Washington University

NIH Research Projects · FY 2026 · 2019-03

Project Summary The fundamental goal of the proposed Washington University/Siteman Cancer Center (WU/SCC) NCTN Lead Academic Participating Site UG1 is to 1) foster scientific leadership and mentorship in the NCTN, 2) participate in the development of NCTN therapeutic, correlative, and cancer control trials, and 3) maintain strong accrual to clinical trials across multiple disease sites, including those that are molecularly driven and those for rare cancers. WU/SCC investigators have a long history of enthusiastic participation in the former cooperative group program and continue to play key roles in the current NCTN program, specifically the Alliance for Clinical Trials in Oncology, ECOG-ACRIN Cancer Research Group, and NRG Oncology. WU/SCC will have 4 UG1 co- Principal Investigators comprising the senior leadership team including Nancy L. Bartlett, MD (Professor, Medical Oncology), Premal Thaker, MD (Professor, Gynecologic Oncology), Clifford Robinson, MD (Professor, Radiation Oncology) and Barry A. Siegel, MD (Professor, Radiology). The WU/SCC senior leadership team is composed of four legacy members with extensive cooperative group and NCTN experience (Bartlett, Thaker, Robinson and Siegel). We feel this consistency will bring continued success to the NCTN program. The entire leadership team is committed to providing guidance for an institution-wide NCTN UG1. WU/SCC has consistently demonstrated robust accrual to NCTN trials, accruing 1217 patients between March 2019 and August 2024. During this same interval, 764 patients had biospecimens collected and 32 patients were accrued to advanced imaging studies. The strength of the WU/SCC genomics, proteogenomics, and molecular imaging programs in combination with the large institutional biorepositories position us to make significant and novel contributions to the scientific mission of the NCTN. WU/SCC investigators remain highly invested in NCTN translational and clinical research in AML, lymphoma, lung cancer, breast cancer, sarcoma, uterine cancer, molecular imaging, and radiation oncology. In addition, our large referral base the expansion of Siteman Cancer Center to two additional satellite locations (North St. Louis County and Shiloh, IL) will enhance our ability to accrue patients to nearly all disease site protocols, including rare tumors. A new cancer-only 650,000 sq. ft. ambulatory care facility on the Washington University Medical Campus opened to patients in September 2024. The Gary C. Werths Building at Siteman Cancer Center represents a transformational opportunity to enhance multidisciplinary care at SCC, increase accrual on clinical trials, and improve patient experience through centrally and co-located clinics, treatment areas, pharmacy, and diagnostic services. This institution-wide UG1 will facilitate enhanced communication and collaboration among disease-site, translational, and modality specialists and improve our potential for contributions to the NCTN operation.

