Washington University

NSF Awards · FY 2026 · 2026-10

Large Language Models (LLMs) have shown impressive performance on pure reasoning tasks like math questions and logic puzzles. However, Artificial Intelligent (AI) systems with such LLMs still struggle with complex tasks that require finding and combining information from multiple external sources. For instance, answering a complicated legal or scientific question often requires a step-by-step investigation where the answer to one question determines what to search for next. While current AI systems can perform multiple searches, they typically rely on a single, general-purpose model that struggles to form a structured plan or adapt its expertise to different phases when investigating a problem in depth. This project addresses this critical gap by creating a new way for AI systems to actively seek out and synthesize knowledge. By empowering models to formulate step-by-step research plans and adapt to each part of a problem, this project promotes the progress of science and advances national prosperity through the creation of highly reliable and transparent decision-making tools. These new capabilities will directly benefit evidence-based fields such as legal analysis and scientific discovery. In addition to these technological benefits, the project supports education by creating new university courses that teach students how to critically analyze AI systems. The investigator will also lead hands-on outreach activities for local school students to inspire the next generation of researchers. The technical goal of this award is to establish a proactive, efficient, and transparent reasoning framework for knowledge-intensive tasks. The research activities are organized into three integrated thrusts. The first thrust develops the Adaptive Knowledge Synthesis framework, which trains LLMs to formulate structured reasoning plans through hypothesis-driven decomposition. Within this framework, a conductor model dynamically reconfigures a base LLM to create specialized experts tailored to specific sub-tasks, using confidence-guided routing to synthesize a final answer. Because this multi-step process generates long dialogue histories and extensive intermediate outputs, the second thrust develops scalable inference methods to reduce computational costs. On the input side, an activation-level integration will be introduced to effectively process long contexts. On the output side, the research team will develop compressed reasoning techniques and sample-efficient test-time scaling methods to streamline generation. Finally, the third thrust establishes a new generation of evaluation frameworks to rigorously measure both procedural correctness and computational efficiency. This involves building controllable benchmarks with systematically varying difficulty, robustness tests featuring multi-hop contradictions, and efficiency-aware metrics that evaluate the trade-off between reasoning accuracy and computational cost. All resulting models, algorithms, and evaluation frameworks will be released as open-source resources to benefit the broader research community. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2026 · 2026-08

Poisson 2026 Summer School and Conference will be held at the University of Antwerp and KU Leuven in Belgium from August 3-14, 2026 . This award will provide partial support for U.S.-based participants to attend this international event, which focuses on Poisson geometry, a mathematical framework connecting ideas from geometry, algebra, and mathematical physics. By combining an introductory summer school with a research conference, the project lowers barriers to entry into a highly active area of mathematics while helping to build a diverse and well-connected international research community. Broader impacts include advanced training for graduate students and postdoctoral researchers, expanded international engagement for U.S.-based mathematicians, a public lecture accessible to general audiences, and freely available online recordings that will extend the reach of the program worldwide. The scientific program has two complementary components. The summer school will feature five mini-courses introducing core topics and recent developments in Poisson geometry and related areas, aimed primarily at graduate students and early-career researchers. The conference will include approximately twenty plenary lectures by leading researchers presenting current directions, recent advances, and open problems. The program emphasizes interactions among Poisson geometry, symplectic geometry, Lie theory, noncommutative geometry, and mathematical physics. It also prioritizes support for early-career U.S. participants and broad international visibility, with a strong emphasis on mentoring, networking, and dissemination through recorded lectures and a planned special journal issue. Additional information is available at: https://wis.kuleuven.be/events/2026poisson/poisson2026 This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2026 · 2026-08

Tendons are essential tissues that connect muscles to bones and transfer forces to enable movement. Instead of being uniform mechanical links, tendon properties can vary significantly across individuals and even within a single tendon. These differences in local properties may play a key role in determining how tendons function, adapt to physical activity, and respond to injury. However, most studies simplify tendons as uniform materials, overlooking potentially important variations that could influence health and healing. This research project will reveal how local changes in tissue properties affect tendon performance and cellular behavior. By improving understanding of how tendons function, this research will provide valuable information to guide translational efforts aimed at improving injury prevention, refining rehabilitation strategies, and inspiring new regenerative therapies. This study will also integrate research with education by developing new course materials on tissue heterogeneity, training interdisciplinary teams of student scientists, and engaging middle and high school students through interactive outreach activities that highlight the value of teamwork and interdisciplinary science. This project will determine how local tissue properties and spatial heterogeneity drive tendon mechanics and mechanobiology. Full-field maps of morphological, microstructural, compositional, and mechanical properties will be determined in tendons with differing levels of inherent heterogeneity. Spatial statistical approaches, including autocorrelation metrics and cross-correlation analyses, will quantify heterogeneity and identify relationships between local tissue properties and mechanical function. Multivariate regression models will determine which spatial features best predict local- and tissue-level mechanics. To investigate how spatial heterogeneity influences tendon mechanobiology, cellular responses to load will be measured in calcium-tagged mouse tendons then correlated with local properties and heterogeneity metrics. Comparisons between native and recellularized tissues will isolate matrix-driven effects, and regression models of multi-modal spatial data will determine best predictors of active responses to load. By integrating high-resolution mapping with advanced spatial analysis, this work will establish a new framework for linking tissue heterogeneity to mechanical and mechanobiological responses of biological tissues. Results will guide translational interventions to prevent tendon injury and inspire novel biotechnology advances to enhance treatment strategies. In addition, these innovative techniques could enable transformative advances in understanding the mechanics and mechanobiology of other soft tissues, enhancing overall impact. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2026 · 2026-07

