North Carolina State University Raleigh

NIH Research Projects · FY 2026 · 2026-06

Summary. Aptamers – a versatile class of synthetic, nucleic-acid-based receptors – offer unique advantages for molecular sensing. Because of the hydrophilicity of oligonucleotides, for example, aptamers offer a key advantage relative to antibodies in terms of remaining soluble even when partially or fully unfolded. This distinctive characteristic renders it straightforward to engineer aptamers that fold and unfold in response to target binding and dissociation. In turn, this coupling of molecular recognition to a large-scale conformational change enables the construction of biosensors supporting real-time in vivo molecular measurements and chemo- responsive, materials whose properties change in response to a specific molecular cue. A major disadvantage of aptamers, however, is that their affinity and specificity are often insufficient for use under physiological conditions. Here, we present strong evidence that this poor performance is not an intrinsic limitation of nucleic acids, but instead arises from the limitations of conventional aptamer selection techniques. Building on this, we describe advanced aptamer selection and maturation approaches that we believe will surmount these limitations. In Aim 1, we propose the development of a powerful new aptamer selection approach, nuclease-assisted systematic evolution of ligands by exponential enrichment (NA-SELEX), which eliminates the need to attach either the target or the library to a solid support. This provides many important advantages over existing selection approaches, including the ability to simultaneously apply both thermodynamic and kinetic stringency and to easily carry out aptamer selection under physiologically relevant conditions, leading to enhanced in vivo aptamer performance. In Aim 2, we will complement NA-SELEX by developing an innovative “aptamer maturation” approach, Motif-SELEX, that can greatly improve the performance of underperforming first-generation aptamers. Motif-SELEX does this by providing new opportunities to explore the deep sequence space for a given aptamer during the selection process in order to optimize the geometry and complexity of that aptamer’s ligand-binding site, thereby increasing both its affinity and specificity. Finally, in Aim 3, we will showcase the improved performance and utility of the resulting aptamers by converting them into real-time biosensors to enable the currently unachievable feat of measuring a chemically diverse set of biomedically-important, low-concentration small-molecule targets in vivo. In the short term, the successful conclusion of our research program will yield sensors that enable clinically and scientifically useful real-time in vivo measurement of numerous biomedically- important low-molecular-weight analytes. The more far-reaching impact of this work will be the development of methods supporting the selection of aptamers with unprecedentedly high performance, greatly expanding the utility of these reagents for a wide range of biomedical applications.

NIH Research Projects · FY 2026 · 2026-06

Abstract While much of the human microbiome research is focused on bacteria, one of the most prevalent yet underappreciated components of the human microbiome is the methanogenic archaea (methanogens). They catalyze the unique methanogenesis metabolism responsible for methane production in the human body. About 1/3 of the healthy adults are breath methane positive, and at least 2/3 are tested positive for fecal methanogens. Although growing evidence has suggested both beneficial and detrimental roles of methanogens in digestive health, the foundational biology supporting these roles remains elusive. Our working hypothesis is that methanogens of the Archaea domain form interdomain syntrophic partnerships (syntrophy) with their surrounding microbiome of the Bacteria domain, specifically defined as methanogen-bacterial syntrophy (MBS) in this application. Normally, MBS is an energy-efficient partnership that benefits food digestion and thus contributes to homeostasis. However, when MBS becomes dysfunctional for reasons yet to be known, growth of methanogens becomes dysregulated, leading to dysbiosis marked by enrichment or depletion of methanogens. The former is already defined by the American College of Gastroenterology as Intestinal Methanogen Overgrowth (IMO) which is strongly associated with constipation suffered by millions of Americans, particularly those with irritable bowel syndrome (IBS). We hypothesize that a shift in redox and dietary environment causes dysfunction in MBS, because 1) methanogens are strict anaerobes that are very sensitive to oxygen, rapidly lose activities in a matter of hours, and require a highly reduced environment for robust growth, and 2) gut microbiome and their functions vary greatly depending on the diet. That is, when the gut microbiome is challenged with oxidative stress, oxidative dysfunction occurs in MBS which leads to depletion of methanogens. When facing reductive stress, reductive dysfunction develops in MBS which contributes to enrichment of methanogens. In terms of diet, we hypothesize that there are multiple yet-to-be-determined subtypes of MBS that drive digestion of glycan, fat, and protein, respectively. Depending on the specific MBS subtype and diet a person has, either enrichment or depletion of methanogens occurs. At least four major technical and knowledge gaps exist in our working hypothesis: 1) What is the technical approach that models both oxidative and reductive stress for microbiome studies? 2) What are the MBS subtypes that drive digestion of various organic matters? 3) What are the potential molecular mechanisms that mediate MBS under different redox and nutrient conditions? 4) What is the impact of MBS on digestive health in vivo? To fill these gaps in this application, PI Lyu (PhD microbiologist) and Co-I Pimentel (MD physician scientist) seek to leverage our Anaerobic Tandem Chamber System (ATCS) platform to model redox stress, combine both cultivation- dependent and -independent approaches to determine MBS subtypes, build in vitro co-culture models for MBS, and test the impact of MBS on digestive health in rats in vivo for IBS-like symptoms. These efforts will help us achieve our shared long-term goal of understanding the role of methanogens in human health. The contribution of the proposed research is significant as it seeks to move away from documenting correlations and move toward investigating functions using live microbes, a necessary step to tap the full potential of microbiome sciences. Another significance is that it aims to develop in vitro and in vivo models to include both archaea and bacteria under various redox and nutrient conditions. This will add more dimensions to digestive and microbiome research. The findings in this proposal will create fundamental knowledge about the biology of human-associated archaea, advance understanding of interdomain interactions between archaea and bacteria, and begin to unravel the interplay between microbiome, redox, and diet in the context of IMO and IBS. This will ultimately benefit efforts in developing microbiome-, redox-, and diet-based diagnostics and therapeutics for IMO and IBS, helping us maximize the benefits and minimize the detriments of microbiome on digestive health.