NIH Research Projects · FY 2026 · 2019-03

Project Summary Mass spectrometry (MS) based footprinting is emerging as a powerful means to answer biological questions about membrane proteins (MPs), which participate in almost all physiological processes and represent more than 60% of drug targets. This approach affords sufficient structural information for the dynamic, native conformations and interactions of MPs in cells, which are beyond the reach of traditional structural methods (e.g., cryo-EM and crystallography). This bottom-up MS footprinting is complementary to but potentially more informative than top-down native MS, which does not provide spatial resolution for MPs and is conducted in the nonnative gas phase. Here we propose to continue development of novel MS footprinting methods in live cells and native membranes. Our objective is to design, prepare, test, and improve chemical probes that provide high footprinting coverage. We will then apply them to reveal drug interactions and cellular trafficking regulation of a glucose transporter, GLUT1, a prominent anticancer drug target and a model MP representing ~ 25% of known transport proteins. MS footprinting of MPs, however, poses three major challenges. 1) MPs, which are hydrophobic and buried in lipid bilayers, are resistant to traditional probes (e.g., HDX, •OH radicals) that penetrate poorly and give insufficient labeling. 2) Aliphatic side chains of transmembrane regions contain C–H and C-C bonds that are unreactive with most chemical probes. 3) The footprinting needs to be conducted in cells or membranes to maintain native conformation and interaction of MPs. Our hypotheses are: (1) Complementary modifications of C-H and X–H bonds by free radicals produced photochemically and by nucleophilic reagents maximize footprinting coverage. (2) Tuning the hydrophobicity of the reagents or their precursors allows access to membrane-embedded MPs. (3) Novel membrane fusion techniques introduce inert footprinters into live cells and native membranes for subsequent photoactivated footprinting. Our hypotheses are built on extensive preliminary data. Three years of funding supported publication of 18 papers in high-profile journals. A significant example describes laser activation of TiO2 nanoparticles attached to liposomes to generate high local concentrations of radicals. Simultaneous membrane poration permits radical entry to footprint with sufficient structural resolution that reports the ligand-binding sites and rocker-switch motions of GLUT1. Building on these successes, we will pursue two specific aims: (1) develop new chemical probes for MS footprinting of MPs; and (2) conduct comprehensive footprinting in native membranes and live cells to reveal anticancer drug interactions and trafficking regulations of GLUT1. Our innovative footprinting coupled with bottom-up MS proteomics analysis will establish bio-orthogonal footprinters that afford comprehensive coverage of both hydrophobic and hydrophilic regions of MPs and reveal drug interactions and structural regulation of human MPs under inarguable native settings. The impact of the proposed approach should readily expand because MS-based footprinting can be broadly applied in structural proteomics to expedite drug discovery and structural studies of cellular processes.

NIH Research Projects · FY 2026 · 2019-03

ABSTRACT Short gut syndrome (SGS) results from surgery that removes a significant length of small intestine in response to different conditions, like necrotizing enterocolitis. In children, the mortality associated with SGS is ~25%, making it one of the most lethal conditions in infancy and childhood. After surgery, remodeling of the remaining bowel segment can facilitate absorptive function and survival. Nonetheless, some patients require long-term parenteral nutrition. Beyond loss of absorptive capacity, a significant complication in SGS patients that survive bowel resection is intestinal failure-associated liver disease (IFALD), regardless of whether patients receive nutrition enterally or parenterally. Clinically, therapies designed to target IFALD after SGS remain elusive. In the first round of funding of this continuation application, The Warner and Randolph lab have shown in mouse models of small bowel resection that gut-derived LPS transiting to the liver from the portal vein initiatesTLR4 signaling in liver macrophages that ultimately drives onset of fibrosis. A significant break on LPS-TLR4 interaction occurred by the binding and neutralization of gut-derived LPS to portal vein high density lipoprotein (HDL). HDL is synthesized in the liver and is also generated by enterocytes in the mice or human ileum, where it depends on expression of the polypeptide apoA1 and the lipid transporter ABCA1 that donates the first lipids to the nascent HDL particle. Loss of enterocyte ABCA1 greatly reduced portal vein HDL, allowing for enhanced LPS activity to reach the liver and exacerbate liver injury after small bowel resection. Conversely, when we increased intestinal HDL by giving the liver X receptor (LXR) agonist GW9365 orally at a low dose to trigger HDL production in the intestine but avoid direct LXR agonism of the liver, the drug was effective in reducing the onset of liver fibrosis in a manner that was dependent upon enterocyte-derived ABCA1. These findings raise the concept that intestinal epithelial activation of the transcription factor LXR may provide therapeutic benefit toward treating IFALD. This grant is designed to consider this concept with the intent to further define the role of LXR and HDL in protecting the liver in SGS models and to move in a translational direction that may support later clinical development of a therapeutic to treat IFALD. We will study how LXR agonism protects the liver in a mouse model of SGS using a novel orally restricted LXR agonist compound WUSTL0717 synthesized by Principal Investigator Elgendy. Pharmacokinetic analysis in mice reveals that WUSTL0717 is indeed restricted to the intestine. To further consider the translational potential of WUSTL0717, Principal Investigator Ajay Jain will provide expertise in SGS studied in piglets. One major advantage of incorporating a piglet model into studies of SGS is that piglets can be provided parenteral nutrition like humans and thus model aspects of the disease not easily studied in mice. Dr. Jain has shown that parenteral nutrition in piglets leads to bacterial overgrowth in the small bowel, resulting in increased LPS translocation to the liver and thereby recapitulating the scenario and possible mechanisms described above in mice.

