onset Alzheimer's disease that would include allelic inactivation of dosage sensitive genes in the generation of the cellular and disease phenotypes in patients with trisomy 21.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY Iatrogenic nerve injury represents one of the most feared surgical complications and remains a major cause of morbidity across all surgical specialties. Head and neck cancer surgery is a compelling clinical example of sig- nificant patient morbidity, where nerve damage provoked by surgical trauma is reported in up to 30% of patients, resulting in severe pain and increased stress exceeding that seen in many other cancer types due to the potential disfigurement. Surprisingly, no clinically approved technology currently exists to enhance intraoperative nerve identification, where techniques for nerve visualization still rely heavily on neuroanatomical knowledge and tra- ditional white light visualization. Fluorescence Guided Surgery (FGS) has made significant advances using FDA- approved near-infrared (NIR, 650-900 nm) fluorophores and cancer-targeted probes. However, clinical nerve FGS lags behind in the development and translation of cancer-specific contrast agents due to challenges in creating clinically relevant nerve-specific fluorophores. Importantly, although NIR fluorophores offer better tissue penetration and reduced light scattering compared to the visible spectrum, their utility is still limited in imaging depths and resolution for buried fine tissue structures (e.g., facial nerves, cranial nerves). Given the varied sizes and depths at which facial nerves are buried, and the critical need to minimize nerve damage, there is a com- pelling case for exploring beyond the NIR spectrum. The short-wave infrared (SWIR, 900–1700 nm) region emerges as a superior alternative for in vivo fluorescence imaging, with advantages including further reduced light scattering and increased tissue penetration, leading to dramatically improved resolution. These properties are particularly advantageous for the visualization and identification of delicate facial nerve tissues during head and neck cancer surgeries, where precise identification at depth can significantly mitigate the risk of iatrogenic nerve injury. However, the development of a SWIR nerve-specific small molecule has been a significant chal- lenge, because these fluorophores need to have a low enough molecular weight to cross the tight blood-nerve barrier junction, with a sufficient degree of conjugation for SWIR excitation and emission, inherently increasing their molecular weight. Our preliminary work has led to the development of over 400 novel oxazine-based small molecule fluorophores, providing quantitative structure-activity relationship (QSAR) modeling on the oxazine scaffold for nerve specificity management. Building on this foundation, we propose to synthetically develop first- in-class SWIR nerve-specific fluorophores that will push the boundaries of current imaging capabilities. The newly designed SWIR fluorophores will undergo rigorous validation for nerve specificity in rodent models, and the most promising candidates will be validated for their efficacy in buried facial nerve visualization during head and neck cancer surgeries in more complex cancer models. This work is poised to set a new standard for nerve preservation in one of the most challenging areas of oncologic surgery.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY The Prevotella genus is one of the most abundant and diverse genera in the oral cavity, and includes numerous prominent oral microbiome species that have been recognized for decades to play critical roles in mucosal health and disease. Accordingly, recent culture-independent studies have found oral Prevotella species to be strongly associated with a broad range of both oral and extraoral conditions, especially polymicrobial abscesses and multiple types of malignancies. Despite this, shockingly little is known about the biology of all Prevotella species, largely as a consequence of the notorious genetic intractability within the entire genus. Recently, we encountered this same limitation due to our ongoing studies of host interactions with our Prevotella human odontogenic abscess clinical isolates. While we have identified a variety of intriguing host interaction phenotypes, further mechanistic studies of these phenomena have been stymied by the lack of options for Prevotella genetic manipulation. Consequently, we have recently focused our attention on genetic system development for oral prevotellae. In our preliminary studies, we provide evidence of natural competence from 20 fully sequenced abscess clinical isolates of P. melaninogenica, and we have been able to exploit this ability to genetically manipulate multiple strains using cloning-independent methodologies. Furthermore, preliminary evidence suggests that the P. melaninogenica DNA uptake apparatus is likely to represent an entirely new class of natural competence machinery. Thus, we hypothesize that P. melaninogenica natural competence can serve as a model to break the genetic tractability barrier within the Prevotella genus and reveal the genetic basis for its natural competence ability. The goals of this project will be achieved in two Specific Aims that: 1) characterize the natural competence ability of oral Prevotella and 2) develop a complete genetic toolbox for detailed genetic studies of oral Prevotella.