NIH Research Projects · FY 2026 · 2026-06

Project Summary Neurodevelopmental disorders, including autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD), have dramatically increased in prevalence, suggesting that environmental factors contribute to their etiology. While exposure to environmental contaminants is increasingly linked to neurodevelopmental abnormalities, the underlying mechanisms remain poorly understood. Mast cells—immune cells present in nearly all tissues, including the brain—are uniquely positioned to bridge environmental exposures with neuroimmune regulation. Their ability to rapidly release bioactive mediators in response to immune challenges makes them critical players in chronic peripheral and central inflammatory responses. However, their role in neurodevelopmental disorders remains largely unexplored. Our previous findings show that developmental exposure to Firemaster® 550 (FM550), a widely used flame retardant associated with neurodevelopmental impairments, induces persistent mast cell hyperactivity. This hyperactivity amplifies sickness and neuroinflammatory responses to immune challenges, leading to behavioral deficits in a sex-specific manner. We hypothesize that FM550 exposure reprograms mast cell progenitors into a hyperactive state, resulting in tissue- specific immune dysregulation that increases susceptibility to systemic inflammation and neurodevelopmental disorders. This project will address this hypothesis through three independent yet interconnected aims: (1) determine the distinct contributions of peripheral and brain mast cells to systemic and behavioral outcomes resulting from developmental FM550 exposure, (2) assess whether FM550 exposure induces tissue-specific mast cell hyperactivity, with peripheral mast cells upregulating mediators that drive acute sickness and systemic inflammation while brain mast cells promote microglial activation and blood-brain barrier disruption, and (3) determine whether FM550 exposure induces epigenetic modifications that enhance signaling in key mast cell activation pathways, revealing how environmental toxicants shape long-term neuroimmune susceptibility.This interdisciplinary project integrates behavioral neuroscience, immunology, and toxicology with advanced molecular and epigenetic techniques. By elucidating the mast cell-mediated mechanisms linking environmental toxicants to neurodevelopmental disorders, this research will provide critical insights into environmentally driven neuropsychiatric conditions and generate a comprehensive dataset that could be used to identify new targets for intervention.

NIH Research Projects · FY 2026 · 2026-05

Project Abstract Many solid cancers are inoperable due to either tumor size or because the tumor is attached to, or near, major blood vessels, vital organs or other critical tissues. Focal ablation modalities utilizing a variety of energy forms to destroy/debulk tumor tissues are frequently used in the management of inoperable solid cancers. More than 45,000 tumor ablation procedures are performed each year. Three thermal ablation techniques, radiofrequency ablation (RFA), cryoablation (CA) and microwave ablation (MWA), account for more than 80% of all procedures, but residual tumor deposits often lead to recurrence rates that are 2 to 10 times higher than recurrence rates following surgical resection. As a result, patients with inoperable tumors have worse survival outcomes compared to patients with resectable tumors. Integrated Nanosecond Pulse Irreversible Electroporation (INSPIRE) is a novel, minimally invasive solid tumor ablation modality. This thermally-regulated approach uses ultrashort alternating polarity electrical pulses to destabilize tumor cell membranes while preserving the integrity of nearby vital structures. A key advantage of INSPIRE, which forms the crux of this proposal is the flexibility in energy delivery and numerous parameters combinations that allows us to modulate cell death mechanisms to favor immunogenic pathways, and subsequent antitumor immune responses. Induction of robust antitumor immunity following focal ablation is crucial for eliminating residual tumor cells and preventing local or distant recurrence which can be enhanced with secondary immunotherapy agents such as checkpoint inhibitors. Based on our preliminary studies, we hypothesize that there is a ‘best’ or most immunogenic INSPIRE protocol which maximizes the resultant antitumor immune. We further hypothesize that an adjunctive immunotherapy can boost the INSPIRE-induced antitumor response, thus enhancing both local and systemic tumor control. These hypotheses will be tested in an aggressive, transplantable murine melanoma model before translation into comparative oncology trials in pet dogs with spontaneous melanomas via three aims: Aim 1: Develop and validate INSPIRE protocols to maximize tumor-specific immunity; Aim 2: Evaluate the timing of adjunctive immunotherapy relative to INSPIRE; Aim 3: Validation of Combinatorial INSPIRE in a Spontaneous Large Animal Model of Disease. These aims will determine the treatment parameters which maximize immune stimulation via INSPIRE, improve systemic anti- tumor immune responses via optimized adjunctive immunotherapy regimens, and demonstrate clinical superiority of this combined approach in a relevant large animal model. Our long term goal is to develop a novel, minimally invasive focal treatment paradigm that is capable of preventing recurrences and eliminating metastatic deposits. Successful completion of the proposed project will support further translation of INSPIRE plus adjunctive immunotherapy into human trials while providing a state-of-the-art treatment for thousands of companion animals per year.

NIH Research Projects · FY 2026 · 2026-05

Project Summary: Over 300 million individuals are affected by asthma worldwide, and allergic asthma, the most common form of asthma, affects more than 50% of asthmatic adults and 80% of asthmatic children. The elevated levels of proinflammatory mediators contribute to the initiation and progression of pathology in allergic lungs. Therefore, elucidating the cellular and molecular mechanisms of expression of proinflammatory mediators in allergens-induced inflammation is highly significant in terms of formulating anti-inflammation therapeutic strategies. These therapies primarily aim at the mitigation of granulocytic recruitment and pro-inflammatory signaling responses. Therefore, harnessing the capabilities of endogenous regulators that mitigate proinflammatory responses is a viable therapeutic option. The overall objective of this proposal is to delineate the cell-specific role of Tristetraprolin (TTP) in allergens-induced respiratory inflammation and to explore the potential of modulation of TTP expression in the mitigation of allergic respiratory inflammation. Our published and preliminary data suggest that TTP is an endogenous regulator of active inflammation in diverse inflammatory conditions. TTP is known to be expressed in a wide variety of cells including epithelial cells and immune cells where it regulates the expression of cell-specific target mRNAs. However, the cell-specific role of TTP in allergens-induced respiratory inflammation and the beneficial effects of modulation of TTP expression in allergens-induced respiratory inflammation remain unexplored. Accordingly, in this application, our central hypothesis is that TTP mitigates allergens-induced respiratory inflammation. The specific aims include, 1) Using cell- and lineage-specific TTP deletion mice, we will test the hypothesis that TTP modulates allergens-induced respiratory inflammation, 2) Test the hypothesis that pharmacological stabilization of TTP mitigates allergens- induced respiratory inflammation. In this aim, we will explore the effect of TTP overexpression or TTP stabilization in allergens-induced chronic respiratory inflammation. These studies will have a transformative impact on our mechanistic understanding of the role of TTP in the pathophysiology of allergens-induced environmental respiratory disease that may lead to the development of cell-specific therapeutics towards allergens-induced as well as other inflammatory diseases.