Modern society depends on computer networks for daily life, economic activity, and essential public services. Yet these networks remain difficult for people to manage safely and reliably because many operational tasks are detailed, repetitive, time-sensitive, and easy to perform incorrectly. This project advances a new research vision called Technological Management of Computer Networks: network systems should be designed not only to carry data, but also to make their own operation understandable, governable, and trustworthy. In this vision, people retain responsibility for judgment, policy, and accountability, while artificial intelligence agents carry out low-level operational tasks within approved limits. The central research challenge is to determine how such delegation can be made safe and robust: automated actions must be tied to human authority, checked against policy, controlled within defined boundaries, recorded as evidence, and open to review. By treating management, operations, and compliance as core technical properties of network systems rather than as external paperwork or after-the-fact automation, the project aims to create a new foundation for reliable and accountable digital infrastructure. The work will support national needs for secure networks by reducing preventable failures, improving trust in AI-assisted operations, and training students in secure, controllable, and robust network systems. This project will develop and evaluate the scientific and technical principles needed to make technological management practical. The research will formalize a human and artificial intelligence division of responsibility in which people provide governance and accountability by defining policies, approving procedures, assigning authority, evaluating tradeoffs, and reviewing evidence. Artificial intelligence agents provide fulfillment by interpreting operational intent, inspecting telemetry, selecting approved workflows, preparing configuration changes, and carrying out routine tasks within technical and organizational limits. The project will use both frontier artificial intelligence models, which provide leading capabilities for reasoning and planning, and locally hosted models, which can support privacy, cost control, availability, and operation close to managed systems. The project will investigate how signed workflows, role-based authority, policy-controlled system components, tamper-resistant operational logbooks, and improving artificial intelligence capabilities can be combined so that network operations are not merely automated, but verifiable, auditable, and accountable. The work will be built and tested using an open-source, HTTP-based overlay networking platform developed from foundational open-source technologies, including Linux, Envoy Proxy, and the Open Policy Agent. The results are expected to be useful for managing these technologies individually, as well as for managing them together as a policy-based overlay network. Experiments will measure safety, responsiveness, governance, and compliance evidence compared with more manual forms of network management. Example tasks include detecting latency problems, adjusting network parameters within approved limits, carrying out staged configuration rollouts, reviewing access permissions, rotating keys and certificates, and preparing evidence for audits. The project will produce open-source software, repeatable benchmark experiments, and a capstone remote-access overlay system that demonstrates a new model of artificial intelligence-assisted network operations under human governance. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2026 · 2026-07

Mathematics advances when seemingly different phenomena turn out to share a common underlying structure. This project investigates how mathematical constraints shape the behavior of mathematical objects across several interconnected areas of pure and applied mathematics. A central theme is rigidity: when does satisfying a natural condition force an object to have a very special form? Discovering such principles deepens basic scientific understanding and creates tools that can be used in other fields. The project serves the national interest by advancing the mathematical sciences, training doctoral students and undergraduates, supporting public outreach, and developing ideas with connections to quantum information, machine learning, and biomedical science. In particular, the applied part of the project seeks better mathematical models of disease progression in Alzheimer's disease and multiple sclerosis, with the long-term aim of improving the design and interpretation of clinical trials. The project develops this theme in four connected directions. The first concerns the complex geometry of the polydisk: the goal is to classify subvarieties whose intrinsic Caratheodory metric agrees with the one inherited from the ambient domain, and to determine how this geometric condition relates to holomorphic extension and Pick interpolation. The second develops a unified theory of complete Pick spaces, a broad class of function spaces that includes the Hardy space as a prototype. The investigator studies factorization, multiplier algebras, Hardy-type function scales, and corona-type problems in this general setting, with the aim of explaining why many classical Hardy space theorems extend far beyond their original context. The third extends classical random matrix theory to tuples of matrices satisfying algebraic constraints, particularly commuting tuples. This opens a new direction with natural connections to quantum information, where commutativity encodes the compatibility of measurements. The fourth applies mathematical analysis to scientific problems, especially the development of improved two-parameter models of disease progression in Alzheimer's disease and multiple sclerosis using brain age derived from MRI data. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT This project aims to investigate the roles of a newly identified population of neurons in the supramammillary nucleus (SuM) that are essential for threat learning and avoidance behaviors. These neurons are activated by diverse stressors and may play critical roles in threat conditioning and persistent avoidance. The specific aims of this research are to: (1) elucidate the temporal dynamics and causal roles of these neurons during threat learning, avoidance, and extinction using fiber photometry, optogenetics, and behavioral assays; (2) determine whether these neurons selectively excite neuron populations in the nucleus accumbens (NAc) to promote threat avoidance; and (3) reveal the neural types in SuM and which are recruited during threat learning and sustained avoidance using advanced molecular techniques such as Projection-TAGs, barcoded retrograde viruses, and Act-seq with single nucleus RNA sequencing. By characterizing this novel neural circuitry, the project aims to advance our understanding of active coping mechanisms and contribute to the development of new treatments for stress and anxiety-related disorders by identifying potential therapeutic targets.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Major traumatic injury, particularly when accompanied by hemorrhagic shock (HS), results in disruption of endothelial barrier integrity, leading to vascular leakage and tissue edema. Patients with traumatic brain injury (TBI) are especially vulnerable to HS, as a two-fold increase in mortality is observed when TBI is accompanied by HS. The proton sensing G protein-coupled receptors (PsGPCRs), GPR4 and GPR68, are expressed on cerebral endothelial cells and are regulated by extracellular pH. These receptors play a critical role in maintaining endothelial barrier function and may be pathologically activated in the acidotic state generated during HS. We have recently established a murine model of combined TBI and HS that closely recapitulates human injury. Preliminary data from this model demonstrates increased endothelial permeability when HS is introduced. Additionally, we have observed upregulation of cerebral endothelial PsGPCR expression following murine TBI, suggesting a potential role for these receptors in mediating endothelial activation and dysfunction in response to injury. Central Hypothesis: PsGPCRs mediate endothelial activation and increased cerebral permeability following TBI, and their activation is exacerbated by the systemic acidosis resulting from HS, leading to worsened cerebral edema and neurobehavioral outcomes. To test this hypothesis, we propose three specific aims: 1. Investigate how trauma and hemorrhagic shock influence the expression of cerebral endothelial PsGPCRs. 2. Identify how PsGPCR activation, induced by trauma and hemorrhagic shock, affects cerebral endothelial cell permeability and signaling. 3. Assess the impact of PsGPCR inhibition on neurobehavioral deficits following HS+TBI This work will fill a critical gap in our understanding of the molecular mechanisms underlying endothelial barrier dysfunction after traumatic injury and hemorrhagic shock. The results of this study may identify PsGPCRs as novel therapeutic targets for limiting cerebral edema and improving neurobehavioral outcomes for those suffering with TBI.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY La Crosse virus (LACV) is the leading cause of pediatric arboviral encephalitis in the U.S., causing severe neurological consequences and economic burdens. Understanding LACV entry mechanisms is essential, as this knowledge will aid in the development of targeted antiviral therapies. The proposed research plans to elucidate the molecular mechanisms underlying La Crosse virus entry via a newly identified host factor, Jagged2 (JAG2). Aim 1 of this proposal seeks to determine the role of JAG2 and its homologs in LACV glycoprotein-mediated entry (Aim 1). The role of JAG2 as a cellular receptor for LACV entry will be investigated using cutting-edge genetic, biochemical, and live imaging approaches. Investigate the impact of JAG2 homologs from insect and rodent reservoirs on LACV infection. Aim 2 will delineate key interactions between LACV glycoproteins and JAG2, showing that JAG2 epidermal growth factor (EGF)-like domains bind directly to LACV glycoprotein Gc. Deep mutational scanning, neutralization assays, and biolayer interferometry (BLI) will be used to identify interaction sites. Infectious LACV Gc mutants will map viral determinants of JAG2 neutralization. This study will define the role of JAG2 in LACV infection, uncover new viral entry mechanisms, and lay the groundwork for future in vivo studies in both the mammalian host and the insect vector. These findings could contribute to broad- spectrum antiviral strategies targeting similar viral pathogens. The proposed research plan will be complemented by a personalized and comprehensive training plan to ensure that Jana Liese develops a deep understanding of cellular biology, virology, and biochemistry as well as technical and leadership skills essential for a career as a physician-scientist. This training will be further enriched by active participation in a highly supportive and collaborative interdisciplinary research community at Washington University in St. Louis School of Medicine.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT We propose to broadly disseminate and extend intuitive, powerful cloud-based resources for optical brain mapping that facilitate efficient, accurate, and standardized processing that will harmonize the emerging set of optical measurement strategies within the growing ecosystem of network level analyses used throughout the greater brain mapping community. The neuroimaging community faces numerous challenges in data collection, preprocessing, estimation of brain connectivity, and analyses of relationships between brain connectivity and behavior. An ever-expanding community of researchers are employing optical methods based on functional near infrared spectroscopy (fNIRS) in order to infer pathophysiological states of the brain for detection/ characterization of disease or cerebral hemodynamics for understanding human brain health, development, and aging. Recent developments of high-density diffuse optical tomography (HD-DOT), a silent, flexible, and scalable technology have demonstrated dramatically improved anatomical specificity and image quality over traditional fNIRS. Further, recent developments in wearable HD-DOT, even using frequency domain and time resolved strategies, open the door to unconstrained mapping of naturalistic human brain function with superior image quality than previously possible. Given the growing worldwide adoption of fNIRS and HD-DOT methods and further developments of next-generation optical brain mapping methods via the BRAIN Initiative, there is an urgent and present need for standardized, accessible and flexible tools that directly support workflows from optical tissue parameter recovery to functional brain mapping to relating variance in brain function to behavior and outcome. Our team is funded by U24NS136402 to address these needs in the optical brain mapping community by developing and validating computational tools including NIRFASTer, NeuroDOT, and Network Level Analyses (NLA), for tissue parameter recovery, optical brain mapping, and model-based connectome-wide association studies of brain function and behavior, respectively. We address the unmet needs for data resources with: (1) greater dissemination and training for our tools with detailed documentation, training materials, and international workshops, (2) cloud deployment of our software to increase scale and accessibility, while easing the computational burden for the user, and (3) expanded utility of these sustainable, flexible tools to meet the evolving needs of users at the forefront of optical imaging technology development. This R50 will support a Research Software Engineer (RSE) to be an established senior RSE through gaining deeper experience expanding development, maintenance, and dissemination of the U24-supported tools while enhancing her skills and knowledge of best software engineering practices, FAIR standards, and optical neuroimaging methods, as well as common and emerging neuroimaging algorithms. This proposal directly supports execution of BRAIN Initiative goal 7 that seeks to integrate new technological and conceptual approaches to discover how dynamic patterns of neural activity are transformed into cognition, emotion, perception, and action in health and disease.