NIH Research Projects · FY 2023 · 2025-06

PROJECT SUMMARY/ABSTRACT Our immune system is crucial for recognizing and suppressing cancers in the body. Unfortunately, melanomas, one of the most lethal skin cancers, can interact with and inactivate immune cells. Among the most effective anti-melanoma therapies are immunotherapies that reactivate or “train” the anti-tumor activities of immune cells. However, the effectiveness of immunotherapies is currently limited to ~30% of patients. Although the underlying causes are unclear, lack of responsiveness in patients is associated with insufficient infiltration of tumors by immune cells. Thus, studies aimed at elucidating melanoma:immune interactions and increasing the immune infiltration of tumors are required to improve immunotherapies. We discovered a potential way to increase the efficacy of immunotherapies by boosting infiltration of melanomas with tumor-suppressing immune cells using the plant sugar L-fucose. In a process called fucosylation, cells used L-fucose to modify proteins, affecting their maturation/function. We found that fucosylation is generally reduced during melanoma progression in humans, prompting us to test if increasing L- fucose/fucosylation levels in melanomas elicits therapeutically beneficial effects. Simply feeding L-fucose to melanoma-bearing mice reduces tumor growth and metastasis by >50% (Lau et al. Sci Signal 2015). Intriguingly, those smaller tumors contain 10-50 times more tumor-infiltrating lymphocytes (“TIL”) than tumors from mice not fed L-fucose. Genetically increasing the fucosylation of melanoma cells elicits the same effects, suggesting that melanoma fucosylation triggers anti-tumor immunity. We determined that CD4+/CD25- T cells are crucial for L-fucose-triggered recruitment of TILs including CD8+ T, NK, and DCs that suppress tumor growth. We identified the immune-regulating protein HLA-DRB1 as fucosylated, and its expression is crucial for TIL recruitment/tumor suppression, prompting our hypothesis that fucosylation of HLA-DRB1 triggers CD4+/CD25- T cell-mediated TIL recruitment and suppression of melanoma. However, how fucosylation regulates HLA-DRB1 to mediate anti-melanoma immunity, if those effects are due to increased tumor immunogenicity, CD4+/CD25- T cell function, or both, and if L-fucose/fucosylation can enhance immunotherapy efficacy or have prognostic utility is not known. We propose 3 Specific Aims (SAs) to test our hypothesis and address these questions: ·SA1: Determine how fucosylation regulates the localization and immune function of HLA-DRB1 ·SA2: Determine how systemic fucosylation affects CD4+/CD25- T cell biology. ·SA3: Determine if L-fucose/fucosylation enhances anti-PD1/TIL therapy and predicts patient prognosis Our goal is to provide key biological/mechanistic insights into melanoma:immune interactions, which will establish a basis for developing enhanced, fucosylation-based patient stratification and treatment strategies.

NIH Research Projects · FY 2025 · 2025-06

Adenosine to inosine (A-to-I) editing is a post-transcriptional modification which is prevalent in the developing nervous system. This chemical modification is catalyzed by adenosine deaminases (ADARs) acting on double- stranded RNA. Because inosine is structurally similar to guanine, A-to-I editing leads to inosine base pairing with cytosine and can change the encoded amino acid. A-to-I modification of many key neuronal mRNAs changes properties of their encoded proteins, including ion channels and mediators of synaptic transmission, signal transduction, and intracellular trafficking. Typically, editing increases as the brain matures during embryonic development, with less editing found in stem and neuronal progenitors and more editing in mature neurons. A- to-I editing is essential for normal brain development and abnormal editing has been associated with a number of diseases, including epilepsy, amyotrophic lateral sclerosis, psychiatric disorders, developmental disorders, and encephalopathy. The development of next generation sequencing approaches has enabled identification of A-to-I editing sites in different tissues. These approaches have identified specific neuronal target transcripts and the extent to which they are edited (editing efficiency varies widely and depends on transcript identity). Most mRNA target edit sites and their overall editing frequency are known. However, whether editing efficiency varies in different types of neurons within tissues and what fraction of a given transcript is edited in single neurons is not known. Recent developments in transcriptomic analysis of single cells allows assigning of individual neurons to various subtypes. However, common single cell RNA-sequencing (scRNA-seq) platforms, like 10x Chromium, use high-throughput short-read sequencing (e.g. Illumina Sequencing) to preferentially sequence the 3’UTR portion of transcripts to assign gene identity. As such, this approach does not allow comprehensive identification of editing events in coding regions of transcripts. On the other hand, approaches like Iso-Seq from Pacific Biosciences (PacBio), enable highly accurate, long-read sequencing (up to 15 kb), and do cover full transcripts in scRNA libraries. However, Iso-Seq read depth is too low (mostly due to sequencing costs) to sample enough transcripts per cell to assign detailed molecular identities. The goal of this proposal is to develop a methodology allowing for the simultaneous analysis of A-to-I editing sites in targeted RNAs (via long-read sequencing) in specific neuronal cell subtypes (characterized in detail via short-read sequencing). We will then benchmark our approach by performing a bulk RNA-seq in the same neuronal subtypes. Because multiple studies implicate A- to-I editing alterations in neurological and neuropsychiatric disorders, classification of A-to-I editing on a single cell level will enable a better understanding of disease pathogenesis and better targeting of potential treatments.