NIH Research Projects · FY 2026 · 2026-03

Abstract Functional development of the body plan and organs requires tissues to adopt the specific three- dimensional morphologies required for function. Defects in tissue shape lead to functional deficits with impacts ranging from reduced or absent organ function to peri-natal lethality. The complex tissue shapes required for function are built from simple precursors by the spatially and temporally coordinated activity of individual cells. In many cases, final tissue shape is driven exclusively, or nearly exclusively by a single class of cell dynamic, such as cell shape remodeling, rearrangement of cells with respect to one another, or localized or oriented cell divisions. However, many tissues rely on a combination of multiple behaviors to build complex morphologies, which presents a challenge as these behaviors are incompatible within individual cells. Thus, a major challenge in human health and developmental biology is to understand how multiple classes of cell dynamics occurring concurrently in the tissue are successfully integrated despite their mutual antagonism in individual cells. A key related question is how these disparate behaviors are controlled and spatially patterned within tissues, as this spatial control is key to successful integration. In this proposal, we will use the mouse cranial neural plate as a model system for understanding how behaviors are integrated. This is an attractive system, as at least three mutually disruptive behaviors occur: apical constriction, planar polarized cell intercalation, and oriented cell divisions. We will use a combination of mouse genetics, bespoke cellular and subcellular live imaging approaches, quantitative in toto imaging of tissue organization, and a combination of molecular and transcriptomic approaches. Together, these experiments will directly define the role of each class of cell behavior in generating final tissue shape, elucidate the spatially delimited developmental signaling systems that control individual cell behaviors, and determine how mutually disruptive behaviors are integrated to drive complex tissue shape outcomes.

