Virginia Polytechnic Inst And St Univ

NIH Research Projects · FY 2026 · 2026-06

SUMMARY – Lina Ni The long-term goal of my lab is to identify how animals detect various environmental stimuli to regulate their behavior. Temperature and moisture are closely interrelated environmental variables that animals continuously encounter; animals must regulate and maintain their temperature and moisture homeostasis to sustain normal physiological functions. Optimal environmental conditions bolster development and reproduction, whereas suboptimal conditions negatively impact these processes, and extreme conditions may lead to rapid mortality and other life-threatening conditions. Temperature and moisture variations are particularly significant for insect population, distribution, and behavior. Both larvae and adults rely on ambient temperatures to set their body temperature and on environmental moisture conditions to maintain their hydration state. These two environmental variables also determine the developmental rate and success of larvae, and unfavorable conditions challenge insects’ immune systems and increase their susceptibility to pathogens. Therefore, temperature and moisture are directly linked to the outbreak of insect disease vectors and agricultural pests. This proposal uses fly larvae as a model system to study how insects detect and integrate temperature and moisture information. Fly larvae possess molecular receptors for these senses that are evolutionarily conserved to those found in other insects. They can detect subtle changes in temperature and moisture and guide robust behaviors toward optimal conditions. Their transparent bodies, short lifespans and generation times, simple nervous systems, and powerful genetic tools enable precise manipulation and visualization of molecular receptors, sensory neurons, and neural circuits. Despite these advantages, the mechanisms of moisture sensing in larvae remain unknown. It is also unclear how animals integrate temperature and moisture information at the molecular, cellular, and circuit levels to guide navigation toward optimal environments. The goal of this proposal is to determine how fly larvae sense and integrate temperature and moisture information in sensory neurons and in the brain. Successful completion of this proposal will identify the larval moisture-sensing mechanisms and uncover how temperature and moisture sensation modulate each other. This work will enhance our understanding of insect sensory biology, evolution, and behavior and support novel strategies to predict outbreaks and control insect disease vectors and agricultural pests.

NIH Research Projects · FY 2026 · 2026-06

Abstract: The management of invasive fungal infections remains a significant challenge due to the limited arsenal of antifungal drugs and the growing resistance among fungal pathogens. Currently, only three main classes of antifungal agents (azoles, polyenes, and echinocandins) are used to treat these infections. Resistance to azoles has become alarmingly prevalent among fungal pathogens. While AmB is effective against most fungal pathogens, its clinical application is hampered by severe side effects and dose-limiting toxicity. Alarmingly, approximately 30% of the emerging fungal isolates, Candida auris, exhibit resistance to AmB and certain fungal pathogens are intrinsically resistant to echinocandins, further narrowing therapeutic options. The discovery of novel antifungal drugs remains very difficult, emphasizing the need for innovative strategies to combat drug resistance. This proposal outlines a milestone-driven, early-stage translational research project focused on the discovery and development of novel therapeutics targeting select fungal pathogens, including Candida auris, Aspergillus fumigatus, and Mucorales. Specifically, the research aims to explore the potential of Neurokinin-1 (NK1) receptor antagonists as antifungal agents. This work aims to establish NK1 inhibitors, synthesize chemical analogs, optimize their safety and pharmacokinetic properties as novel antifungal therapies, offering a promising solution for the treatment of life-threatening fungal infections while addressing the pressing issue of antifungal resistance.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract The broad goal of this proposal is to understand how obesity interacts with the aging process and accelerates memory decline across the lifespan. Age-related memory loss occurs in ~33% of adults over the age of 70, yet no treatment option exists that can prevent or reverse these impairments. One of the major risk factors for developing age-related memory impairments is obesity, which affects nearly 40% of U.S. adults. However, even though obesity and aging are both known to be associated with cognitive decline, very little is known about how they interact to affect memory across the lifespan. The ubiquitin-proteasome system (UPS) controls the majority of protein degradation in cells and is dysregulated with age and in Alzheimer’s disease. However, the ubiquitination process is complex, and few studies have examined the role of degradation-independent ubiquitin modifications in the brain. K63 polyubiquitination is the second most abundant form of ubiquitination and is independent of protein degradation. We recently found that reductions in K63 polyubiquitination the hippocampus is critical for normal memory formation in young adulthood. Interestingly, we also found that both aging and obesity in young adult rats leads to aberrant increases in K63 polyubiquitination in the hippocampus, the primary brain region involved in long-term memory formation, which correlates with obesity- and age-related memory impairments. Further, knockdown of K63 polyubiquitination in the aged hippocampus rescues age-related memory deficits. However, it is unknown whether the obesity-induced and age-induced K63 polyubiquitination dysregulation are directly connected or instead just share some common molecular mechanisms. The work in this proposal is designed to test 1) if obesity accelerates age-related memory decline via dysregulation K63 polyubiquitination in the hippocampus, and 2) whether reducing aberrant increases in K63 polyubiquitination in the hippocampus can prevent obesity- and aging-induced memory decline. Using a diet-controlled longitudinal design in combination with sophisticated protein purification methods and unbiased mass spectrometry whole proteome analyses, Aim 1 will test if obesity accelerates the aging process in the hippocampus or instead results in a unique process characterized by dysregulated K63 polyubiquitination of a distinct subset of proteins. Aim 2 will use a diet-controlled longitudinal design in combination with cutting-edge CRISPR-dCas13-mediated knockdown of hippocampal K63 polyubiquitination to test if preventing obesity-induced and aging-induced increases in K63 polyubiquitination can preserve memory across the lifespan. Collectively, this study will answer important questions about how obesity can accelerate age-related memory decline and increase the susceptibility to age-associated neurodegenerative disorders. Results from this project could provide critical insights that may lead to the development of novel treatment strategies designed to mediate the effects of obesity on the aging process

NIH Research Projects · FY 2026 · 2026-05

The protozoan parasite Toxoplasma gondii infects approximately one-third of the world’s population. Most people are asymptomatic because the parasite resides latently within the brain and other tissues. But AIDS and other immunosuppressed patients as well as fetuses are in danger of developing toxoplasmosis when the parasite reactivates and the host immune response is unable to control parasite replication or is dysregulated. When reactivation occurs in the brain, toxoplasmic encephalitis can develop and present with a variety of neurological complications that includes seizures. Why patients develop seizures is unclear but in other encephalopathies seizures develop due to a variety of reasons including breakdown of the blood-brain barrier, changes in ionic homeostasis, and/or increased abundance of inflammatory proteins such as cytokines and antibodies. And, how these changes in the brain alter synaptic wiring and neurotransmission that cause epileptiform activity is largely unknown. Data from a murine toxoplasmic encephalitis model reveal that onset of seizures is associated with three events – increased interleukin-1 signaling, increased excitatory neurotransmission, and ensheathment and subsequent loss of inhibitory perisomatic synapses. The work proposed here will seek to understand how loss of inhibitory perisomatic synapses occurs. In Aim 1, we will study how the complement system recognizes the inhibitory synapses to be eliminated and in Aim 2 we will identify the factor(s) that neurons express to recruit microglia that remove these synapsesAddressing and answering these questions will increase our understanding of how pathogen-host cell interactions lead to seizures and will inform future work aimed at generating novel therapies to treat these patients .