NIH Research Projects · FY 2026 · 2026-06

Abstract The long-term goal of this research program is to understand and identify mechanosensitive mechanisms that regulate cellular transcription and cell-to-cell signaling during normal tissue morphogenesis and patterning. For instance, our sense of touch, regulation of blood pressure, osmotic regulation, and sense of balance are all regulated by mechanosensitive proteins throughout the body. Our particular interest is in the role that mechanosensitive ion channels and structures, such as the primary cilia, play in the transfer of electrical currents, ions, and second messengers, to promote transcriptional changes and altered cell-to-cell communication. Deciphering these changes is critical for understanding how cellular communication within a complex tissue environment is coordinated and maintained. The importance of mechanosensitive signaling is underscored by the association of many disease states with compromised mechanotransduction, including congenital heart defects, muscle degeneration, arrhythmias, polycystic kidney disease, and numerous neuronal disorders. Despite this, a relatively small amount is known at the level of normal, healthy tissues about how mechanosensitive proteins go from sensing force to eliciting changes in cellular signaling and/or function. Our interest therefore lies in understanding how cells assimilate ‘data’ from mechanosensitive inputs to alter transcriptional programs and cell-to-cell communication to coordinate individual cellular movements within tissues. To carry out this work, we capitalize on our strengths in studying tissue patterning in the zebrafish along with sophisticated 3-dimensional in vitro tissue modeling assays where we can tune and control mechanical forces. From our work we aim to understand: 1) how mechanotransduction affects intracellular signaling, particularly though the activation of transcriptional networks and altered gene expression, and 2) how mechanosensation affects intercellular signaling activities to alter patterning of tissues. We target and utilize highly mechanosensitive cells, such as endothelial cells, smooth muscle cells, and epidermal cells, for our studies to understand both generalizable and cell type specific roles of mechanotransduction in regulating gene expression, cellular motility, and cell-to-cell communication. With this award, we will continue to carry out cutting-edge work aimed at unravelling the role of mechanosensitive proteins and their regulation of cellular differentiation, transcriptional programs, and tissue patterning events.

NIH Research Projects · FY 2026 · 2026-06

SUMMARY Picobirnaviruses (PBVs) are small double-stranded, bisegmented RNA viruses that are found in humans, other mammals and birds. PBVs are among the most commonly detected RNA viruses in the human enteric tract, and they have been linked to human diseases. For example, PBV is associated with type 1 diabetes and detection of PBVs is predictive of the development of severe graft-versus-host-disease in hematopoietic stem cell transplant recipients. These observations raise a key question as to whether PBVs play causal roles in these diseases. However, since no PBV culture or animal model exists, it is currently impossible to experimentally test disease causality. The lack of any PBV isolate is the rate-limiting step in further characterization of the basic biology of PBVs and their role in pathogenic outcomes. Key to overcoming this technical barrier is the understanding of the type of host organism(s) PBVs infect. Dogma asserts that PBVs are human and animal- infecting viruses, but definitive proof of this is lacking. Instead, recent studies support the hypothesis that PBVs are RNA phages that infect bacteria. For example, there is high prevalence of bacterial ribosome binding sites (Shine-Dalgarno) preceding PBV ORFs, a feature characteristic of most phages. In addition, many PBVs encode proteins that can lyse bacteria (lysins) a property necessary for phages to egress from their host bacteria. Critically, in preliminary data we demonstrate that PBV3 can be cultured anaerobically in a stool-derived bacterial community, establishing the first in vitro culture for any PBV. Furthermore, some antibiotic treatments completely block PBV3 growth, and PBV3 RNA and conserved bacterial 16S rRNA co-localize in the same bacteria from in vitro cultures. To identify the hosts of PBVs, we will optimize a bacterial single-cell RNA-sequencing (scRNAseq) approach for co-detection of phage and host RNA in individual bacteria from complex communities. In parallel, specific PBV-targeted FISH- and antibody-FACS approaches will be used to purify and identify PBV infected bacteria. As a key step towards obtaining pure PBV isolates, we will harness germ-free mice to propagate PBVs in vivo. As shown in our preliminary data, germ-free mice gavaged with PBV-containing human stool specimens serve as a vessel to propagate PBVs in vivo. This innovative approach will generate renewable quantities of infectious PBVs and their host for in vitro isolation efforts. Guided by this information and results from scRNAseq, antibody-, and FISH-based assays, we will isolate PBVs by infecting bacterial monocultures of the candidate hosts. Importantly, lytic RNA phage proteins have been coined “protein antibiotics”. PBV-encoded bacterial lysins provide a unique opportunity to characterize new lytic phage proteins and mechanisms that could provide an alternative to traditional antibiotics for treatment of bacterial infections. The overall goals are to: (1) Identify the bacterial hosts of PBVs; (2) establish PBV culture systems in vivo and isolate PBVs in vitro; (3) Define mechanisms of action of PBV-encoded bacterial lysins.