NIH Research Projects · FY 2026 · 2026-01

Project Summary Caenorhabditis elegans, a well-studied nematode, is an important model to study the genetic mechanisms that regulate aging, longevity, and health. Most aging studies in this model organism measure the survival of a population through time, and factors that extend or shorten lifespan are observed by a shift of these survival curves. An important limitation of this type of analysis, is that individual animals cannot be tracked, and thus the identity of each individual is lost in each day where survival is scored. Tracing the aging trajectories of individual animals within a population is necessary to better understand how early life events affect the rate of aging and longevity. In addition, populations tend to show very variable lifespans even though they are genetically identical, and are exposed to the exact same conditions. Better understanding this variability and how different interventions can modulate longevity differently in different animals can only be possible if animal identity can be detected. Here, we propose to develop a new system, BOWiD, which provides a readable and stable barcode for each animal in a population, and thus allows extracting the identity of each worm. Barcodes will be imprinted in animals by turning on a fluorescent protein in a subset of cells. Reporters will be activated randomly by the Cre-lox recombination system. To accomplish this, we will focus on: 1) Developing robust components needed for BOWiD, including inducible promoters for Cre, identify robust cellular landmarks and labeling approaches, and developing machine learning pipelines to read barcodes, 2) Developing and implementing BOWiD for animals cultured on traditional agar plates, and analyze aging trajectories, and 3) Developing and implementing BOWiD for animals cultured in microfluidic devices, where trajectories of the structure of aging neurons will be analyzed, in a high-resolution setup.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Since any antimicrobial drug use has the potential to increase resistance, a key goal is to align antimicrobial drug use (AU) with the prescribing guidelines in veterinary settings. Pet owners have expressed a preference for receiving antibiotics over watchful waiting including in cases where the benefit of antibiotics is unclear and a lack of awareness of the risk of antimicrobial use on their pets and the greater society have been found to be barriers to judicious use. Information is lacking on the impact of available educational resources to reduce dog owners' intentions to request antibiotics and increase their confidence in alternatives to antimicrobial drugs. Since 45.5% (59.8 million) households own an estimated total of 89.7 million dogs, and since up to 70% of dogs receive antibiotics for the treatment of acute diarrhea, it is crucial to test the impact of targeted resources designed to persuade dog owners that antimicrobials are not needed in these cases. To foster antimicrobial stewardship in veterinary settings using training or education tools, we will use the Theory of Planned Behavior to quantify the knowledge, attitude, subjective norm (perceived social pressure regarding guidelines), and perceived behavioral control (perceived ability to follow the guideline) related to requesting antimicrobials for dogs with a common medical condition (i.e., acute diarrhea). We will explore whether a short educational video on antimicrobial stewardship or a handout summarizing evidence-based guidance with accompanying citations can impact dog owners' opinions on antibiotics for canine acute diarrhea and define characteristics of antimicrobial use resources that dog owners identify as effective in changing their intention to request antimicrobials for canine acute diarrhea. The impact of these resources will be assessed using a cross-sectional survey of dog owners in the United States randomized to 3 arms (no resource, educational video, written summary of evidence-based guidance) followed by focus groups with participants drawn from each group to define which aspects of AS resources are effective in reducing dog owners' intent to request antimicrobial drugs and confidence in alternatives to antimicrobial drugs. The outcome of these aims will facilitate a greater understanding of the potential impact of AS resources on aligning dog owners' expectations for antibiotics with prescribing guidelines and inform further research into the creation and implementation of resources for dog owners to further the goals of antimicrobial stewardship.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Anterior cruciate ligament (ACL) injuries impact more than 250,000 people in the U.S. annually. The fastest rising injury rates are in children and adolescents with significant growth remaining. Reconstruction of a completely torn ACL is becoming increasingly popular to treat these injuries in children of all ages to restore knee stability and permit the return to sports while limiting secondary injuries to other structures. Failure rates after ACLR for pediatric patients are higher than adults, with similar risk for long-term joint degeneration. For complete injuries, a variety of techniques have been proposed, including epiphyseal, complete transphyseal, and partial transphyseal approaches. Yet, comparisons of long-term joint degeneration between techniques have not been performed. For partial ACL injuries, increasingly, surgeons will reconstruct the ACL; however, it is unclear if outcomes are better by replacing the ACL entirely or by preserving the functional bundle. Human pediatric joints for in-vitro cadaveric testing are extremely limited, and animal models are often skeletally mature. To better assess surgical techniques for skeletally immature patients, it is necessary to use a model that accounts for age- specific ACL function as well as match the biomechanical properties of the allograft to the native ACL. The porcine model is useful to study the ACL during growth and established the age-dependent nature of ACL bundle function. Thus, the objective of this proposal is to better understand outcomes after complete and partial ACL reconstruction during growth and use this knowledge for selection of appropriate surgical techniques. To accomplish this objective, we will leverage our multi-disciplinary experience to accomplish the following aims. In Aim 1, we will determine how ACLR technique influences the ability to restore joint stability and reduce joint degeneration at different stages of skeletal maturity. In Aim 2, we will determine how the location of partial ACL injury (anteromedial (AM) or posterolateral (PL)) influences the ability of partial or complete ACL reconstruction to restore joint stability and reduce joint degeneration. Successful completion of these aims will guide surgical technique section after complete and partial ACL injuries in children and adolescents.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract Asthma, a chronic respiratory disease, afflicts millions of people in the U.S. and exacts high financial burden on the healthcare system. As people spend most of their time indoors, allergic asthma and atopy are frequently induced by exposure to a variety of indoor biological allergens. Sensitization to cockroaches and chronic exposure to cockroach allergens are well recognized as important risk factors in the development and prevalence of asthma in children, especially in low-income urban and rural households. Exposure to bacterial and fungal toxins, such as endotoxins and glucans respectively, is also linked to pulmonary inflammation and exacerbation of asthma symptoms. A recent National Health and Nutrition Examination Survey study reported a strong positive correlation between the presence of cockroach allergens and endotoxins in dust sampled from the homes of asthmatic children. However, there is a conspicuous gap in our understanding of whether the microbes that cockroaches disseminate into the home environment re-shape the indoor microbiome and exposome, and consequently impact indoor environmental quality and health outcomes. Our long-term goal is to identify major cockroach-associated asthma triggers in the indoor environment, ascertain their impacts on the health of asthmatic children, and develop sustainable strategies to mitigate and abate their impacts. The goal of this exploratory study is to elucidate the causal relationships and linkage between the prevalence of cockroaches and several indoor pollutants of microbial origin. Preliminary results support the central hypothesis that perennial cockroach infestations degrade indoor environmental quality (IEQ) in high-risk low-income homes by producing not only a well-recognized cocktail of potent allergens, but also endotoxins, glucans, and mycotoxins that can exacerbate the impacts of the allergens as asthma triggers. The proposed approach includes two specific aims: 1) Determine the causal relationship between cockroach infestations and indoor microbial toxins , and 2) Determine the role of cockroaches in proliferation, vectoring, and sustaining microbes and toxins that are linked to respiratory disease and asthma . The project innovates by integrating in-home sampling with metagenomic, metabolomic, and proteomic approaches to identify novel asthma triggers that contribute to health disparities, and by using environmental interventions to quantify the effects of cockroach infestations on the prevalence of microbial toxins indoors. The impacts of this study include improving our understanding of the mechanisms by which cockroaches shape the indoor microbiome and identifying emerging biological pollutants that exacerbate respiratory health in low-income cockroach-infested homes. Future directions will explore 1) the roles and mechanisms of cockroach-associated microbial toxins in exacerbating the impact of allergens in a mouse model, and 2) the effects of targeted environmental interventions that eliminate cockroaches on reducing all cockroach-associated contaminants (allergens and microbial toxins) and improving health outcomes in asthmatic children.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Although lower limb (LL) prosthetics can restore basic mobility, not being able to handle different locomotion tasks has been the top complaint from patients about the traditional passive prosthetic legs. Recently, advanced powered prosthetic legs have become clinically available. They have been shown assisting various locomotion tasks, improving walking efficiency, lessening undue compensation from intact joints, and reducing secondary injuries (such as back pain). However, none of those benefits can be guaranteed until the control of powered LL prostheses is personalized appropriately. Current clinical practice in personalizing powered LL prostheses has been performed manually and heuristically. This approach is time and labor intensive. Due to time restrictions in clinical visits, prosthetists could only personalize a subset of prosthesis control parameters, limiting the device tuning precision and the user’s locomotion functions. In addition, manufacturers need to train prosthetists with specialized knowledge about control of a specific prosthesis prior to personalizing the device for a patient. Since the number of these specialists is limited, accessing their clinics is challenging and costly to LL amputees. Therefore, a new solution to transform the current clinical practice is urgently needed. To address this clinical need, our team has pioneered reinforcement learning (RL)-based approach that can simultaneously personalize high-dimension prosthesis control parameters automatically, quickly, and safely. Yet, how to translate this innovative method into clinics, and whether it benefits the prosthetists and amputee users have not been investigated. The objective is to investigate the efficacy of our RL method in prosthesis personalization clinics. We propose a RL-based Recommendation Interfacing System (RISE) that automatically recommends the prosthesis control parameters to prosthetists to meet their device tuning needs, which are based on their individual clinical judgment and the user’s verbal feedback. Our central hypothesis is that compared to current clinical practice, RISE will significantly improve the work efficiency of a prosthetist, i.e., they can perform personalization of any powered prosthesis quickly and accurately without a need to learn engineering control of powered prostheses. This project is clinically significant, because not only it improves the efficiency of clinical practice, but also it increases LL amputee users’ accessibility to powered prosthesis and its tuning clinics, improves their locomotion functions, and reduces the cost to amputees associated with the powered prosthesis use.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY Perfluoroalkyl and Polyfluoroalkyl Substances (PFAS) are synthetic compounds found in the environment and linked to adverse health outcomes. However, understanding the causal effects of PFAS mixtures on various health outcomes remains limited. This project addresses this critical knowledge gap by employing innovative spatial and high-dimensional causal inference methods to enhance our understanding of PFAS health effects. By bringing scientific rigor to observational studies, the project aims to overcome the challenges posed by spatial dependence, various sources of bias, and high dimensionality in PFAS studies. This project leverages two valuable health outcomes data sources relating PFAS in drinking water: (i) a nationwide cohort of Medicare beneficiary data and (ii) a North Carolina electronic health records cohort. These cohorts have been linked to PFAS concentrations in the ground water sources and in the tap water used for drinking. These data of health outcomes contain individual-level information and provide comprehensive resources for enhancing the understanding the effects of PFAS on population health. Based on previous epidemiological analysis of associations with PFAS exposure in the two cohorts we have identified three statistical challenges. First, spatially- dependent outcomes violate standard assumptions of causal inference, necessitating a rethinking of the potential outcomes framework. Second, various sources of bias are present such as preferential sampling and non- random actualization of PFAS exposures. Third, the high dimensionality of the PFAS chemical mixture and health responses introduces computational and stability issues, requiring new dimension-reduction methods. By developing a suite of methods tailored to address these challenges, this project marks a significant advancement in the field of epidemiological research. The proposed methods will enable accurate estimation of causal health effects in observational spatial studies, providing refined insights into the impact of PFAS mixtures on a range of health outcomes. To tackle these challenges, the project outlines three specific aims. Aim 1 focuses on developing spatial causal inference methods for preferentially-sampled data, accounting for bias in sample-location selection. Aim 2 devel- ops spatial causal inference methods for multiple outcomes and exposures that use tensor regression model to explain variation in the health effects by exposure, response, and spatial resolution. Aim 3 develops a structured statistical protocol for spatial causal inference analysis of multiple exposures and outcomes, with application to health effects of PFAS at the state and national levels. These aims not only address the specific challenges in PFAS studies but also have broader implications for epidemiological research dealing with spatial dependence and high-dimensional data. Through the development of these innovative methods, this project aims to transform the analysis of PFAS studies, contributing to advancements in the field and improving our understanding of the health effects of PFAS exposure.