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Heart failure is a leading cause of death in developed countries, affecting 6 million people in the United States and approximately 64.3 million people worldwide. Mortality rates remain high, with 50% of patients dying within five years of diagnosis. About half of these patients have heart failure with reduced left ventricular ejection fraction (HFrEF), which is characterized by an ejection fraction of 40% or less and adverse cardiac remodeling. HFrEF is associated with reduced cardiac contractility and mitochondrial impairment. Currently, no therapies specifically target both muscle contractility and mitochondrial function. PERM1, a striated muscle-specific regulator of energy metabolism, is predominantly expressed in the heart and skeletal muscle. We have demonstrated that PERM1 is downregulated in both human and mouse failing hearts and that its loss in mice leads to reduced cardiac contractility and energy reserves. Our preliminary data show that gene delivery of PERM1 to the mouse heart via adeno-associated virus (AAV) fully prevents the onset of HFrEF during pressure overload in mice, preserving systolic function and promoting mitochondrial biogenesis. This study will address the hypothesis that AAV-medicated PERM1 overexpression can serve as a novel therapeutic approach to prevent and reverse contractile dysfunction under pathological stress. This translational study will initiate testing of this promising gene therapy in a well-established mouse model of HFrEF and a preclinical swine model of HFrEF induced by myocardial infarction. Additionally, it will further elucidate the novel mechanisms by which PERM1 simultaneously regulates muscle contractility and energy metabolism in the heart.

NIH Research Projects · FY 2026 · 2026-04

Sepsis is a leading cause of death worldwide, with most patient mortality stemming from lingering immune dysfunction in sepsis survivors. A key feature of sepsis-associated immune dysregulation is monocyte exhaustion, a phenotype of paradoxical pro-inflammatory and immunosuppressive gene expression, impaired differentiation, and reduced antigen presentation. Monocyte exhaustion can persist for years after sepsis onset, a result of long-term immune memory. However, the mechanisms controlling such long-term memory remain to be elucidated. Whereas previous research has conceptualized innate immune memory through diametrically opposed mechanisms that either promote (train) or restrict (tolerize) monocyte responses, my preliminary data suggests that exhaustion represents a distinct memory state characterized by unique immune, transcriptional, and epigenetic features. Therefore, in contrast to the two-state model for innate memory, I hypothesize that innate memory represents a continuum of states driven by distinct epigenetic patterning, with prolonged, high- intensity immune stimulation leading to monocyte exhaustion in septic individuals. In Aim 1 of my proposed study, I will profile the unique transcriptional and epigenetic features defining monocyte exhaustion, as well as employ integrative modeling to determine how immune stressor strength, duration, and timing influence the establishment of distinct innate memory states. In Aim 2, given preliminary data showing genome-wide DNA hypermethylation in exhausted monocytes, I will test the hypothesis that inhibition of DNA demethylation enzyme TET2 is upstream of these epigenetic changes, and that treatment with TET agonists is a tractable therapeutic strategy to restore healthy epigenetic memory. Finally, in Aim 3, based on my recent identification of a novel DNMT3L isoform expressed in septic monocytes, I will test the altered chromatin affinity and regulatory activity of this isoform and establish its contribution to DNA methylation reprogramming during monocyte exhaustion. Completion of these proposed Aims will allow me to develop skills in new experimental techniques, including single-cell RNA sequencing, reduced representation bisulfite sequencing, in vivo mouse sepsis modeling, and cytometric arrays. Aims 1 and 3 will be pursued during the K99 mentored research phase at Virginia Tech in the laboratory of Dr. Liwu Li, an expert in the fields of monocyte biology and innate immune memory. Whereas my previous graduate studies focused on epigenetics and mammalian development, Dr. Li will provide valuable instruction as I expand into the topics of immunology and hematology. I will also pursue coursework at Virginia Tech in computational modeling of biological systems while engaging with professional development workshops covering such topics as scientific communication, mentorship, and R-series proposal development. The goal of this project is ultimately to pursue a career as an independent biomedical investigator in academic research; these studies will serve as a foundation for my own research program aimed at identifying the major molecular players responsible for establishing and maintaining innate immune memory.

NIH Research Projects · FY 2026 · 2026-04

Progress in modern bio-molecular sciences, from structural biology to structure-based drug design, is greatly accelerated by methods of atomic-level modeling and classical simulations that bridge the gap between theory and experiment; 45,000+ research papers that use these methods are published each year. Accurate and computationally facile water models are just as important for outcomes of these studies as water is for Life. In practice, several principal levels of compromise exist between level of detail and speed of solvent models. However, critical accuracy and performance gaps remain at each level, these gaps mute the strong potential of atomistic modeling. For example, even with most detailed (explicit) water models, significant discrepancies with experimental binding free energies are still seen, which is one critical factor that hampers rational drug design efforts. Another problem is computational cost, which can become prohibitive when most accurate existing models are used. On the other hand, in many areas, which can benefit from faster, less detailed (so-called “implicit solvent”) water models, simulations based on these faster models are often unreliable. New solvent models appear regularly, but these are often limited to re-parameterizations of old ones, or utilization of old “base models” to add key new features such as electronic polarization. My lab has always focused on ground up, physics-based approaches to model development, which are more likely than many alternatives to yield robust, transferable models that stand the test of time. The previous funding period has enabled us to accumulate a critical mass of innovations in the field of solvent model development, innovations that have already shown significant promise in practical applications. Importantly, the reported improvements in water model accuracy came without sacrificing the speed. The goal for the next 5 years is to move the entire field of atomistic simulations to the next level of predictive accuracy by delivering to the community a novel class of solvent models, at each key level of detail--speed compromise. To demonstrate utility of the new models (once thoroughly tested), we will apply them to: (1) Improving the accuracy, without sacrificing speed, of estimation of receptor-ligand binding free energies. In structure-based drug discovery, the accuracy and computational efficiency of in-silico binding free energy predictions for small molecules to biomolecular targets are crucial for high-throughput screening of drug candidates. (2) Generation of novel insights into regulation of DNA accessibility in the nucleosome, which directly affects gene expression. Understanding how DNA accessibility in the nucleosome is controlled/affected by various biologically relevant factors is a fundamentally important problem of direct biomedical relevance. The disruption of the histone functions leads to diseases. In addition, the novel, higher accuracy, yet efficient solvent models will be implemented into H++ web-server maintained by the PI (12,000+ registered users, 20,000+ requests per year), thus immediately improving outcomes of structure preparation and analysis efforts for a large modeling community.