NIH Research Projects · FY 2025 · 2019-02

Summary Atherosclerosis is a progressive disease characterized by the development of lipid-rich, inflammatory plaque lesions within vessel walls. It is the underlying basis of cardiovascular diseases including myocardial infarction, stroke, and peripheral arterial disease. However, the ability to reliably detect the vulnerable plaque and identify high-risk patients has been a challenge. Further, there is no imaging agent to detect the eroded plaques, a less- known subtype accounting for one third of clinical events. Chemokines and chemokine receptors play important roles in atherosclerosis from initialization to clinical event by directing leukocyte trafficking. We have developed chemokine receptor targeted positron emission tomography (PET) imaging agents and demonstrated the specific detection of monocyte trafficking in vivo and track plaque progression and regression. To further explore the potential of these imaging agents for translation, we would like to propose a research program to develop novel PET tracers with potential to identify vulnerable plaques, detect plaque erosion, and more importantly to track the treatment response to improve patient outcome. Specifically, we will firstly optimize the design and synthesis of a portfolio of PET tracers targeting plaque-relevant targets including CCR2, CCR5, CXCR3, CD44, and TLR2 to improve the radiolabeling and scale-up capability through controlled radiochemistry and bioconjugate chemistry, and binding affinities by varying the charge, surface chemistry and polymer coating materials. Secondly, we will perform in vivo biodistribution studies and PET imaging in atherosclerosis progression/regression and complication mouse models, as well as rabbit atherosclerosis models to assess the imaging specificity, sensitivity, and capability to track the immune cells in vivo and correlation with targets expression and plaque characteristics. Thirdly, we will assess the capability of developed imaging probes to determine treatment response for improved outcome and binding to ex vivo human plaque tissue for future translation. We propose to submit multiple exploratory investigational new drug application to FDA and have two PET tracers ready for human trials at the end of grant period. The establishment of this research program will not only promote the development of targeted PET tracers for atherosclerosis translational imaging, but also broader applications in other diseases within the NHLBI mission.

NIH Research Projects · FY 2024 · 2019-02

Project Summary/Abstract Centrioles are conserved cellular organelles that template cilia for mediating cell signaling and motility, and recruit pericentriolar material to nucleate microtubules as part of the centrosome. Centrioles are extremely stable structures that persist over multiple cell cycles, and centriole number is controlled to ensure that each daughter cell inherits exactly two centrioles from its mother. Defects in centrosome and cilium function have been linked to a wide range of diseases, including cancer, microcephaly, and a set of syndromes known as ciliopathies. Within a centrosome, the older centriole in the pair templates the cilium, and together they form the centriole-cilium complex. Key features of this complex are the compound microtubules, which are a unique arrangement of microtubules linked together: three linked microtubules form the centriolar triplets, and two form the ciliary doublets. These compound microtubules occur only in the centriole-cilium complex, and are required for the structural integrity of the centriole and for protein trafficking in the cilium. Though the morphology of compound microtubules has long been appreciated, little is known about the mechanisms by which these microtubules form specifically at the centriole-cilium complex. In my postdoctoral work, I found that two members of the tubulin superfamily, delta-tubulin and epsilon-tubulin, are required for the centriolar triplet microtubules. The means by which these proteins act are unknown. Here, I propose to determine the mechanisms of compound microtubule formation through an integrated set of aims. Spanning both the mentored and independent phases, these aims will allow me to test the relative contributions of microtubule protofilaments themselves, as well as other proteins including delta- tubulin and epsilon-tubulin, to compound microtubule formation and stability. With the help of an outstanding collaborator and mentor team, I will train in research techniques, as well as skills for my career development. Together, this will create a strong foundation for an independent research career in understanding the roles of microtubule structures in human development and health.