NIH Research Projects · FY 2026 · 2025-06

Cytokinesis, the physical separation of a mother cell into two daughter cells, occurs by the constriction of a ring of actin filaments, myosin, and other evolutionarily conserved proteins. As cytokinesis is central to the propagation of species, the development of multicellular organisms and critical to the progression of fibrotic diseases and cancer, understanding the mechanisms that govern cytokinesis will broadly impact science and biomedical fields. How the contractile ring generates force remains unknown in part due to our lack of knowledge of its molecular organization, dynamics, and internal mechanics. Our recent progress suggests that the contractile ring produces tension by a mechanism similar to the molecular clutch mechanism of cell migration. According to this a molecular clutch mechanism, membrane-bound protein complexes called nodes engage with the dynamically contracting actin network and transmit the contractile forces to the plasma membrane/cell wall to drive cytokinesis. We will determine the mechanisms that govern force production during cytokinesis with an immediate emphasis on the forces created in the main bundle of actin filaments and how they are transmitted to the plasma membrane/cell wall. We are uniquely positioned to address these questions owing to our innovative combination of genetics, quantitative cell biology, biophysics, single molecule localization microscopy (SMLM), reconstitution assays and simulations. We divide our immediate plans into the different functional layers of the ring: the anchoring, the contractile, and the actin-microtubule crosstalk layers. I- Forces across the anchoring layer. We will determine how nodes respond to mechanical load by measuring their dynamics and molecular organization in constricting contractile rings with a combination of genetics, SMLM methods, tension measurements and NanoTrax. This work will reveal for the first time the mechanical impact of transmitting forces from the contractile actin network to the plasma membrane/cell wall through nodes. II- Forces within the contractile layer. We will determine the dynamic and molecular organization of the actin network in the contractile ring with a combination of genetics, quantitative microscopy, SMLM and biochemical methods. These results will provide the first measurements of actin dynamics and organization within a contractile ring and contribute critical information about the forces that drive constriction. III- Forces in the actin-microtubule crosstalk layer. We will determine how the physical interactions between microtubules and the contractile ring impact cytokinesis with a combination of genetics and quantitative microscopy methods. These results will provide critical knowledge in understanding how actin-microtubule crosstalk contributes to cytokinesis and nuclear positioning during mitosis in fission yeast. This program will determine the foundational mechanisms of tension production in fission yeast cells and will provide the essential steppingstones for our long-term goals to determine the evolutionarily conserved core mechanisms that drive force production and transmission in cytokinesis.

NIH Research Projects · FY 2026 · 2025-06

Project Summary Left-right (LR) differences in size, shape and/or anatomical position exist in almost every organ system. Consequently, abnormal LR asymmetry (known as heterotaxy, HTX) leads to a catastrophic syndrome of often-fatal birth defects. While the early embryonic events that establish global LR asymmetry have been well studied, it is the later-stage, organ-specific LR asymmetric morphogenesis events that determine normal anatomy; yet, for most organs, the molecular and cellular processes that sculpt their individual LR asymmetries have not been elucidated. This application addresses the asymmetric morphogenesis of the stomach, an organ whose familiar leftward curvature is an archetypical laterality among vertebrates. Our previous work has shown that the curvature of the stomach depends on LR asymmetric endoderm cell rearrangements, which cause preferential thinning and expansion of the left stomach wall. However, preliminary data suggest that this process alone is insufficient for proper curvature, and that events in the right stomach wall, as well as asymmetries in other tissue layers, including mesoderm and extracellular matrix (ECM), are also required for correct laterality. We hypothesize that stomach curvature is generated by interdependent LR asymmetries in endoderm rearrangement, mesoderm differentiation, and ECM composition. In the proposed project, we will integrate two powerful and complementary approaches to determine the individual and combined roles of different types of asymmetries in stomach curvature. First, we will use the exceptionally amenable frog embryo to execute tissue- and side-specific assays of gene function in vivo, along with innovative tests of tissue-mechanical properties. Second, we will use computational models of stomach morphogenesis to simulate multiple cell- and tissue-level asymmetries in silico, making testable predictions and revealing non-obvious, abiogenic effects on organ topology. In Aim 1, we will identify the LR asymmetries within each tissue layer that are required for stomach curvature in vivo. In Aim 2, we will employ computational modeling to determine the combinations of cell- and tissue-level LR asymmetries that are sufficient to reproduce realistic curvatures in silico. Aim 3 will utilize both in vivo and in silico modeling to ascertain the cell and tissue-level mechanisms underlying abnormal curvatures in HTX. Rigorously comparing computational simulations with anatomical data from nano-CT scans will ensure cross-validation and full integration of in vivo and in silico results throughout the project. Our findings will yield multilevel insights into the pathogenesis of LR defects in the stomach, with implications for the cell- and tissue-level events that underlie the emergence of more complex lateralities in other organs. This project will also highlight the value of in silico paradigms for morphogenesis research.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY/ABSTRACT NIA has a rich portfolio of “deeply phenotyped” small- to mid-sized longitudinal studies which are an underutilized resource of psychosocial, behavioral, and biomarker data. The long-term goal is to leverage these single studies through collaboration and coordination in order to address replication, extend findings to new contexts, and identify important factors that moderate healthspan and lifespan. The overall objectives of this application are to address critical human and technological barriers to collaboration and coordination in these deeply phenotyped studies. The central hypothesis is that these barriers relate to meta-data sharing (inputs) and multi-study analysis (outputs). The rationale for this project is that addressing “input” barriers such as PI reluctance and study team burden of meta-data sharing and “output” barriers such as difficulty accessing data and lack of training in multi-study analysis will lower the burden for successful coordination across studies, address broader replication questions, and empower smaller studies to fuel discoveries beyond their initially funded aims. These objectives will be pursued by four specific aims: development of both technological infrastructure (i.e., central distribution hub with publicly available meta-data catalog and collaboration/coordination resources [Aim 1]) and human infrastructure (i.e., incentivizing study PIs and engaging early career researchers [Aim 2], methodological support through workshops [Aim 3.1] and consulting [Aim 3.2], and financial support [Aim 4] for multi-study analysis). The proposed research is innovative, in the applicants’ opinion, because it develops tools and provides resources that reduce barriers for both PIs (e.g., dashboard for tracking and attribution, incentivizing meta-data sharing) and early career analysts (e.g., cross-study search and comparison tools, streamlined data requests). The proposed project is significant because the resulting collaborations and multi-study analyses will systematically test whether findings hold when tested in a diversity of sample characteristics, conditions, and across time. Ultimately, this will provide more rigorous tests of aging theories and their boundary conditions, which will improve understanding of aging and health.