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY This proposal is for a K01 Mentored Research Scientist Development Award for Mary Elizabeth Baugh, PhD, RD. Dr. Baugh’s long-term goal is to become an independent investigator studying the synergistic role of metabolic and neural signals on energy balance and eating behaviors and translating these insights into novel, individualized treatments for obesity and metabolic disease. Substantial evidence in both animal models and human neuroimaging studies highlights altered brain morphology, functional connectivity, and reward responses in obesity; however, it is much less clear whether these brain alterations map on to behavioral patterns of learning and decision-making. There is a growing appreciation for the impact of post-ingestive metabolic signals on brain regions involved in food reward and eating behaviors, and recent evidence suggests altered brain insulin activity disrupts regulatory mechanisms governing eating behaviors in reward and decision-making brain regions. Given that obesity and insulin resistance commonly co-occur, and that most central insulin is derived from peripherally circulating insulin, the central hypothesis of this proposal is that reward learning and decision-making are attenuated in obesity and that reduced insulin sensitivity influences this alteration. Data from Dr. Baugh’s pilot study support this hypothesis, suggesting that lower glycemic control is associated with reductions in behavioral reward learning. The aims of the proposed research are to assesses the influence of excess adiposity and insulin sensitivity on 1) post-ingestive reward learning using a flavor-nutrient learning paradigm, which is based on Pavlovian classical conditioning, and 2) decision-making for food and non-food rewards using probabilistic selection tasks paired with computational modeling of learning parameters. Dr. Baugh has an exceptional training background in clinical physiology and metabolism but requires protected time to gain expertise in theoretical concepts of appetitive and cognitive neuroscience and develop the necessary practical skills to launch her independent research program as a tenure-track faculty member. The mentorship team, training objectives, and research aims outlined in this proposal will provide an exceptional foundation for Dr. Baugh, and the Fralin Biomedical Research Institute at VTC is an ideal environment for such training. Her co-primary mentors, Drs. Alexandra DiFeliceantonio and Pearl Chiu have complementary expertise the neurobiology of eating behaviors and computational modeling of reward and decision-making, respectively. Dr. Baugh’s training plan will focus on 1) expanding her theoretical knowledge of neural and behavioral underpinnings of food reward and eating behaviors, 2) expanding her theoretical knowledge of computational neuroscience, 3) enhancing her skills in scientific rigor, reproducibility, and statistical analysis in clinical trials, and 4) enhancing her technical skills in functional magnetic resonance imaging (fMRI) study design, data acquisition, data procession, and data analysis. Overall, the trajectory of this work and Dr. Baugh’s career goals have the potential to guide more biologically informed, individualized strategies for treatment of obesity and metabolic disorders.

NIH Research Projects · FY 2026 · 2026-03

Project Summary Alphaproteobacteria are a diverse taxon and many are capable of motility using flagella. They comprise several genera, including model organisms such as the plant symbiont Sinorhizobium meliloti and the plant pathogen Agrobacterium tumefaciens. Flagellar motility is an invaluable colonization factor required by soil bacteria to navigate the rhizosphere and accelerate associations with their plant hosts. Research within my laboratory is centered on flagellar-driven motility. We discovered that rhizobia evolved complex regulatory networks that adjust their cellular physiology and control the assembly of a specially-adapted flagellar machinery to permit efficient navigation of the soil environment in search for nutrients and host plants. Specifically, S. meliloti employs a unidirectionally rotating, variable-speed flagellar motor that is compositionally distinguished from the well- characterized switch-type motors of enteric bacteria, enabling it to swim in highly viscous environments. Our studies also revealed that the diverse and rapidly varying soil environment and the metabolic diversity and specific adaptations of soil bacteria to host signals led to the evolution of complex chemosensory systems compared to those of enteric bacteria. Recently, we uncovered several unique components and features that control S. meliloti chemotaxis to direct the bacterium toward nutrient sources and hosts. Our significant progress in deciphering these complex pathways establishes S. meliloti as model for studying the architecture and evolution of bacterial chemosensory and motility systems. Bacteria use variations of the chemotaxis signaling cascade to control cellular development, but knowledge of their specific roles remains scarce. The functions of alternate chemosensory pathways are understudied because their activating signals are mostly unknown and difficult to identify, but this knowledge is crucial for studying their regulatory functions. As such, the role of the Che2 chemosensory system in S. meliloti is unknown. My research goals for the next five years are to decipher the underlying molecular mechanisms of the unidirectional, variable-speed flagellar motor that enables bacteria to move efficiently in the soil. The receptor modification system of S. meliloti plays a pivotal role in its chemotaxis- directed movement, yet, remarkably, little is known about the regulation of pathway sensitivity and stimuli adaptation, which are essential for an effective chemotactic response. We plan to investigate how S. meliloti retains its chemotactic memory and how it effectively senses attractants over a wide range of attractant concentrations. We also plan to identify environmental and/or host signals that activate the Che2 system and characterize its regulatory pathway, which will contribute to the knowledge of these alternate systems and their roles in cellular functions. The overall vision of my research program is to advance a fundamental understanding of rhizobial motility and cell physiology, which are important pre-requisites for survival under various challenges and, ultimately, host engagement. This research will reveal new processes that support optimal growth and inform about new concepts in signal transduction, which will be transformative to other bacterial systems.