NIH Research Projects · FY 2024 · 2019-01

PROJECT SUMMARY/ABSTRACT Development of effective therapies is an urgent, unmet medical need for patients with pancreatic ductal adenocarcinoma (PDAC). The advent of immune checkpoint antagonists such as anti-PD-1 and anti-CTLA4 antibodies has revolutionized treatment of some cancers but remains unsuccessful in PDAC. We and others showed that the tumor microenvironment (TME) of PDAC is rife with myeloid-derived suppressor cells including inflammatory monocytes (IMs) and tumor-associated macrophages (TAMs) that stifle the effect of chemotherapy and anti-tumor immunity. Excessive production of CCL2 in PDAC has shown to result in tumor growth, dissemination, local immunosuppression, and resistance to chemotherapy. Targeting a key chemotactic mechanism, the C-C motif chemokine ligand 2 (CCL2)/ C-C chemokine receptor type 2 (CCR2) axis, that draws these cells to the TME potentiates the efficacy of chemotherapy in preclinical mouse model and a clinical trial conducted at our institution, setting the premise to further confirm and optimize CCR2-targeted strategies in PDAC. We are in the process of opening a phase I/II clinical trial combining a CCR2/5 inhibitor, chemotherapy and anti-PD-1 agent. Realizing that not all patients will benefit from this regimen, a diagnostic tool capable of assessing CCR2 abundance while predicting and monitoring treatment response will be invaluable. CCR2 inhibitor slows tumor progression, prevents metastasis in mouse models of PDAC, and potentiates effect in patients with border-line resectable or locally-advanced PDAC (NCT01413022). We have developed a CCR2- PET tracer (64Cu-DOTA-ECL1i) and shown its sensitivity and specificity in imaging CCR2 in multiple preclinical inflammatory disease models and PDAC models and PDAC human specimens. Our PDAC PET imaging in genetic mouse model demonstrated early, sensitive, and specific detection of CCR2 in tumors. The first- in-man imaging showed low accumulation of 64Cu-DOTA-ECL1i in normal pancreas and liver (a common site of metastatic disease where CCR2-bearing IMs and TAMs infiltrate the pre-metastatic sites prior to establishment of metastatic clones) with rapid blood and renal clearance, indicating the potential of this PET tracer for CCR2 detection in PDAC patients. We hypothesize that 64Cu-DOTA-ECL1i can sensitively and specifically detect CCR2 in PDAC, track the variation following CCR2-targeted treatment, and likely prescreen PDAC patients for CCR2-targeted therapy. We propose to evaluate whether tumor uptake of 64Cu-DOTA-ECL1i correlates with tumor expression of CCR2 and response to standard chemotherapy in PDAC patients. We also will evaluate whether tumor uptake of 64Cu-DOTA-ECL1i predicts response to CCR2-directed therapy in PDAC patients treated with CCR2/5 inhibitor and chemo-immunotherapy. The successful completion of this grant will facilitate innovative means for clinical data interpretation, patient stratification, and therapy guidance.