NIH Research Projects · FY 2025 · 2025-05

Project Summary C. elegans is amenable to easy genetic modifications, as proven by its pioneering and continued use of endogenously expressed fluorescent reporters and other genetically encoded molecular tools. CRISPR/Cas systems for generation of mutants and transgenic lines are widely used in C. elegans. While highly precise, CRISPR/Cas applications are severely limited by the need to microinject nucleic acids to the germline, a significantly time-consuming process that drastically hinders the use of C. elegans for high-throughput biology. On the other hand, C. elegans has the ability to uptake double stranded RNA by feeding, which has led to the generation of dsRNA-expressing bacterial libraries and their use for large-scale reverse genetic screens. In this project, we aim to address this important limitation in C. elegans by developing a strategy that enables microinjection-free CRISPR/Cas applications. We will develop an approach to efficiently deliver RNA by feeding throughout C. elegans tissues for use as single-guide RNA (sgRNA) in CRISPR/Cas systems. Our approach relies in using easily uptaken RNA carriers to facilitate efficient import of sgRNAs through ingestion. We propose to address this challenge by: 1) studying uptake of diverse RNA structures into different C. elegans tissues through the development of an RNA sensor strain, 2) designing RNA structures for optimal uptake using artificial intelligence, and 3) testing the efficacy of modified sgRNA structures in CRISPR/Cas applications. Success of this work will greatly facilitate implementation of CRISPR/Cas approaches and make large-scale high-throughput CRISPR screens possible.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY: In this project we propose to develop and validate new ultrasound hardware technologies for high-contrast, high- resolution microvascular imaging to detect abnormalities in vascular morphology which can be biomarkers for a variety of disease ranging from cancer to dementia. Computed tomography angiography (CTA), photoacoustic microvascular imaging (PMI), and magnetic resonance angiography (MRA) have been used to image the micro- vasculature. However, it is highly desired to image the microvasculature using ultrasound alone. Ultrasonic Dop- pler imaging, acoustic angiography (AA), and ultrasound localization microscopy (ULM), which utilizes contrast enhanced ultrasound (CEUS) and ultrafast ultrasound imaging, have been demonstrated to be capable of provid- ing high contrast to tissue as well as high spatial resolution when imaging the microvasculature. Implementation of AA has long been challenged by the availability of transducer arrays that have a sufficiently wide bandwidth to excite microbubbles at low frequencies (LF) and receive the bubble generated harmonics at high frequencies (HF). In this project, we propose a novel capacitive micromachined ultrasonic transducer (CMUT) array structure, which is composed of a stack of a LF transmit (TX) elements and HF receive (RX) elements, fabricated one on top of the other, by using lithography-based microfabrication techniques. This approach allows independent op- timization of LF transmitters and HF receivers in the same physical area, hence resulting in entirely overlapping TX and RX apertures. The ability to independently design the TX and RX elements enables a large separation between the TX and RX frequency bands with no spectral overlap. Our long-term goal is the clinical translation of AA to enable imaging of morphology and function of microvasculature for improved diagnostics and therapy monitoring for pathologies including stroke, cancer, and dementia. The specific objective of this developmental project is to design and implement a custom imaging probe that includes the described duplex, dual frequency 1D CMUT arrays and supporting custom frontend electronics, all validated in small animal models. As the pro- posed project requires a high degree of innovation in array design and fabrication with potential results that can transform acoustic angiography to clinical space, the R21 mechanism is ideal to establish basic feasibility. Spe- cific aims of the project are as follows: 1) Design and implement duplex dual frequency 1D CMUT arrays with 32 TX elements operating in the 1-3 MHz band, and 256 RX elements operating in the 10-30 MHz band. 2) Design and implement custom frontend electronics combining linear TX drivers and low-noise wideband RX preamplifi- ers. 3) Validate and characterize the device in-vivo imaging performance in small animal models. The described CMUT structure with two arrays, i.e., LF and HF, microfabricated as a stack has never been proposed. Further- more, we innovate in the frontend electronics design to generate LF transmit pulses with minimal harmonic content to maximize the contrast-to-tissue ratio in the images. Our team has been collaborating on this significant application for over 5 years and therefore is in a great position to successfully execute the proposed project.

NIH Research Projects · FY 2026 · 2025-02

R-loops (RLs) are triple-stranded nucleic acid structures consisting of an RNA-DNA hybrid and a displaced single-stranded DNA loop. While unregulated RL accumulation causes genomic stress, DNA damage-induced RLs can also regulate the DNA damage response (DDR). While excessive telomeric R-loop (TRL) accumulation is linked to telomere shortening, and thus, aging, mechanistic understanding of the function of TRLs in the DDR at telomeres is limited. Recent findings from our lab and my co-sponsor, Dr. Li Lan (Duke University), implicate the cohesin-NIPBL complex and its cofactor, CCCTC-binding factor (CTCF), in TRL- mediated repair at telomeres. Our preliminary atomic force microscopy (AFM) data establishes cohesin and CTCF as avid R-loop binding proteins, and we show cohesin’s SA2 subunit binds to TRLs. Using a system that induces site-specific R-loops in a damage-dependent fashion in human cells (KillerRed system), Dr. Lan’s lab shows that cohesin recruits the RAD51 repair protein to genomic damage sites through R-loops. Additionally, they show cohesin’s SA1 and SA2 subunits localize to TRLs induced specifically at telomeres. Our preliminary data raises the possibility that cohesin and CTCF facilitate the telomere DDR by recognizing DNA damage- induced TRLs and recruiting repair proteins to damaged telomere sites. In this proposed work, I will test this hypothesis by leveraging a unique combination of in vitro AFM imaging and fluorescence cellular imaging with the KillerRed system. In Aim 1, taking advantage of the nanometer resolution of AFM, I will characterize the molecular mechanism by which cohesin and CTCF interact with TRLs by identifying the cohesin subunits involved in TRL binding and the role of CTCF in TRL localization by cohesin. In Aim 2, I will use a telomere- specific KillerRed system that induces localized DNA damage and TRLs at telomeres to determine the function of cohesin and CTCF in TRL-mediated DNA repair at telomeres in human cells. I will determine if damage- induced TRLs enhance cohesin and CTCF localization to damaged telomeres. Furthermore, I will determine if cohesin and CTCF recruit repair proteins to damaged telomeres via TRLs using multicolor fluorescence imaging. I predict that DNA damage-induced TRLs will be resolved upon completion of DNA repair mediated by cohesin and CTCF, and compromised recruitment of cohesin and CTCF at TRL will lead to DNA repair defects and TRL accumulation. To test this hypothesis, I will assess if the reduction in cohesin and CTCF levels with siRNA promotes TRL accumulation. Lastly, persistent telomere damage is known to accelerate telomere shortening and, with it, cellular aging. I will assess if cohesin and CTCF maintain telomere length in a TRL- dependent fashion by monitoring telomere lengths in human cells with knockdown of cohesin or CTCF. Through the proposed work, I will decipher a previously unexplored function of cohesin and CTCF in TRL- mediated DNA repair at telomeres, opening a new window into the world of telomere instability and aging.