NIH Research Projects · FY 2026 · 2026-06

SUMMARY According to the World Health Organization, air pollution leads to 7 million premature deaths annually, ac- counting for 11% of global mortality. In the United States, 400-1,100 disability-adjusted life-years (DALY) per 100,000 people are lost due to indoor air pollutant exposure. Given that humans spend about 90% of their time inside, our primary chemical exposures occur indoors, whether from indoor activities or outdoor pollutants infil- trating buildings. Indoor dust is a well-studied chemical reservoir containing house dust mites (HDM), skin cells, pollen, soil, clothing fibers, hair, pet dander, and absorbed chemicals, making it a complex entity. HDM and their proteins are known allergens within house dust that can trigger allergic inflammatory responses in humans. However, there is limited understanding of how chemical alterations from reactive indoor pollutants affect HDM and HDM-sorbed chemicals, and subsequently, human health. Our recent research has indicated that exposure of HDM to ozone (a common indoor reactive gas and outdoor air pollutant) and diesel fuel chem- icals (found in homes near roadways where diesel exhaust is present) leads to increased oxidative potential of HDM. Using a mouse model of allergic airway inflammation (AAI), akin to human asthma, we demonstrated that chemically altered HDM exacerbates inflammation, correlating with increased production of allergic cyto- kines. Our central hypothesis is that chemical modifications to HDM from environmental pollutant ex- posure enhance their oxidative potential, resulting in more severe inflammation in an HDM model of AAI. To test this hypothesis, we propose two specific aims. In Aim 1, we will assess the chemical transfor- mations of Der p 1 (the major HDM allergen), HDM, and genuine house dust via chromatography-mass spec- trometry, and associated changes to each mixture’s oxidative potential via an acellular assay. We hypothesize that diesel fuel components will sorb to HDM and, when the mixture is exposed to ozone, it will undergo oxida- tion, altering both HDM-sorbed diesel hydrocarbons and HDM’s protein composition, thus increasing the mix- ture's oxidative potential. In Aim 2, we will evaluate the impact of chemically contaminated HDM on a mouse model of AAI. We hypothesize that HDM chemically transformed by diesel fuel components and ozone will worsen AAI and be associated with increased oxidative stress markers. Our approach includes comparing standard AAI measures, such as bronchoalveolar lavage cell counts, cytokine expression, and airway hyperre- sponsiveness in mice exposed to chemically-treated HDM versus HDM alone. We will define and validate key cell types and pathways involved by performing single-cell RNA-seq on lung tissues post-exposure. This study advances our understanding of indoor transformation chemistry of dust-sorbed chemicals and HDM, address- ing the health impacts of these chemically processed species at cellular and systemic levels.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Breastmilk evolved over millions of years and has a fine-tuned composition that optimizes neonatal growth, development, and survival in diverse mammalian species. Breastmilk alternatives, such as infant formulas, are empowering, safe, and effective nutrient source for offspring. However, exclusive formula feeding is associated with an increased risk of developing many different metabolic, immunologic, and neurologic diseases in the offspring later in life, possibly due to missing components such as extracellular vesicles (EVs) that are normally found in breastmilk. Therefore, identifying novel components in breastmilk will increase our understanding of how breastmilk contributes to neonatal and infant health and may lead to strategies to improve the composition of formula. In my new preliminary studies, I found that murine breastmilk contains large quantalities of cell-free mitochondria that are predominantly derived from CX3CR1-postive macrophages in the lactating mammary gland. These cell-free mitochondria can be transported into some of the neonatal cells in the intestines during nursing. Moreover, prolonged breastfeeding or administering milk to Ndufs4–/– mice, a mouse model develops a mitochondrial disease called Leigh Syndrome (LS) after weaning, markedly reduced their disease severity and improved their survival. These findings provoke my central hypothesis that maternal cell-free mitochondria are released into breastmilk to deliver the full complement of mitochondria- associated nutrients and to support mitochondria metabolism in the offspring. I will test this hypothesis in two aims. In Aim 1, I will determine how cell-free mitochondria contribute to the nutrient composition of breastmilk. In Aim 2, I will establish how cell-free mitochondria in breastmilk contribute to normal development and limit the progression of LS in the offspring. This proposal is highly innovative because it will (1) identify cell- free mitochondria as a previously unknown breastmilk component that shapes the milk’s nutrient composition, (2) define the function of breastfeeding-associated mitochondria transfer in supporting neonatal metabolism, and (3) establish extended breastfeeding or mitochondria-supplemented formula as novel therapies for inherited mitochondrial diseases. These studies will increase our understanding of lactation physiology, the mechanisms of how breastmilk contribute to the growth and development of offspring, and provide novel insights on improving the composition of infant formula. I will complete the mentored phase (K99) of this project by leveraging the intellectual and scientific resources of Dr. Jonathan R. Brestoff’s Lab (one of the world’s leading labs studying mitochondria transfer and cell-free mitochondria), with co-mentorship by Dr. David Pagliarini (a prominent mitochondrial biologist) and an exceptional set of scientific and career advisors and collaborators. This project will allow me to transition to independence (R00 phase) to launch my career at a leading academic medical center in the United States.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT Bacterial vaginosis (BV) is a common, recurrent vaginal dysbiosis affecting nearly 30% of reproductive-aged women in the U.S. It involves a loss of protective Lactobacillus and an overgrowth of anaerobes such as Gardnerella and Prevotella, leading to elevated levels of BV-associated toxins with major implications for gynecological health and fertility. Notably, 37.4% of women with unexplained infertility are diagnosed with BV, yet the mechanisms remain poorly understood. The female reproductive tract fluid is essential for triggering sperm capacitation, a key maturation step for fertilization. However, the impact of BV on this process has been largely overlooked. My recent findings (Bhagwat et al., 2025, in press) reveal a novel mechanism by which BV toxins impair sperm function. Specifically, lipopolysaccharide (LPS, a toxin made by Prevotella) and vaginolysin (VLY, a Gardnerella toxin) interfere with sperm capacitation and reduce sperm fertility in vitro. Both toxins impair sperm hyperactivated motility, and acrosomal exocytosis (AE) in mouse and human sperm, and trigger premature and irreversible Ca2+ influx, which may result in sperm damage. This proposal aims to define the molecular mechanisms of BV-induced sperm dysfunction and assess the in vivo consequences of BV toxins on fertility, aligning with NICHD’s 2025 mission to improve reproductive health. I hypothesize that BV toxins impair sperm capacitation by dysregulating Ca2+ signalling, leading to impaired sperm hyperactivation and AE, disrupting sperm migration in the female tract and thereby impairing sperm fertilizing ability. During the K99 phase, I will: (1) Elucidate the mechanisms by which LPS and VLY induce Ca2+ influx. Since my preliminary data indicate that LPS-mediated Ca2+ influx requires both the TLR4 receptor and the TRPV4 Ca2+ channel, I will determine the contribution of these signalling receptors to the LPS response, by measuring single-cell Ca2+ dynamics, cAMP, and phosphorylation of PKA-PI3K-GSK3α pathway proteins in sperm from TLR4 and TRPV4 knockout mice. (2) Elucidate the mechanism of Ca2+ entry by VLY using scanning electron microscopy to examine VLY-induced pore formation and test the roles of membrane cholesterol and CD59 in this process. (3) Assess in vivo sperm function in a mouse BV model and wild-type mouse sperm exposed to physiologically relevant levels of LPS and VLY, by quantifying sperm migration, motility, AE, and fertility in the female reproductive tract. (4) Quantify LPS and VLY in vaginal specimens from women with and without BV and test their effects on human sperm motility, hyperactivation, and AE. During the R00 phase, I will model BV-associated infertility using a human vagina- and cervix-chip system. These chips will be colonized with Gardnerella and Prevotella, and incubated with sperm to evaluate motility, hyperactivation, AE, and mucus penetration under BV-mimicking conditions. This research will define how BV impairs sperm function, provide mechanistic insight into BV-associated infertility, and establish a high-throughput platform for evaluating human sperm responses to dysbiotic reproductive tract conditions.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Pelvic organ prolapse (POP) is an abnormal descent of the female pelvic organs that protrudes to, or through, the vaginal canal. Prolapse is caused by a loss of structural integrity of the pelvic organs and their connective tissue support and coincides with a reduction in vaginal elastic fibers either through enzymatic (as with age) or mechanical (as with vaginal delivery) disruption and fragmentation. One-in-eight women (13%) will require a surgical intervention to restore normal pelvic anatomy during their lifetime and over one-third of these procedures (38%) will fail or induce complications due to either insufficient native tissue strength or biomechanical mismatches in graft and adjacent tissue properties. The development of new and effective therapies for improving prolapse surgery outcomes is slow and the FDA mandate to stop selling and distributing transvaginal meshes underscores the need to study and develop innovative solutions. Currently, prolapse is treated surgically and there are no viable pharmaceutical treatment options focused on targeting the underlying cellular and molecular consequences of POP onset and progression in connective tissues. Towards this end, we propose a first-of-its-kind biologically motivated treatment that seeks to facilitate de novo vaginal elastogenesis. Motivated by prior findings of its use in the cardiovascular system, the goal of this work is to optimize the local administration of minoxidil, a promising ATP-dependent potassium channel opener, to the vaginal wall as a biologic therapeutic to improve pelvic floor function and integrity in mice. Our preliminary data shows that minoxidil treatment increased tropoelastin gene expression by primary murine vaginal cells and upregulated elastin protein secretion (soluble in media) and deposition (insoluble in tissue) in cultured vaginal explants. Finally, daily in vivo administration of minoxidil via local injections in the posterior vaginal wall led to an ~50% increase in elastic fiber area in the wall after seven days. To date, we have only explored a single concentration and delivery method for vaginal minoxidil treatment. Thus, we hypothesize that systematic investigation of dosage, delivery method, and treatment duration has the potential to optimize the in vivo elastogenic effects of vaginal minoxidil treatment. In this proposal, we will use biomechanical and biochemical assays to investigate the ability of increasing concentrations of local minoxidil injections to alter vaginal elastin production in female CD-1 mice of varying ages and parity status (Aim 1) and explore the efficiency of three separate routes of in vivo minoxidil delivery (local injection, local thermosensitive hydrogel, and systemic oral) on vaginal elastic fiber synthesis (Aim 2). Finally, we will explore the potential for minoxidil to promote elastic fiber persistence in the vaginal wall after treatment cessation (Aim 3). Outcomes from this work will demonstrate the therapeutic potential for minoxidil to generate vaginal elastic fibers in vivo, thereby improving biomechanical stability of the vaginal wall. We expect these results to have an important positive impact on women’s health and the management of pelvic floor disorders by yielding the first non-invasive biologic treatment to prevent or halt the progression of prolapse.