NIH Research Projects · FY 2026 · 2026-02

Project Summary/Abstract. Arboviruses, such as Mayaro virus (MAYV), are emerging with severe impacts on human health. Initially identified in Trinidad and Tobago in 1954, MAYV is widespread in South America, causing outbreaks of febrile illness and chronic, debilitating joint pain. Due to similar symptoms with chikungunya, dengue, and other diseases, along with inadequate surveillance, MAYV infections are underreported despite seroprevalence rates exceeding 50% in some regions. MAYV is typically maintained in a sylvatic transmission cycle involving Haemogogus mosquitoes and various animals, but human-biting mosquitoes like Aedes aegypti, Ae. albopictus, Anopheles albimanus, and An. quadrimaculatus are competent vectors in the laboratory. The recent detection of MAYV in Haiti, an area lacking forests, suggests a shift to urban transmission involving human-biting mosquitoes. However, most studies on MAYV have used historical virus strains, not accounting for potential adaptations in contemporary strains. Our understanding of MAYV’s risk of expansion is therefore limited. We propose a proactive approach to evaluate the risk of contemporary MAYV strains. Recent findings of three distinct MAYV lineages in Haiti indicate possible evolutionary adaptations. Given the travel frequency between Hispaniola (the island encompassing Haiti and The Dominican Republic) and North America, the risk of further spread is significant. Here, we propose two aims: In Aim 1, we will develop novel molecular tools for studying MAYV and assess the transmission potential of contemporary, Haitian MAYV strains by widespread mosquito vectors, including field-caught mosquitoes. In Aim 2, we will evaluate the replication and virulence of contemporary MAYV strains in human cells and mice and assess MAYV transmission in a complete transmission cycle between mice and mosquitoes, determining the minimum infectious dose required to infect mosquitoes. This work is vital for understanding MAYV’s expansion potential. This study will identify differences between strains that can be used in future studies to identify viral genetic determinants driving changes in transmission or virulence. Finally, the insights gained will contribute significantly to global health, offering novel strategies and tools for arbovirus research.

NIH Research Projects · FY 2026 · 2026-02

Abstract: Clostridioides difficile is the leading cause of nosocomial infections and antibiotic-associated diarrhea. The incidence and severity of C. difficile infections (CDI) have dramatically increased due to the overuse of antibiotics and the emergence of hypervirulent epidemic strains, which were responsible for several outbreaks globally. Even though the overuse of antibiotics is responsible for CDI, the management of CDI requires antibiotic administration. Consequently, the drawbacks associated with the current anti-CDI therapeutic arsenal highlights the critical need for developing novel strategies for the treatment of CDI and the prevention of CDI recurrence. An alternative strategy for developing novel therapeutics involves the exploitation of antisense oligomers (ASOs), such as peptide nucleic acids (PNAs) that suppress essential genes in bacterial pathogens. This research project focuses on developing targeted therapeutics to disarm virulence factors and suppress gene expression in C. difficile. Building on our successful use of antisense technology in C. difficile, we aim to investigate the potential and implication of inhibiting critical pathways within C. difficile. These pathways include transcription and translation machinery, protein translocation machinery, cell division, replication, and fatty acid synthesis. We are also planning on assessing the toxicity and in vivo efficacy in murine models of CDI and its recurrence. We believe the narrow spectrum and precision targeting of these antisense therapeutics provide an innovative new approach that selectively eliminate problematic pathogens, such as C. difficile without harming the healthy human gut microbiota. Altogether, these studies are of paramount importance and have the potential to make a significant impact, effectively leapfrogging the traditional drug development process.

NIH Research Projects · FY 2026 · 2026-02

PROJECT SUMMARY/ABSTRACT The rising prevalence of Alzheimer's Disease (AD) poses a significant challenge for human health. Characterized by progressive memory loss and cognitive decline, the long-term care costs for AD patients in the U.S. is expected to reach a trillion dollars by 2050. Despite decades of clinical research, there are few FDA-approved drugs to treat or prevent AD. Thus, a greater understanding of the mechanisms contributing to AD memory loss are needed to identify promising therapeutic targets. The entorhinal cortex (EC) is an initial site of tau and amyloid pathology in Alzheimer's patients and AD model mice. The major EC output is to the hippocampus, and EC-hippocampus synapse loss is a presumed cause of early memory impairment in AD. However, the molecular basis of EC circuit vulnerability is unknown. Along with synapse loss, mitochondrial dysfunction is an early pathological feature of AD. Because mitochondria provide fuel and metabolites vital for synapse function, restoring mitochondria and thereby synapse function may prevent or delay AD progression. Hippocampal CA2 mitochondria in ECII-contacting distal dendrites are larger than mitochondria in ECIII- contacting CA1 distal dendrites. Further, the mitochondrial calcium uniporter (MCU) is enriched in dendritic mitochondria near ECII-CA2, but not ECIII-CA1, synapses. Because excessive mitochondrial calcium is a proposed driver of AD pathology, we hypothesize that ECII-CA2 mitochondria and synapses are more vulnerable to AD pathology due to excess dendritic MCU-mediated calcium uptake. The ECII-CA2 synapse has yet to be investigated in the context of AD, and its distinct mitochondrial population is an ideal model to study the crosstalk between dendritic mitochondria and synapse function in the context of dysregulated mitochondrial calcium signaling in AD. In this Next Generation R03 proposal, we use acute slices from control and amyloid-based AD model mice to directly compare mitochondrial calcium uptake and mitochondrial fragmentation in EC-contacting CA2 and CA1 dendrites (Aim 1) as well as ECII-CA2 and ECIII-CA1 synapse function when mitochondrial calcium uptake is blocked (Aim 2). Results from these aims will reveal whether MCU-enriched dendritic mitochondria confer EC circuit vulnerability in an amyloid-based AD model. A new AD research program based on the outcome of this pilot study will identify the upstream mechanisms underlying dysregulated mitochondrial calcium signaling as well as the downstream consequences on mitochondria products and synaptic receptors essential for synapse function in amyloid- and tau-based AD models.