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY This project is focused on understanding the mechanisms underlying brain vascular endothelial cell (BVEC) disruption following vascular brain injury (VBI). Vascular contributions to cognitive impairment and dementia (VCID) are the second leading cause of dementia, caused in part by VBI and blood-brain barrier dysfunction. VBI can cause the degeneration of BVECs, leading to brain capillary leakages that promote neuroinflammation and neurodegenerative disease. Accumulating evidence supports a role for ferroptosis, a form of programmed cell death involving iron overload and reactive oxygen species (ROS)-dependent lipid peroxide accumulation, in the degeneration of BVECs. A recent study suggested that ferroptosis can be inhibited following traumatic brain injury by the glycosaminoglycan hyaluronan (HA). We find that BVECs express elevated levels of the Cell Migration and Hyaluronan Binding Protein (CEMIP), a hyaluronidase, in patients with VBI. Elevated CEMIP expression is accompanied by increased HA synthesis and expression of the CD44 transmembrane HA receptor. Based on these findings, our overarching hypothesis is that HA protects BVECs from ferroptosis through a CD44-depedent mechanism. However, if CEMIP remains elevated, HA is digested, leading to the initiation of BVEC ferroptosis and BBB breakdown. We will test this hypothesis in the following specific aims: (1) To test the hypothesis that HMW HA inhibits BVEC cell ferroptosis through a CD44-dependent mechanism; (2) To test the hypothesis that HA digestion by CEMIP increases BVEC ferroptosis; and (3) To test the hypothesis that CEMIP inhibitors can prevent BVEC ferroptosis. These studies will provide mechanistic insights into the pathogenesis of VCID and will determine the feasibility of targeting HA signaling and CEMIP to protect BVECs, and possibly prevent blood-brain barrier dysfunction, following VBI.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY / ABSTRACT The Health Care Systems Research Network (HCSRN)-Older Americans Independence Centers (OAICs) AGING (Advancing Geriatrics Infrastructure and Network Growth) Initiative (R24AG045050/R33AG0577806) was launched in 2014 to address knowledge gaps around multiple chronic conditions (MCCs) in older adults by fostering an interdisciplinary research agenda. Our mission has been to counter the traditional single-condition, siloed approach to the care of older adults and advance the science of MCCs. The AGING Initiative is a highly productive, collaborative, transdisciplinary endeavor involving scientists from 20 HCSRN research centers, embedded within healthcare delivery systems caring for over 2 million persons aged 65 and older, in partnership with investigators from 15 premier, university-based centers established by the National Institute on Aging (the OAICs). In this competitive renewal, our overarching aim is to build on our successful efforts and collaborations, while taking the AGING Initiative in new, more ambitious, and sustainable directions. We will seek to promote health equity, further integrate diverse patients and care partners into our scientific activities, expand efforts to foster the careers of underrepresented minority (URM) scientists, and expand our efforts to engage with a more diverse group of patients/care partners and a broader array of organizations to improve the care of older adults with MCCs. Our specific aims are: (1) to foster research innovation and translation of research findings into practice through an MCCs Research Innovation, Translation, and Dissemination core that will: (a) fund targeted, interdisciplinary pilot grants (MCCs Innovation Grants); (b) promote inclusion of underrepresented populations in clinical trials relevant to the care of older adults with MCCs; (c) disseminate key research findings relevant to the care of older adults with MCCs; and (d) enhance engagement and collaboration with patients and care partners in research activities; (2) to foster the development of innovative methods under a Methods, Measures, and Data Core that will: (a) stimulate development and implementation of innovative research designs; (b) disseminate measures that “matter most” to patients, care partners, and healthcare providers; (c) identify new data sources and research partnerships; and (d) provide guidance and mentorship to junior investigators starting their “first” project; and (3) to foster the career development and success of new and early-stage investigators through an expanded nation-wide cohort of MCCs Scholars by: (a) enhancing mentoring and networking opportunities in direct response to feedback from current and former MCCs Scholars; (b) expanding efforts to identify and recruit URM junior investigators; (c) increasing collaborations with other NIA-sponsored research networks that have scholar programs to increase discourse on MCCs; and (d) building upon a highly successful collaboration between the American Geriatrics Society and the AGING Initiative (R25AG071488) to create and widely disseminate a new MCCs research curriculum.