NIH Research Projects · FY 2026 · 2026-06

This proposal aims to undertake a next-generation cortical cartography effort by generating a more accurate multi-modal atlas of human cortical areas, studying the individual variability of these cortical areas and their evolutionary lineages, validating and refining non-invasive methods of cortical connectivity, and enhancing our widely used software tools specially designed for cortical analysis. Much new data is now available for improving our highly impactful multi-modal atlas of human cortical areas published in 2016, including higher resolution and different fMRI scans, higher resolution and better architectural measures, and non-MRI-based measures like spatial transcriptomics which assess both gene expression patterns and cell type distributions in cerebral cortex. Additionally, individual variability and subareal organization of these cortical areas have not been well charac- terized; however, understanding these aspects is critical for the new atlas to have maximal clinical impact, as demonstrated by our recent highly successful clinical-translational applications of the current atlas. Another ma- jor objective is to better understand the evolution of human cortical areas relative to intensively studied macaque and marmoset monkeys by identifying areas common to all 3 species and areas found only in the human lineage. This will enable better translation of Non-Human Primate (NHP) invasive experiments to humans. NHPs also enable quantitative neuroanatomical measures of brain connectivity, providing an invaluable gold standard for validating noninvasive MRI-based methods for estimating cortical connectivity that are widely used in humans. Identifying the best-performing methods in NHPs will accelerate progress in developing more accurate non- invasive connectivity methods for humans. Finally, our widely used tools for multi-modal cortical visualization (Connectome Workbench), data sharing (BALSA neuroimaging study results repository), and analysis (the CIFTI file format and standard space that enables precise combined cortical and subcortical analysis) must both be maintained and extended in multiple ways to enable the new datatypes to be best combined with existing data. Altogether, this project has five key deliverables. (1) A version 2.0 of our atlas of human cortical areas, including elucidation of their subareal organization. (2) Characterizing the individual variability of human cortical areas, including their size, shape, and position and whether all humans have the same set of areas or whether some are missing areas and others have extra areas. (3) Identifying which cortical areas have homologues in NHPs and which are human specific. (4) A critical evaluation in macaques of the non-invasive measures of functional and structural connectivity that are widely used in humans to determine which methods perform best at replicat- ing invasive connectivity in the macaque and then producing human cortical connectomes using the best meth- ods. (5) Improvements to our widely used tools for cortical visualization, data sharing, and analysis. Completing these aims will accelerate progress towards understanding the cerebral cortex and enable more rapid translation of basic science insights and neuroanatomical maps to clinical practice for diseases involving the cerebral cortex.