NIH Research Projects · FY 2026 · 2024-12

ABSTRACT IgG Antibody (Ab) based therapeutics are becoming more common in treating several diseases, including infections, autoimmune disorders, transplant rejection, and cancer. Abs contain two important domains; the Fab domain mediates binding to a specific antigen, and the Fc domain of an antigen-bound IgG Ab then binds to Fc-gamma receptors (FcRs) expressed on the surface of different immune cells (e.g. NK cells, monocytes) and thereby initiates a broad array of effector functions related to target cell destruction, such as antibody- dependent cellular cytotoxicity (ADCC) and antibody-dependent cell-mediated phagocytosis (ADCP). However, Ab-based therapies are complicated by what appears to be suboptimal efficacy in a significant number of treated patients. A potential cause for this variability in response to Ab treatment could lie in genetic differences in the FcR locus, which is highly diverse genetically, with (at least) > 20,000 reported single nucleotide polymorphisms (SNPs) plus additional copy number variations (CNVs). Considering the extreme genetic complexity of the FcR locus, we have established a high-throughput assay to systematically screen for functional SNPs in the FcR locus. Based on the existing literature and our own results, we hypothesize that FcR polymorphisms modulate Ab functions across individuals, i.e., the same Ab but with different efficacy, by differentially regulating expression patterns of FcRs in a cell type-specific manner. Here we aim to systematically identify the functional impact of genetic variations in the FcR locus. Aim 1: Identify functional SNPs in the FcR locus that regulate expression of FcRs in human effector cells, and regulatory proteins binding to each SNP. Aim 2: Use the identified FcR SNPs and interacting proteins to derive predictive models for ADCC and ADCP activities. Ab based immune suppressions are routinely used to prevent transplant rejection and often function through ADCC and ADCP based mechanisms. We will enroll a group of 100 transplant patients in parallel with Aim 1, collect blood samples before transplant and measure their ADCC and ADCP activities in vitro. We will build computational models that predict ADCC and ADCP activities in individuals using their FcR genotypes alone, and together with other variables. Aim 3: Determine the impact of FcR genotypes on the efficacy of Ab based immune suppression regimens. After receiving Ab based depletional induction for transplantation, we will assess the contributions of these patients’ FcR genotypes and additional variables to the differences in their depletion efficiency. The expected outcome is to systematically identify functional SNPs in the human FcR locus and their impact on Ab based immunosuppression. This knowledge will further contribute to differentiating mechanisms of antibody-mediated rejection in transplantation and its treatment.

NIH Research Projects · FY 2026 · 2024-12

The overarching goal of the GenX Cohort Study is to answer community questions about the potential health effects of GenX and other per- and polyfluoroalkyl substances (PFAS). The GenX Cohort Study represents a unique opportunity to understand the impacts of PFAS throughout the Cape Fear River Basin (~ 9,000 square miles and 1 million residents). The central goal for this proposal is to provide a framework to strengthen and maintain the GenX Cohort Study infrastructure so that we can continue to provide timely information to impacted communities. In 2020, we were funded to create a community-based cohort of residents from the lower Cape Fear River Region (New Hanover and Brunswick Counties), Fayetteville, and Pittsboro as part of the NC State Superfund Center. A total of 1,091 individuals ages 6 and older were recruited; they provided blood samples for analysis of PFAS and clinical outcomes (lipids, thyroid hormones, and metabolic panel). With this grant, we will enhance our community engagement to ensure long term participation in the study. In efforts to maintain interest in the study, we will expand our community engagement throughout the region with the assistance of community organizations and local NGOs focusing on water and environmental protection. We will also build the infrastructure for the cohort by strengthening the coordinating center and data and specimen management activities. We will continue cohort follow up for another five years.