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Alcohol use disorder (AUD) is a chronic disease that affects approximately 30 million people, resulting in $250 billion in lost productivity, and associated healthcare costs. Although three FDA-approved medications are available to treat AUD, achieving long-term abstinence remains challenging, indicating new therapeutic approaches would be beneficial. The cocaine and amphetamine regulated transcript is a neuropeptide involved in several physiological processes, including AUD. However, studies of CART II during AUD have been prevented by the lack of a known receptor. We recently identified the Lysophosphatidic Acid Receptor 2 (LPAR2) as a high affinity receptor for CART II in the brain. We used cell-based assays and in vivo pharmacology tools to show that LPAR2 is mediating CART II behavioral effects. Our preliminary data show that brain administration of CART II during intermittent alcohol 2-bottle choice (IA2BC) reduces ethanol intake in wild-type male (WT) without altering water or sucrose consumption. However, the effects of CART II on escalated drinking, a hallmark of AUD, have not been investigated. Previous work showed that during alcohol withdrawal extracellular glutamate (GLU) levels in the nucleus accumbens (NAcc) are elevated and this contributes to neuroadaptations occurring in the medium spiny neurons (MSNs). Our preliminary data demonstrates that LPAR2 is expressed in the NAcc and confirms LPAR2 couples with Gi/o protein suggesting CART II signaling may decrease GLU vesicular release in the NAcc and/or decrease the excitability of D1-MSNs. However, the presynaptic and postsynaptic effects of CART II-LPAR2 signaling in the NAcc have not been established. The overarching hypothesis of this proposal is that CART II – via LPAR2 – reduces escalated ethanol drinking and reverses the ethanol-induced neuroadaptations. Thus, we propose to use the well-established chronic-intermittent ethanol vapor exposure interspersed with two-bottle choice (CIE-2BC) model in WT and LPAR2 KO mice of both sexes to fully examine the effects of CART II-LPAR2 signaling on escalated alcohol intake (Aim 1), the pre-synaptic and post-synaptic effects on D1-MSNs excitability in the NAcc (Aim 2) and CART II region-specific effects on GLU release in the NAcc (Aim 3). To validate that the effects of CART II on AUD are mediated by LPAR2, we will employ both sexes and LPAR2 KO mice. Collectively, the expected results will elucidate the CART II-LPAR2- mediated mechanism on AUD, facilitating the development of novel therapeutics. An experienced team of mentors and career advisors will provide training critical for the candidate’s short- and long-term success. The PI will be trained at Virginia Tech (VT) on the design, execution, and interpretation of behavioral models of alcohol use, and ex vivo electrophysiology of distinct subpopulations of neurons (both at VT and Scripps Research Institute). Finally, the professional development plan will prepare the candidate to transition to an independent position and create her future research program in studying the interaction between neuropeptides and NTs in multiple neuronal subpopulations during AUD and other substance use disorders.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY There is a critical gap in understanding the health impacts of inland flooding, particularly in rural Health Professional Shortage Areas (HPSAs). These challenges are currently hindering ongoing recovery efforts following Hurricane Helene in rural Appalachian areas of North Carolina, Tennessee, and Virginia. While previous studies demonstrate increased pregnancy complications and mental health issues following flooding events, the disease burden, barriers to healthcare utilization, and exposure pathways are less studied, particularly in rural areas already grappling with a shortage of mental health services and the consolidation or closure of obstetric units. These gaps prevent effective planning and resource allocation and exacerbate uncertainties around the effectiveness of preparedness strategies designed to minimize the effects of future flooding. Collecting time-sensitive data will support recovery efforts and guide future planning and preparedness strategies. Our team has developed methods to rapidly collect data from healthcare delivery organizations, local agencies, recovery organizations, and school staff to document challenges in meeting health-related needs and identify priority populations and outcomes. We propose to undertake three aims. In the R61 phase, we will identify critical health-related barriers encountered during the Helene response and recovery efforts in affected areas of North Carolina, Virginia, and Tennessee, and we will support ongoing recovery via knowledge exchange sessions with health systems in Kentucky and West Virginia that experienced major floods in 2022 and 2016, respectively. In the R33 phase, we will integrate R61 collected data with healthcare visit data and implement a quasi-experimental controlled before and after study design to estimate flood-related changes in overall healthcare encounters, as well as pregnancy and mental health- related encounters. R61 data will be merged with satellite-derived and modeled estimates of floodwater extents to improve the spatiotemporal characterization of exposures, as well as displacement and outmigration variables. Based on our findings, we will develop and test communication, coordination, and guidance strategies with partners in local health systems and agencies. Results will provide data-driven recommendations for improving the coordination and management of long-term health risks, including mental and maternal health risk during recovery, and identify specific opportunities to enhance flood recovery strategies. We expect that this work will support ongoing recovery efforts in the aftermath of Hurricane Helene and inform future disaster preparedness and flood mitigation strategies in Appalachia and other rural, mountainous areas, contributing to more resilient health systems and communities.