NIH Research Projects · FY 2026 · 2026-06

Project Abstract: Mitchell Syndrome is a progressive childhood-onset neurodegenerative disorder characterized by sensory ataxia, hearing loss, skin changes, and eventual paralysis and encephalopathy, typically leading to death within the second decade of life. The first patient with Mitchell Syndrome was treated at Washington University in St. Louis (WashU), which makes WashU uniquely situated for researching this rare disease. WashU sees around 30% of known patients, has characterized the disease's natural history, and has extensive resources such as post-mortem tissues and a biofluid biobank. Our long-term goal is to develop effective treatments for Mitchell Syndrome. The disease is caused by an autosomal dominant variant in ACOX1, leading to a gain-of-function in the acyl-CoA oxidase 1 (ACOX1) protein. Our preliminary studies indicate that the variant impacts both sensory neurons and oligodendrocytes. Patient-derived induced pluripotent stem cells (iPSCs) offer a scalable, homogeneous platform for modeling rare diseases and evaluating precision-medicine therapeutics. We aim to develop iPSC-derived models of oligodendrocytes and sensory neurons to recapitulate Mitchell Syndrome as a tool to evaluate potential treatments. Preliminary data from patient-derived iPSC lines show transcriptional and metabolic abnormalities linked to the disease variant. We propose two specific aims: Aim 1: Evaluate ACOX1 gain-of-function in iPSC-derived oligodendrocytes. Aim 2: Investigate ACOX1 gain-of-function in iPSC-derived sensory neurons. Mitchell Syndrome intertwines lipid metabolism, oxidative stress, and neuronal/glial degeneration. This project aims to provide essential models for therapeutic evaluation, leveraging WashU's unique expertise and resources. At the culmination of this project we will have two scalable, disease-relevant, human models of Mitchell Syndrome that can be used for mechanistic studies, therapeutic development, and biomarker exploration – critical steps on the way to treat this lethal and tragic disease.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Mitochondria are multifaceted organelles responsible for cellular energy production and nutrient catabolism, efficiently performing these tasks as distinct metabolic compartments. Such compartmentalization requires fine- tuned regulation of not only catabolic enzymes but also their associated co-factors. One such compartmentalized co-factor is nucleotide adenosine diphosphate phosphate (NAPDH), which enables numerous metabolic pathways within mitochondria including unsaturated fatty acid oxidation, lysine catabolism, type II fatty acid synthesis, and proline biogenesis, amongst others. Due to its broad utilization, perturbation of NADPH synthesis would have widespread, detrimental effects on mitochondrial metabolism. Indeed, human mutations in a key NADPH-producing enzyme, NADK2, can lead to Progressive encephalopathy with leukodystrophy due to DECR deficiency, an ultra-rare disease presenting with failure to thrive, pronounced hypotonia, and microcephaly. However, a recently identified patient harboring hypomorphic NADK2 alleles presented with a much milder disease profile and only select metabolic defects, suggesting that full loss of NADK2 broadly compromises mitochondrial metabolism, while partial loss of NADK2 affects only select pathways. We hypothesize that mild- to-intermediate loss of NADPH production disproportionately affects select pathways to spare more “essential” pathways, leading to a “priority code” of mitochondrial co-factor usage. As NADK2 represents a central NADPH- producer within mitochondria, we propose that an in-depth functional assessment of disease-associated NADK2 variants will illuminate a “priority code” for mitochondrial NADPH allocation, while simultaneously providing a mechanistic framework to understand disease heterogeneity. In this proposal, we will generate a series of cell lines that harbor NADK2 variants of uncertain significance with different predicted mutational burdens: low, moderate, or high. We will first establish and characterize these mutants by expressing each variant in NADK2 knockout HAP1 cells and measuring the extent to which these mutants can rescue mitochondrial respiration as well as organellar and cellular NADPH levels. We will then profile these cell lines with multiomic analyses of lipids and metabolites to understand the extent to which metabolic pathways become compromised upon limiting NADK2-produced mitochondrial NADPH. Completion of the proposed work will, as a proof of principle, demonstrate the power of cellular models of rare disease, as well as the breadth of dysfunction associated with specific mutational burdens, which could inform accurate diagnoses or therapeutic options for patients with NADK2 or DECR deficiency. Furthermore, these cells will provide a comprehensive toolkit to study how NADPH perturbations affect mitochondrial metabolism, providing a rich dataset of the dynamic nature of organelle-wide NADPH-consuming pathways.

NIH Research Projects · FY 2026 · 2026-06

Project Summary The goal of this project is to understand the mechanisms and potential therapeutic targets of a novel inborn error of immunity (IEI) caused by variants in toll like receptor 8 (TLR8). We recently identified genetic variants in TLR8 leading to gain-of-function (GOF) of the encoded protein as the cause of a new IEI presenting with profound neutropenia with recurrent infections, lymphoproliferation, T and B cell abnormalities, and bone marrow failure. Toll-like receptor 8 (TLR8) is an endosomal TLR encoded on the X chromosome that recognizes single-stranded RNA (ssRNA) and is expressed in neutrophils and other myeloid cells, including myeloid progenitors. Relatively little is known about human TLR8, largely because the murine equivalent differs in structure and ability to sense ssRNA. There is no available targeted therapy for patients with TLR8 GOF; however, patients have benefited from hematopoietic stem cell therapy, confirming that disease is driven by the immune system. There is a gap in our understanding of how GOF in TLR8 protein function leads to the clinical features of this newly recognized disease. As TLR8 expression is myeloid-specific and, in most cases of TLR8 GOF disease, expressed in a mosaic fashion (with approximately 10-20% of cells harboring TLR8 variants), we hypothesize that myeloid cells expressing mutant TLR8 produce inflammatory cytokines leading to T cell activation, and this inflammatory state and dysfunctional T cells contribute to the disease in patients including lymphoproliferation and bone marrow failure. Given the differences between mouse and human TLR8, we recently generated transgenic mice that conditionally express either WT or GOF human TLR8. Herein, we will use these novel models to address several gaps in our knowledge of TLR8 biology and TLR8 GOF disease, as well as to test potential therapies. In Aim 1, we will identify mechanisms of TLR8 GOF disease, using our transgenic mouse models and human cell xenografts to determine how TLR8 GOF myeloid cells influence hematopoiesis and T cell development and identify putative disease-driving cytokines. In addition, we will formally test the role of T cells in the disease pathogenesis by crossing TLR8 GOF mice to T-cell deficient mice and determining the effects on hematopoiesis, cytokine production and disease progression. In Aim 2, we will identify therapeutic targets of TLR8 GOF disease using inhibitors of TLR8 signaling and blocking candidate cytokines in our transgenic mice. Together, these studies will lead to an increased understanding of the pathology of TLR8 GOF and will identify therapeutic targets. The long-term goal of this application and our work is to develop a mechanistic understanding of how TLR8 GOF alters the immune response to enable improved therapies for patients.