NIH Research Projects · FY 2025 · 2024-12

Project Summary/Abstract The overarching goal of the GenX Cohort Study is to answer community questions about the potential health effects of GenX and other per- and polyfluoroalkyl substances (PFAS). The GenX Cohort Study represents a unique opportunity to understand the impacts of PFAS throughout the Cape Fear River Basin (~ 9,000 square miles and 1 million residents). The central goal for this proposal is to provide a framework to strengthen and maintain the GenX Cohort Study infrastructure so that we can continue to provide timely information to impacted communities. In 2020, we were funded to create a community-based cohort of residents from the lower Cape Fear River Region (New Hanover and Brunswick Counties), Fayetteville, and Pittsboro as part of the NC State Superfund Center. A total of 1,091 individuals ages 6 and older were recruited; they provided blood samples for analysis of PFAS and clinical outcomes (lipids, thyroid hormones, and metabolic panel). With this grant, we will enhance our community engagement to ensure long term participation in the study. We will prepare materials in Spanish and maintain a Spanish language website. In efforts to maintain interest in the study, we will expand our community engagement throughout the region with the assistance of community organizers and local NGOs focusing on water and environmental protection. We will also build the infrastructure for the cohort by strengthening the coordinating center and data and specimen management activities. We will continue cohort follow up for another five years and work to build a more inclusive workforce in environmental health and epidemiology.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY To address the public health issue of voice disorders affecting approximately 20 million Americans annually due to vocal fold (VF) scarring and laryngotracheal injuries, this project focuses on a non-invasive, innovative treatment strategy. Utilizing a specially derived Vocal Fold Lamina Propria extract (VFLPx), which has demonstrated promising results in reducing fibrotic gene expression, the proposed project aims to fine-tune the dosage and application methods of VFLPx. The research is structured around three principal objectives: (1) To conduct a detailed investigation into the anti-fibrotic and anti-microbial properties of VFLPx through advanced proteomic analysis and targeted assays, identifying the key molecular constituents responsible for its therapeutic effects. (2) To assess the efficacy of VFLPx in promoting wound healing within a rabbit model of VF injury, exploring its use both as a singular treatment option and in conjunction with aerosolized forms, aiming to minimize the reliance on invasive surgical interventions. This will include evaluating the benefits of both direct injection and aerosolized delivery to determine the most effective approach for tissue repair. (3) To rigorously evaluate the safety of nebulized VFLPx in a rat model, particularly its impact on lung function and the respiratory system, to ensure its suitability for long-term use in human patients. The project’s ultimate goal is to establish a solid foundation for developing a less invasive, more patient-friendly approach to treating VF and laryngeal injuries, potentially revolutionizing the management of such conditions and significantly improving patient outcomes.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY Spatially resolved ‘omics techniques have been instrumental in describing the heterogeneity in molecular content in tissues. However, these methods can only provide static measures of the variation in content and fall short on relating how this molecular heterogeneity results in differences in tissue function. Our laboratories have developed a unique functional Mass Spectrometry Imaging (fMSI) method that can measure tissue perfusion and metabolic activities at high spatial resolution and can provide a critical link between spatially resolved ‘omics data and cellular phenotype. This fMSI method uses timed infusions of isotopologues of a single substrate prior to tissue harvest to obtain dynamic data at each sampled location. We demonstrated the feasibility of this method to detect uptake (perfusion) of isotopologues of glycine and their conversion to glutathione in liver and tumor tissue but there remain significant challenges to realizing the full potential of this method. This project is centered around four aims. Aim 1 will evaluate different bolus and constant infusions protocols for isotopologue administration with the objective to improve the sensitivity and extend the observable metabolic window. Aim 2 will administer isotopologues of pimonidazole and urea to demonstrate that fMSI can detect oxygenation and perfusion dynamics in tissues. Aim 3 will demonstrate the general utility of the fMSI method to measure pentose phosphate pathway activity in tissues. Aim 4 will tie together the fMSI data to generate spatially-specific mathematical models of tissue metabolism and validate in vivo observations of the effect of microenvironmental variations on metabolism using in vitro perfusion models. Successful completion of these Aims will position fMSI as a unique bioanalytical tool to provide high spatially resolved functional data in any tissue sample.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Disseminated intravascular coagulation (DIC) is the pathophysiologic response to injury that leads to the formation of microthrombi and subsequent activation of fibrinolysis. DIC is characterized by the consumption of platelets, fibrinogen, and clotting factors, and it can present clinically with prothrombotic and/or hemorrhagic phenotypes. DIC is most commonly caused by sepsis but can also be caused by trauma, cancer, pregnancy, or severe organ injury. The treatment approach for DIC involves addressing the underlying cause, but the contradictory nature of the disorder makes identifying an effective treatment protocol challenging, which is reflected in the high mortality rate (50%) for humans with sepsis-induced DIC. DIC does not only affect humans however, and the mortality for dogs with DIC is even higher at 62.5%. The benefits of anticoagulants and fibrinolytics are still being studied but have been associated with an increased risk of off-target bleeding. We have developed fibrin-specific nanogels (FSNs), which we have used in vivo to site-specifically deliver anti- clotting drugs while also enhancing fibrin cross-linking and clot formation. Preliminary data have shown that anticoagulant- or fibrinolytic-loaded FSNs address both DIC phenotypes by targeting and dissolving existing clots, while simultaneously promoting clotting at sites of bleeding. Notably, the drug-loaded FSNs improve hemostasis within only thirty minutes. When dual-loaded with both fibrinolytic tissue plasminogen activator (tPA) and anticoagulant antithrombin-3 (AT3), FSNs have the potential to provide an enhanced effect by acting on two different mechanisms. The objective of this proposal is to establish anticoagulant- and fibrinolytic-loaded FSNs as safe and effective adjuvant therapies for sepsis-induced DIC and to further understand the mechanisms by which loaded FSNs rapidly improve DIC outcomes. It is expected that dual-loaded FSNs will result in better DIC outcomes than tPA and AT3 administered systemically or individually in FSNs by interacting directly with neutrophils to recover platelet count and reduce hemorrhage. Aim 1 will optimize FSN drug-loading by examining different loading doses of tPA and AT3 in FSNs and assessing their stability, loading efficiencies, release profiles, and clotting and fibrinolytic abilities. Aim 2 will optimize the dose of loaded FSNs in vivo by examining biodistribution, coagulation, microthrombi development, and side effects. Aim 3 will assess the role of neutrophils in rapidly rescuing platelet counts with in vitro and in vivo models treated with drug-loaded FSNs. The successful completion of these aims will identify the pathophysiology behind targeted tPA and AT3 treatment and provide the framework for moving this novel DIC therapy into clinical trials. This proposal and associated mentorship team will provide training critical for the applicant’s development as an independent clinician-scientist developing drug delivery platforms for cardiovascular diseases that affect both humans and animals. The training environment at North Carolina State University is ideal for the applicant’s interdisciplinary training goals, as the applicant also has access to resources and opportunities at Duke University and UNC-Chapel Hill.

NIH Research Projects · FY 2024 · 2024-09

PROJECT ABSTRACT Osteoarthritis (OA) is a major and growing public health problem that negatively impacts quality of life. The prevalence of painful arthritis in the U.S. likely approaches ~90 million adults and clinical sequela of OA-associated pain include decreased mobility and compromised activity. Importantly, the chronic pain experience in humans frequently includes profound and debilitating effects on the emotional state, with significant negative impacts on quality of life and function. Consequences also include impaired performance on cognitive tasks, particularly those requiring working memory or attentional switching. These effects compound the clinical picture, contributing to the pain-related depression, anxiety, and emotional distress (the `experience' of chronic pain). Research relying on rodent models is not translating into new, effective treatments. One reason for the lack of translational success is that despite the clear importance of emotion and cognition in the human chronic pain experience, current models of chronic pain in animals frequently ignore these critical domains. The major goal of the proposed studies is to bridge this `model gap' and significantly advance translational research capability by developing and rigorously validating a battery of assays for assessment of emotions and cognitive function in the pet dog model of persistent OA pain. Pet dogs with naturally occurring persistent OA pain are already considered a good model of the sensory- discriminative aspects of OA pain in humans; enhancing the capability of this model will allow researchers, for the first time, to access a clinically relevant full biopsychosocial animal model of persistent pain. We will achieve this through developing, refining and rigorously validating (test-retest, structural, discriminative, responsiveness, and criterion validity) a battery of emotional and cognitive domain tests, benchmarking against validated measures of pain and the impact of pain. Applying advanced statistical techniques, we will create a concise battery that can be feasibly performed in clinical research settings. We bring together diverse expertise with proven track records of collaboration and established facility resources to successfully address this critical gap in modeling the pain experience of humans. Successful completion of this proposed work will validate a highly clinically relevant biopsychosocial animal model of persistent musculoskeletal pain that has the potential to radically increase the translation of pre-clinical knowledge into effective, non-addictive analgesic treatments for humans suffering from persistent musculoskeletal pain.