NIH Research Projects · FY 2025 · 2025-09

Project summary/abstract New treatments of neurological disorders are desperately needed, especially in regions with significant and disproportionately unmet needs, such as in Mexico. Although Mexico's incidence of chronic pain is on par with that in the US, their utilization of current treatments only amounts to 0.56% of the level seen in the US. Development of natural products from organism's native to Mexico for the treatment of pain has the potential of fewer perceived risks and may overcome challenges of current therapeutics. The goal of this project is to establish millipedes as a source of natural products that can be leveraged for the treatment of pain. The success that natural products have had clinically is due to their ancient evolutionary history; their structures and functions evolved over millions of years of selection to carry out an essential role for the organism. For example, many of the antibiotics applied today are produced by terrestrial microorganisms that use them to vanquish competitors. Millipede natural products are no different. Different millipedes have evolved the ability to produce unique defensive metabolites, including hydrogen cyanide, oxidized aromatics, and terpenoid alkaloids. These metabolites change in structural complexity through evolutionary time. Their structural diversity is correlated with phylogeny, and the conservation of biosynthetic pathways along with the evolution of gland anatomy to store these metabolites are strong indicators of their important ecological role. The terpenoid alkaloids, nitrogen containing heterocycles, are the least studied but simultaneously the most interesting from a biological perspective. Less than 10% of the alkaloid producing millipedes have been investigated and each one has led to a new chemistry with unprecedented carbon backbones. Most notably, we recently showed that two different subsets of these molecules bind potently and specifically to two different receptors (sigma-1, and sigma- 2) with each receptor having been implicated for the treatment of neurological disorders, including chronic pain. Our research team proposes to study millipedes native to Mexico to investigate the relationship between molecular phylogeny and chemical diversity to uncover novel natural products with important biological activity. Our team's expertise in millipede ecology specific to Mexico, natural products chemistry, and biochemistry, will lead to the discovery of millipedes producing new bioactive alkaloids. Novel natural products will be isolated, structurally characterized, and biochemically evaluated to fully understand their therapeutic potential. Collectively, our proposed research program will broadly impact the field by showing that millipedes native to Mexico are a repository of many new biologically relevant natural products. Studying these small molecules will lead to the discovery of novel sigma-1 and sigma-2 ligands that have the potential to populate the drug pipeline, specifically for the treatment of chronic pain, without the perceived risks of current treatments.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Achilles tendinopathy is a disabling, often painful condition that can progress to further degenerative tissue changes or result in tendon rupture if not effectively managed. It is widely held that therapeutic mechano- transduction processes promote healing of the injured tendon. Clinical ultrasound therapies deliver energy in the form of propagated sound waves to targeted tissues. However, current therapeutic ultrasound treatments commonly provide minimal mechanical stimulation of the injured tendon. Focused ultrasound (FUS) is an emerging, non-invasive therapy capable of inducing bio-effects through mechanical and/or thermal mechanisms with high spatial and temporal precision. Recent pre-clinical tendon models have demonstrated the potential of FUS to achieve mechanical fractionation with no deleterious effects on tendon function or healing; however, no studies have investigated FUS as a rehabilitative loading treatment for tendinopathy. The primary objectives of this study are to (1) utilize numerical modeling to establish FUS protocols for precise acoustic stimulation of murine Achilles tendons, (2) identify FUS stimulation parameters for effective in vivo treatment of murine Achilles tendinopathy, and (3) characterize the cell and matrix responses associated with distinct FUS stimulation regimes. We will first develop an integrated computational modeling - experimental approach to characterize ex vivo tendon temperature changes and mechanical strain under a range of FUS treatment parameters (Aim 1). Protocols developed in Aim 1 will then be applied towards in vivo FUS treatments of injured tendons, with healing assessed using biomechanical, histologic, and cell biologic outcomes (Aim 2). We hypothesize that (a) the application of FUS will augment healing of injured Achilles tendons, and that (b) mechanical FUS regimes will be more effective than thermal FUS regimes. Successful demonstration of the potential of FUS to stimulate tendon healing will considerably strengthen the rationale for using FUS as a non-invasive, rehabilitative treatment method for tendinopathy and provide encouraging evidence of a potentially effective alternative over existing ultrasound therapies for tendon injuries. Furthermore, our project will generate novel insights into the relationships between acoustic stimulation parameters and tendon mechanotransduction. Finally, this project will establish a modular, scalable experimental platform upon which future studies in larger species and/or different tendon types can readily be undertaken.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Glioblastoma is a deadly brain cancer marked by extensive invasion into the surrounding brain. Interstitial fluid flow is heightened during disease development and is a major driver of tumor progression. These fluid flows in and around brain tumors are heterogeneous and can be mapped by using our mathematical modeling approach that uses a single patient or mouse’s dynamic MRI to generate 3D maps of interstitial transport quickly and robustly. Interstitial flow not only changes during tumor progression, but can also change in response to anti- tumor therapy, but the ways in which this happens and the dynamism of these changes are relatively unknown. Here we propose to focus on interstitial fluid flow in glioblastoma to target treatments, assess treatment efficacy, and plan treatment using a range of in vivo and mathematical modeling tools alongside advanced engineering and imaging methodology. First, we will leverage the finding that interstitial fluid flow and transport can be used to identify invading tumor cells to target treatment via focused ultrasound mediated blood brain barrier opening, developing a novel precise FUS system coupled with both validation and treatment to target invasive tumor cells preclinically on demand. Next, we will develop methodology to compare flow fields over the course of a series of standard of care treatments (temozolomide and radiation therapy), physical drug delivery methods (convection enhanced delivery and focused ultrasound), and antibody based targeted therapies (anti-VEGF and anti-PD1). In this way, we will assess the changes to flow acutely and longitudinally and correlate these outcomes with tumor growth and progression. Last, we will leverage advanced computational methods to use initial interstitial flow fields and transport metrics to map drug delivery and disease progression. Founded on our basis of preliminary data we will create physics-based mathematical models to determine the best flow directed therapeutic strategies and will validate our approach with in vivo studies. Overall, this work leverages an untapped biophysical biomarker, interstitial fluid flow, in glioblastoma to target treatment, understand treatment responses, and plan treatment. With this new understanding of the dynamics and significance of interstitial fluid flow in treatment of GBM, we can translate our methodologies and outcomes to patients.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY / ABSTRACT As children’s media consumption shifts to platforms like YouTube, young children are increasingly exposed to rapid video-switching, a feature that may disrupt attention and learning. This developmental cognitive neuroscience study will investigate how video-switching affects young children’s attention and comprehension. Using a within-subject experimental design, we will manipulate two viewing conditions—switching and non- switching—on a custom-built platform that simulates YouTube. Three-year-old children will watch educational videos while their sustained and selective attention are measured at both neural and behavioral levels using EEG and eye-tracking. After viewing, we will assess children’s comprehension. Additionally, we will analyze switching behaviors in both naturalistic and controlled settings to test potential associations. Our study addresses a significant gap in understanding how media design features, specifically video-switching, impact early cognitive development. Given the urgent need to evaluate the effects of these features on young children, our research will provide critical evidence to inform guidelines for healthy media use in early childhood. Specifically, we will address three aims: 1) assess the impact of video-switching on attention, 2) examine its effects on comprehension, and 3) investigates associations between switching behaviors in naturalistic and experimental settings.

NIH Research Projects · FY 2025 · 2025-09

Modified Project Summary/Abstract Section Aedes aegypti transmits several arboviral diseases, including dengue and Zika fever, which threaten virtually half of the world’s human population. Two subspecies, Ae. aegypti aegypti (Aaa) and Ae. aegypti formosus (Aaf), have been described based on their body coloration. These subspecies differ remarkably from each other in their worldwide distribution, association with humans, and ability to transmit pathogens. In Anopheles populations, polymorphic inversions are often responsible for epidemiologically important phenotypes but our knowledge about chromosomal rearrangements in Aedes populations is limited. Although our recent study identifies a large pool of structural variation in the Ae. aegypti genome, their impact on behavior and adaptations of this species remains completely unknown. We hypothesize that chromosomal inversions contribute to the remarkable phenotypic and genetic plasticity of Ae. aegypti. However, more studies are needed to better understand the geographical patterns of the inversion distribution all over the globe and their potential involvement in ecotypic differentiations of the Ae. aegypti subspecies. Our project will test whether chromosomal rearrangements are 1) subspecies specific; 2) associated with ecological variables in Africa; 3) involved in mosquito adaptations to human. In this study, we will take advantage of the improved, fully re-annotated genome assembly for Ae. aegypti, employ the Hi-C approach and SNP-based inversion genotyping in publicly available WGS sequencing data to characterize chromosomal rearrangements across the world-wide distribution. We will also apply simple and robust PCR-based approaches to identify presence of common African inversions in individual mosquitoes in Senegal and use RNA-seq to determine differences in transcriptional profiles of standard and inverted genotypes. Our long-term goal is to understand the underlying genomic determinants of epidemiologically important phenotypic and behavioral differentiation of Ae. aegypti mosquitoes. Toward this end, we propose three specific aims: 1) identify patterns of world-wide distribution of the chromosomal rearrangements in Ae. aegypti and their associations with Ae. aegypti subspecies; 2) test if widely spread chromosomal inversions are associated with ecological variables in Africa; 3) determine specific transcriptional profiles associated with standard and inverted homokaryotypes of common African inversion 1pA. The discovery of chromosomal inversions in Ae. aegypti will stimulate future genetic studies aimed at preventing mosquito-borne disease transmission