NIH Research Projects · FY 2026 · 2026-06

Identification and Characterization of Mutation-Induced Alternative Splicing Events in Cancer Using Multi-Omics Data Project Summary The goal of this project is to discover mutation-induced alternative splicing events (MAS), understand their functional relevance, and identify neoantigens arising from these events to advance cancer immunotherapy. Large-scale sequencing efforts such as The Cancer Genome Atlas (TCGA) have primarily focused on identifying driver mutations in tumors, including single amino acid changes, insertions, deletions, and alterations that truncate or elongate wild-type protein sequences. However, traditional DNA mutation annotations often rely on canonical transcripts and may overlook alternative splicing events, and some mutations, such as synonymous changes, are considered silent despite their potential impact on splicing. Previously, we developed the MiSplice pipeline to detect mutation-induced splice sites and, when applied to TCGA data, identified thousands of somatic mutations that create cryptic splice sites. In this proposal, we aim to systematically investigate mutation-induced alternative splicing and its functional relevance in cancer, while also identifying the resulting neoantigens by leveraging data from the Clinical Proteomic Tumor Analysis Consortium 3 (CPTAC-3), which includes comprehensive exome sequencing, RNA-Seq, and mass spectrometry data for 774 tumors across seven cancer types, as well as 126 prospective breast cancer samples from CPTAC-2. We hypothesize that mutation-induced alternative splicing plays a significant role in cancer etiology and that the associated neoantigens can serve as novel immunogenic peptide candidates for cancer immunotherapy. We propose to test these hypotheses through two specific aims. Aim 1: Identify mutation-induced alternative splicing events and assess their functional relevance in cancer using multi-omics data (Years 1 & 2). We will use MiSplice to detect mutation-induced alternative splicing events from CPTAC data and evaluate their impact by analyzing changes in protein and phosphorylation expression, as well as pathway activation. Aim 2: Identify neoantigens arising from mutation-induced alternative splicing events with mass spectrometry support (Years 1 & 2). We will construct a tailored protein database that integrates both reference proteins and mutant proteins generated from MAS events. Using state-in-art tools such as PepQuery, we will search for corresponding peptides in the mass spectrometry data, enabling identification of mutant proteins. Neoantigen peptides derived from these expressed mutant proteins will then be prioritized as candidates for cancer immunotherapy.

NSF Awards · FY 2026 · 2026-06

From June 14–20, 2026, Washington University in St. Louis will host the 48th Summer Symposium in Real Analysis (SSRA48), an official satellite conference of the International Congress of Mathematicians. For nearly five decades, this annual global symposium has brought together mathematicians from around the world to share ideas, train young researchers, and foster long-term collaborations. By returning the Symposium to the United States for the first time since 2019, SSRA48 will significantly expand access for graduate students and early-career researchers, particularly those in North America. The conference will provide these participants with direct exposure to world leaders in real analysis, a core area of mathematics whose ideas underpin modern developments in the physical sciences, engineering, and data science. The International Congress of Mathematicians satellite designation highlights the meeting's global significance and visibility.  SSRA48 will focus on recent advances in real analysis and closely related fields, including harmonic analysis, geometric measure theory, partial differential equations, and analysis on metric spaces. The plenary speakers are leading experts in real analysis, and their work includes connections to algorithmic information theory, learning theory, signal processing, time-frequency analysis, dynamical systems, and fractal geometry. In addition to the plenary lectures, the program will feature contributed talks by researchers at all career stages, a poster session emphasizing graduate student participation, and a problem session designed to stimulate new collaborations. By emphasizing early-career participation alongside established leaders, SSRA48 will advance cutting-edge research while supporting the professional development of the next generation of mathematicians. The conference website is https://www.math.wustl.edu/ssra48/ This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2026-05