NIH Research Projects · FY 2026 · 2025-06

Project Summary Developing brains over-wire and create excess synaptic connections. The nervous system then undergoes neuronal remodeling by implementing structural changes through pruning of superfluous neurons and synapses. Neurons and glia communicate intensively during the remodeling process, but the mechanistic basis of neuronal or glial signaling remain understudied and poorly defined. Disrupted remodeling has implications on behavior and circuitry in neurodevelopmental disorders, like autism spectrum disorder (ASD), and even neurodegenerative diseases like Alzheimer’s, highlighting the importance of defining this process. There are strong correlations between broad changes in neural circuit connectivity and behavioral changes in ASD animal models and patients. This has led to the proposal that broad changes in neuronal remodeling during development cause behavioral changes, but this idea has not been tested directly and the link between structural changes in circuits (e.g. local pruning of neurites) and behavior remains unclear. Drosophila Pair1 neurons, a circuit where 6 neurons drive reversal behavior in the larva and adult, provide an excellent model to study both neuronal and glial molecular mechanisms of remodeling, and investigate how perturbation of remodeling affects behavior and circuit morphology. In preliminary work I have defined the morphology of their local pruning and identified caspase signaling as a key neuron-intrinsic pathway that drives pruning. In Aim 1, I will investigate intrinsic mechanisms for Pair1 local pruning by exploring known caspase cascades in the context of Pair1 pruning using loss-of-function studies and caspase indicators with live ex-vivo imaging. In Aim 2, I will determine if astrocytes are responsible for activating Pair1 remodeling and clearing pruned debris, and conduct a targeted genetic screen for glial molecules required for pruning of Pair1 neurons. In Aim 3, I will block intrinsic remodeling cues in Pair1 neurons to ask if altering Pair1 local pruning affects assembly of adult circuitry and behavior. Together, these aims will contribute new fundamental knowledge to the remodeling field, which ultimately will help us understand how brain circuits are optimized and how changes in remodeling might affect behavioral phenotypes in neurological disorders and disease. The training potential for me is very high in this project, and my proposed research will help me achieve both my research and professional training goals. I will deeply expand my knowledge in both the glial and neurodevelopmental biology fields, while also gaining many new technical skills in the laboratory. Outside of the laboratory, I will consistently seek career development opportunities to advance my communication skills, leadership, and equity and inclusion education and impact. I am strongly supported in my training goals from my sponsor Dr. Marc Freeman, the Vollum Institute, and OHSU and know I will succeed with the support of this award.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY The goal of our work is to uncover the cells and pathways that affect mucus changes downstream of hormonal changes, with the long-term goal of using this information to inform the development of novel, non-hormonal therapeutics targeting the endocervix that would inhibit (contraception) or enhance fertility. Endocervical cells produce mucus, and changes in mucus consistency during the normal menstrual cycle influence human health and fertility. For natural fertility to occur, mucus must change from a highly viscous form that blocks sperm penetration to a highly fluid form that allows for sperm penetration and fertilization. Mucus also blocks entry of pathogens in the gravid and non-gravid uterus, playing a key role in overall health of the uterus and upper reproductive tract. Despite the known biological and clinical importance of the endocervix and mucus in fertility, contraceptive studies in the endocervix have been methodologically limited as clinical studies of the endocervix lack discriminating, biologically meaningful endpoints and laboratory studies are limited by availability of cellular and animal models. Meanwhile, research in the airway and other non-reproductive organs have made major advances in both understanding the physiology of mucus properties and development of research tools for investigating epithelial-mucus interactions. Airway studies have revealed that mucin glycoproteins and regulation of their hydration via epithelial ion channels underpin the biophysical and rheological properties of secreted mucus. Our study builds on our prior work establishing that the endocervical epithelia is similar to epithelia from other mucosal organs, including the mucin protein constituents, expression and function of ion channels, and testability of cellular mucus changes. Applying the most recent knowledge of epithelial mucus biology to cycle-dependent fluctuations of endocervical mucus characteristics leads us to hypothesize that female sex hormones estradiol and progesterone regulate mucus biophysical properties, mediated by alteration of epithelial cell populations and states, mucus protein composition, and ion channel transporter activities in the endocervix. To do this, we will leverage a unique multi-disciplinary, multi-institutional collaboration bringing together expertise in reproductive health and the endocervix, epithelial cell biology, mucus and mucin biophysical properties, ion channel biology, and next-generation sequencing studies and bioinformatics in the reproductive tract. We will combine 1) an established clinical trial model of steroid hormone ablation and replacement to collect endocervical cells and mucus under highly controlled hormonal conditions in humans with 2) a novel in vitro model of conditionally reprogrammed endocervical cells that leverages an array of phenotypic endpoints of mucus changes adapted from highly successful airway drug discovery programs. By combining clinical and in vitro studies, we expect to discover and validate novel mechanisms responsible for regulation of mucus biophysical properties during the menstrual cycle in the cervix and in turn provide a scientific basis for clinical implication of mucus as a tool to control fertility.