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Exercise is a powerful strategy to improve skeletal muscle metabolism that can both prevent and reverse disease. However, the complex signaling that drives the immediate and long-term changes in muscle metabolism is incompletely understood. Skeletal muscle consists of multiple cell types, such as myocytes, vascular endothelial cells, pericytes, and immune cells. When subjected to exercise stress, these cells communicate with one another to coordinate the heightened energy production required by the skeletal muscle. Our long-term goal is to understand intercellular skeletal muscle signaling initiated in response to exercise, thus informing strategies to promote health and prevent disease. Reactive oxygen species (ROS) are molecules known for their ability to both initiate signaling cascades and to cause damage. Skeletal muscle produces ROS in response to exercise, but whether ROS harm or protect the muscle has been debated. Initially, ROS were believed to be solely deleterious, contributing to conditions like diabetes and sarcopenia. However, recent findings indicate that ROS play a crucial role in skeletal muscle's metabolic adaptation to exercise. Despite this, we still have limited knowledge about where ROS are produced, the enzymes involved, and the specific processes that rely on ROS for beneficial signaling during exercise. Building on our previous research, our current proposal focuses on NADPH oxidase 4 (NOX4), a ROS-producing enzyme we identified as a critical factor in initiating skeletal muscle's metabolic responses to exercise. Importantly, this enzyme is most highly expressed in the vascular endothelial cells of skeletal muscle. If we remove this enzyme from only the endothelium, we observe a reduced metabolic response to acute exercise. Based on these findings, we will investigate the hypothesis that endothelial signaling, dependent on NOX4, plays a crucial role in determining the mitochondrial metabolic responses to exercise in skeletal muscle. To investigate this, we will utilize advanced tools that allow us to specifically manipulate the expression of Nox4 in endothelial cells. This will enable us to gain new insights into exercise-induced endothelial signaling and examine the impact of endothelial cell-derived ROS on both 1) endothelial signaling and 2) the mitochondrial responses to exercise in skeletal muscle. Ultimately, we hope these findings will contribute to developing targeted exercise interventions and potentially provide a foundation for treatment strategies based on exercise as medicine.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Venezuelan equine encephalitis virus (VEEV) can cause significant morbidity including neurocognitive impairment, resulting in a significant impact on population health, healthcare costs, and workforce productivity. VEEV is an important human and veterinary pathogen belonging to one of seven antigenic complexes in the genus Alphavirus, family Togaviridae. Its importance is recognized by multiple government agencies, with VEEV being classified as a category B priority pathogen by NIAID and as a select agent by both the CDC and the USDA. NIAID has also listed VEEV as a prototype pathogen for the Togaviridae family, indicating it should be utilized for medical countermeasure development for other Togaviruses. Neurological cases of VEEV have a mortality rate as high as 35% in children and 10% in adults, with long-term neurological deficits observed in 4-14% of survivors. Sequelae observed include convulsions, seizures, paralysis, confusion, photophobia, intellectual disability, emotional instability, and behavioral changes. There are currently no FDA-approved antiviral therapeutics for the treatment of VEEV. However, there has been significant effort on the development of small molecule compounds targeting encephalitic alphavirus infections, with a few candidates showing efficacy in animal models. Of these, ML-336 and its derivatives (BDGR compounds) have shown great promise in mouse models of encephalitic alphavirus infections. BDGR-49, a new chemotype, has shown prophylactic efficacy against VEEV and EEEV and 100% therapeutic efficacy against VEEV in mouse models. A critical gap in knowledge is whether BDGR-49 can prevent VEEV induced long-term neurological sequelae when administered therapeutically. We hypothesize that BDGR-49 treatment will result in reduced VEEV replication and neuroinflammation allowing prevention of long-term neurological sequelae. To address this hypothesis, we propose the use of a well characterized mouse model of VEEV TC-83 infection which recapitulates neurological sequelae observed in humans infected with VEEV and an antiviral, BDGR-49, which has also been extensively characterized in VEEV-infected mice. Specifically, mice infected with VEEV TC- 83 display deficits in prepulse inhibition (the inability to filter out extraneous sensory stimuli), thalamus damage (glial nodules and calcification), and glial activation in the posterior nuclear complex of the thalamus and the hippocampus dentate gyrus, a central hub for cognitive function. We have preliminary data showing that mice have anxiety and memory deficits, and multiple neuropathological markers including gliosis, perivascular cuffing, and neuroinflammation and cell death, and neuronal loss within the dentate gyrus up to 90 days post-infection. Collectively, these preliminary studies established the utility of this model for studying neuroinflammation and cognitive impacts following VEEV infection. This model will be used to test our hypothesis in two specific aims: Aim 1: Determine the impacts of prophylactic BDGR-49 treatment on VEEV induced neurological sequelae and Aim 2: Determine the impacts of therapeutic BDGR-49 treatment on VEEV induced neurological sequelae.