Project Summary Invasive fungal sinusitis (IFS) is a devastating form of fungal infection with high morbidity and mortality that affects immunocompromised patients. There is a critical need for improved treatment options in this highly vulnerable population. Very little is currently understood regarding the pathophysiology of IFS; the mechanisms mediating fungal invasion, specifically those related to fungal virulence and mucosal barrier degradation, remain unclear. Prior studies have reported that Aspergillus, a common fungal species seen in IFS, can persist and invade in pulmonary disease due to mutations that enable survival in hypoxic environments. Our preliminary studies evaluating sinonasal tissue found that matrix metalloproteinases (MMPs), specific proteases that are secreted in response to infection, are upregulated in IFS patient tissue as compared to tissue from immunosuppressed patients without IFS. Levels of tissue inhibitors of MMPs (TIMPs) were found to be proportionally lower in IFS tissue, leading to abnormal MMP/TIMP ratios. Loss of MMP inhibition can lead to aberrant extracellular matrix degradation. Interestingly, published data has shown that non-invasive fungal sinusitis tissue exhibits alterations in markers of epithelial barrier integrity, as compared to healthy controls. Together, these data prompted us to investigate the contributions of fungal virulence, protease secretion, and tissue permeability and dysregulation to IFS. In Aim 1 of the proposed prospective study, we will use RNA sequencing to evaluate transcriptomic differences and an in vitro epithelial permeability model to evaluate and compare phenotypic differences in cultured fungal isolates and fungal supernatant between patients with histopathologically confirmed IFS (Cohort 1) and patients with non-invasive fungal sinusitis (Cohort 2). In Aim 2, we will compare tissue samples between both of these cohorts using single cell RNA sequencing, immunostaining, and western blotting techniques to investigate whether IFS tissue contains decreased lymphocytes, increased macrophages responsible for secreting MMPs, and altered epithelial integrity markers leading to decreased compensatory TIMP secretion. The long-term goal of this research effort is to understand the mechanisms that mediate IFS spread in immunocompromised patients to support the development of targeted therapies. The proposed research study is directly linked to my proposed training activities for this K23 Mentored Patient-Oriented Research Career Development Award. My overall career goal is to transition to an independent research career as a surgeon-scientist and to become a recognized expert in the field of fungal sinusitis. I have assembled an interdisciplinary mentorship team with expert investigators tailored to both the proposed research and my training and mentorship needs. Together, we have developed a gap-based training plan that combines formal didactic training, mentored training activities, and participation in a range of meetings and conferences to achieve my training objectives of gaining essential expertise in (1) mycology, and (2) immunology, particularly in relation to sinonasal mucosa, to support my transition to independence.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY For successful fertilization in vivo, mammalian sperm must first undergo capacitation, a maturation step that enables hyperactive motility and acrosomal exocytosis. Hyperactive motility is triggered by calcium influx through the sperm-specific calcium channel CatSper, but premature CatSper activation can lead to calcium overload, channel degradation, and loss of fertilizing capacity. To prevent this, CatSper must remain inactive until sperm reach the oviduct. Prevention of premature CatSper activation in the male reproductive tract has long been attributed to acidic epididymal pH, which inhibits CatSper. However, the factors that prevent CatSper activation before sperm reach epididymis and after they leave it remain unknown. Based on our recently published study in a murine model, this project will test the central hypothesis that human CatSper activation is regulated within the male and female reproductive tracts by four key factors: pH, temperature, polyamines, and progesterone. In this model, the low temperature of the testis and the acidic pH of the epididymis maintain CatSper in a closed state. When sperm mix with seminal plasma, they encounter polyamines, which further inhibit CatSper activity, even as the sperm travel through the warm and slightly alkaline environment of the cervix and oviduct. During sperm capacitation in the oviduct, and after prolonged exposure to oviductal fluids, polyamines dissociate from sperm. At this stage, CatSper is activated by high temperature, alkaline pH, and factors released by the ovulated egg, such as progesterone. This hypothesis will be tested in three specific aims. Aim 1 is to define the mechanism by which CatSper is regulated by temperature in human sperm. This aim will test the hypothesis that temperature alters human CatSper activity by influencing its voltage gating, pH- and progesterone- sensitivity. Aim 2 is to determine the effect of these four factors on CatSper impact on sperm motility and intracellular calcium concentrations in both murine and human sperm. These two aims test the hypothesis that the seminal plasma components inactivate CatSper and greatly reduce its temperature sensitivity, thereby protecting CatSper from activation until sperm reach the oviduct. Aim 3 is to determine the molecular mechanisms underlying infertility in patients with varicocele. This aim will test the hypothesis that, in patients with varicocele, high testicular temperature, lack of acidic pH, and absence of protective polyamines mimic conditions that sperm normally only encounter in the oviduct. Thus, sperm CatSper is prematurely activated in the testis, leading to calcium overload and shortened sperm life span. Completion of these aims will rely on the unique abilities of the investigators’ laboratories to conduct electrophysiological measurements, motility analysis, and calcium imaging in mouse and human sperm. The proposed work will reveal key physiological regulators of CatSper, and may reveal the molecular mechanism underlying infertility in patients with varicocele.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Glioblastoma (GBM) is a brain tumor that causes neurological deterioration and death in most patients within 2 years. The long-term goal of this project is to develop a new treatment approach for GBM, by expanding peripheral effector immune cells with interleukin (IL)-7, and recruiting them into the tumor microenvironment with an oncolytic virus. In mouse models of GBM, we have observed an increase in survival when we combine long- acting IL-7 with oncolytic virus. Our central hypothesis is that combination therapy (a) improves CD8+ T cell trafficking to the TME, expansion, and anti-tumor function; (b) decreases myeloid and T regulatory cell driven immunosuppression in the TME; and (c) is effective when paired with standard-of-care radiation. Collectively, we term this concept “expand and pull” (expanding the immune system, recruiting it to the TME, and activating an anti-tumor response), and we will pursue it in two Aims. First, we will determine how long-acting IL-7 plus oncolytic virus therapy improves CD8+ T cell brain trafficking and function, and how it alters the tumor microenvironment. Second, we will determine whether long-acting IL-7 plus oncolytic virus therapy enhances standard-of-care outcomes and generates tumor infiltrating lymphocytes (TILs) that can treat recurrence. If successful, Aim 1 will determine the mechanism(s) by which IL-7 plus oncolytic therapy is effective, (and establish ways to improve it), and Aim 2 will promote translation to a clinical trial.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract Nine NIH-funded investigators with similar but independent super-resolution microscopy needs are requesting funding to acquire an Abberior Mirava 3D STED microscope to enable imaging of molecular topographies in fixed or live tissues and cells at unprecedented resolution at the scale of tens of nanometers with fluorescent probes. This platform will be maintained in the Washington University Center for Cellular Imaging (WUCCI), an institution-wide shared technology resource. There is no instrument like this at WUCCI or anywhere else on campus. One huge advantage of this system is that it works just like a confocal microscope in terms of the field of view, imaging depth, and data acquisition interface. As such, many investigators will be ready to utilize it immediately. Sample preparation is nearly identical to conventional confocal microscopy, except for the use of specific fluorescent probes, and care taken to use mounting or immersion media and cover slips with matching refractive indexes to enable maximum resolution. STED technology breaks the traditional barrier of optical microscopy set by the diffraction-limited point spread function (PSF) by PSF engineering, achieving 40 nm resolution, exceeding the resolution of the already heavily subscribed Nikon NSPARC and Zeiss Airyscan. The major user group comprises nine investigators from 6 departments who will utilize this microscope to enable a wide range of basic and translational research studies. Scientific goals include understanding primary cilia, the role of actin cytoskeleton dynamics in intracellular networks, energy and lipid storage and lipid droplet or to resupply membranes with lipids, mechanobiological mechanisms underlying kidney glomerular filtration, glutamate receptor geography at cochlear synapses, receptor dynamics in pancreatic islet hormone secretion and neuroinflammation biomarkers for biological imaging and chemotherapeutics. Given its location in a busy core facility, we expect this microscope to be impactful to many research programs beyond the initial major user groups. The expertise and institutional support for this instrument are excellent. Dr. Mark Rutherford, Associate Professor of Otolaryngology, used STED microscopy extensively during the historical development of this technology. Dr. Rosa-Molinar and the WUCCI staff have a long-standing track record of training NIH- funded researchers to optimize their use of state-of-the-art microscopy methods. In support of this S10 grant application, the Washington University School of Medicine will commit $176,647 of matching funds for the acquisition of the instrumentation plus an additional $125,000 operational support ($25,000 per year for five years) to ensure the long-term success of this equipment.