NIH Research Projects · FY 2025 · 2025-08

Cigarette smoking causes more than a dozen cancers, including cancers of the head and neck, colon, bladder, and lung. Although quitting smoking drastically reduces the risk of cancer mortality, rates of cessation are low, particularly in rural areas. Current evidence-based treatments for smoking cessation have made progress, but substantial room for innovation remains and novel strategies are needed. Successful smoking cessation may be considered, in part, an intertemporal choice between continuing to smoke and achieving the delayed health benefits associated with quitting (e.g., avoidance of lung cancer). Thus, quitting requires one’s behavior to be sufficiently motivated by future outcomes. Unfortunately, robust cross-sectional and longitudinal evidence indicates that people who smoke cigarettes show elevated delay discounting, or a bias for immediate gratification. Our work and others’ show that delay discounting predicts treatment failure and relapse in smoking cessation. These findings suggest that delay discounting often prevents successful quitting because the health benefits of cessation (e.g., avoiding cancer) are too delayed to motivate sustained behavioral change. Thus, delay discounting is a therapeutic target in smoking cessation, where interventions that increase valuation of future outcomes may facilitate quitting. Episodic future thinking (EFT) is one such scalable intervention that is designed to shift time perspective and reduce bias for immediate gratification by promoting vivid and frequent visualization of a broad range of personally significant future events (e.g., weddings, birthdays, spending time with loved ones). In laboratory studies, we have shown that this form of generalized EFT in cigarette smokers reduces delay discounting and motivation to smoke (i.e., cigarette consumption, craving, and valuation). Moreover, we have strong preliminary evidence that cancer-related EFT, a novel form of the intervention that involves visualizing the future experience of smoking-related lung cancer, may be more efficacious for reducing urges to smoke than the generalized form. In the proposed trial, we will adapt both forms of EFT (generalized and cancer-related) for clinical use in smoking cessation. Remote intervention delivery and outcomes assessment (breath carbon monoxide and number of cigarettes smoked per day) will be used to minimize clinical burden and increase intervention reach and access, including in rural populations who often lack access to evidence-based treatments for smoking cessation. In an eight-week proof-of-concept trial, participants will be prompted to engage in EFT or control episodic thinking multiple times per day and during acute craving episodes. Specific Aims 1 and 2 will examine the feasibility and efficacy, respectively, of generalized and cancer-related EFT to reduce smoking. An Exploratory Aim will investigate the potential moderating role of income, education, sex, and other characteristics (e.g., baseline DD, cigarettes/day) in the effects of EFT on smoking outcomes.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Protein lysine methylation is a post-translational modification (PTM) that neutralizes the positive charged side chain and modulates protein functions in many key biological events. In metazoans, somatic cells displaying distinct cellular phenotypes are originated from one zygote and contain the identical sequence of DNA. Upon differentiation, transition of cell fate is initiated by lineage transcription factors and a cell identity is stabilized by epigenetic factors that define a chromatin state. Distinct histone lysine methylation is central to active or repressive chromatin formation. Dysregulation of this process often leads to pathogenic conditions such as developmental disorders or cancer. Once an epigenetic state is established, cells need to faithfully recapitulate the steady state in mitosis in order to safeguard cell identity and prevent developmental dysregulation and oncogenic transformation. The epigenetic inheritance is a new research discipline undergoing rigorous investigations. Despite that significant advancement has been achieved to decipher the mechanisms of repressive chromatin inheritance, how active chromatin is inherited, involving temporo-spatial regulations of histone lysine methyltransferases, still remains elusive. In addition to histones, histone lysine methyltransferases also catalyze non-histone methylation events, whose importance is emerging in the context of early development. However, there is a lack of systematic investigation of these non-canonical substrates and how they critically regulate distinct biological processes. In this proposal, we will address these key knowledge gaps by interrogating the mechanistic questions in active chromatin inheritance and develop a new approach to investigate non-canonical methylation events by histone lysine methyltransferases. We expect to establish new research platforms and advance our understanding towards methylation biology.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY The proposed work will build on current wearable, optically pumped magnetometer-based magnetoencephalography (OPM-MEG) methods our group has developed for use in infants, and will further optimize this technology for simultaneous recording of brain activity in caregiver-infant dyads during naturalistic, freely moving, social interactions. While both electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) have been used to assess brain function in infants, and EEG has been used to simultaneously assess caregiver-infant dyadic brain function during social interactions, they each have limitations that are addressed by wearable OPM-MEG. Both EEG and MEG allow for high temporal, millisecond, resolution; however, EEG poses challenges for source localization. Although fMRI addresses the source localization issues of EEG, it does not allow for the same naturalistic social paradigms as EEG, and is also loud and expensive. OPM-MEG is non-invasive, quiet, and provides both high temporal resolution and accurate source localization. Despite the strengths of OPMs, there are several outstanding challenges that include 1) optimization of source imaging in caregiver-infant dyadic assessments, as well as 2) developing robust, motion-tolerant OPM-MEG approaches for ambulatory hyperscanning. This proposal will address all of these challenges in caretaker-infant dyads with 3-6-month-old infants, a group where unplanned movements are inevitable. Addressing these issues in this population and paradigm will answer incredibly important technological questions in a context that will translate to many populations where functional neuroimaging is needed to optimize diagnosis and treatment. Further, the ability to assess brain function simultaneously within a dyad with lifespan compliance represents an unprecedented opportunity to gain knowledge about social neuroscience, which is key to being able to recognize clinically relevant patterns of brain function indicative of later emerging and current psychopathology. Once the proposed work is completed, we will have developed optimized protocols, and the highest quality OPM- MEG data ever collected in infants, as well as simultaneous caregiver-infant OPM-MEG collected during naturalistic, physically-touching, social interactions. The innovations represented by this proposal are paramount to moving the field of functional neuroimaging vertically to improve our understanding of neurodevelopment and brain function using salient social interactions and paradigms requiring motion.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY: Despite aggressive definitive treatment of osteosarcoma (OS), survival expectations for human and canine OS have not improved in decades due to refractory metastatic disease. The poor immune responsiveness of osteosarcoma to immunomodulating therapies contributes to the persistence of metastatic disease development. Novel therapeutic paradigms to activate this “cold” tumor microenvironment and induce an effective anti-tumor immune response in OS are direly needed. To fulfill this need, our research team proposes a combination therapy approach using 2 highly novel modalities – histotripsy tumor ablation and N-dihydrogalactochitosan (GC) to treat OS. Histotripsy is a non-thermal, non- invasive and non-ionizing tumor ablation technique that utilizes high intensity focused ultrasound waves to mechanically disintegrate tissue. Histotripsy releases non heat-denatured tumor antigens, which can potentially induce a more robust anti-tumor immune response. GC is a semisynthetic derivative of chitin with demonstrated ability as an immunostimulant that enhances the anti-tumor effects of various ablation modalities across different cancer types. Despite the exciting potential of histotripsy and GC to synergistically induce an effective anti-tumor response, combining both therapies to treat OS has not been reported. Thus, this multidisciplinary proposal will evaluate combining histotripsy tumor ablation with GC as an innovative immunomodulatory strategy for treating osteosarcoma using a unique comparative oncology study that incorporates murine preclinical models and pet dogs with spontaneous appendicular osteosarcoma. The overall objective of our proposed study is to evaluate local and systemic immune responses in OS after combining histotripsy tumor ablation with intratumoral injection of GC. Our overall hypothesis is histotripsy-GC treatment of OS induces effective and durable intratumoral and systemic anti- tumor immune responses. We will test this hypothesis with the following aims: Aim 1: Characterize the immunomodulatory responses to and effects on metastasis of histotripsy-GC treatment of OS in a murine heterotopic syngeneic tumor model. We hypothesize that histotripsy-GC treatment of OS heightens the intratumoral and systemic immune activation profiles, induces an abscopal effect, and reduces metastatic burden. Aim 2: Evaluate the feasibility of histotripsy-GC treatment of OS in dogs with appendicular OS, and the effects of treatment on the immune profile of the OS TME and on progression-free survival. We hypothesize that histotripsy-GC treatment of canine appendicular OS does not result in major adverse events, stimulates increased intratumoral immune cell infiltration, upregulation of intratumoral and lymph node immune activation gene profiles, and achieves increased PFS compared to standard-of-care therapy. Significance: These data will inform subsequent studies that generate clinically relevant data to design future human clinical trials for histotripsy-GC therapy in OS.