NIH Research Projects · FY 2026 · 2018-12

ABSTRACT mRNA-based vaccines use lipid nanoparticles to transport mRNA-encoding antigens to the host cells. We have recently shown that SARS-CoV-2 mRNA vaccination in humans induces a robust plasmablast response in blood and a persistent GC reaction in the draining lymph nodes. This corresponded with enhanced anti-spike antibody avidity in blood and enhanced affinity and neutralization capacity of bone marrow plasma cell (BMPC)-derived monoclonal antibodies (mAbs). mRNA-based influenza vaccines could be a promising alternative to conventional influenza vaccine platforms because of their high immunogenicity. The following questions, however, are yet to be addressed: (1) does an mRNA-based influenza vaccination induce a robust and persistent GC reaction in humans? (2) if yes, what drives such persistence, and how different is that from the GC response induced by conventional influenza vaccines? (3) can an mRNA-based influenza vaccination induce a robust GC response in the elderly (>65 years old)? (4) does robust GC response to influenza vaccination in humans correlate with a more durable antibody response? (5) does an mRNA-based influenza vaccination induce a more sustained increase in the frequency of long-lived BMPCs compared to conventional vaccines? (6) can a vaccine-induced robust GC response overcome influenza antigenic imprinting in humans? (7) what are the molecular determinants that dictate the persistence of BMPCs? Addressing these questions will allow us to discern the cellular and molecular determinants associated with durable antibody responses to vaccination in humans. We will directly examine antigen-specific GC and long-lived BMPC responses induced after mRNA-based and conventional influenza vaccination in humans in the proposed studies. These responses will be examined in 18- 50 year-old and 65-80 year-old human adults. We will assess the persistence of the vaccine antigen in draining lymph nodes as a potential mechanism for driving GC longevity in humans. We will examine how robust GC B cell responses to influenza vaccination in humans could overcome antigenic imprinting and if that correlates with a more durable antibody response. Finally, we will genetically analyze the mechanisms of action of the transcriptional determinants that are preferentially expressed in plasma cells destined for longevity. Our findings will potentially reveal the cellular and molecular determinants dictating the longevity and the breadth of elicited antibody responses to influenza – and potentially other – vaccination in humans.

NIH Research Projects · FY 2025 · 2018-09

The scientific premise of this application is that the individualized translational research process of the Undiagnosed Diseases Network (UDN) is sustainable and that its impact on patients, families, and disease discovery can be further extended by increasing its accessibility to a broader participant population. This application would support the Washington University in St. Louis (WashU) UDN Diagnostic Center of Excellence (DCoE). WashU and BJC Healthcare together represent a large academic medical center, hospital, and outpatient clinical system that is fully integrated with world-renowned basic science capabilities and demonstrated expertise in finding diagnoses for patients with undiagnosed diseases. The highly collaborative clinical and biomedical research culture at WashU promotes interactions within and across Departments, with institutional genomic, clinical, computational, and model system experts, and with colleagues regionally, nationally, and internationally. These interactions support recruitment, selection, evaluation, diagnosis discovery, and follow up of pediatric and adult patients with undiagnosed diseases through both established networks and individual referrals. Building on this infrastructure, we propose to continue to advance the success of the UDN in diagnosing and managing disease in undiagnosed patients by utilizing a system for clinical and genomic review of applications and participant cases, with expanded, expert clinical, laboratory, and informatics expertise to solve cases that are and are not suspected to have genetic underpinnings. Specific strengths of our DCoE include neurological disorders (neurodevelopmental, neuromuscle), inborn errors of immunity, inherited pulmonary disorders, and disorders caused by somatic mosaicism. Second, we aim to improve access to the UDN for participants in the community. In particular, we will focus our outreach program through a Community Engagement Strategy that includes establishment of a collaboration with healthcare organization supporting federally-qualified health centers and the Saint Louis County Health Department, with appointment of a new Community Engagement Officer. Third, we will implement a Sustainability Strategy including research support, insurance billing, charitable funds, and departmental cost-sharing. We will work closely with the UDN Network and the Data Management and Coordinating Center to implement collaborative, highly effective, equitable, and sustainable methods for undiagnosed patients.

NIH Research Projects · FY 2024 · 2018-09

Abstract Down syndrome (DS), the most common genetic cause of intellectual disability, is associated with varying degrees of cognitive and behavioral impairments. Pharmacologic therapies and genetic modulators are emerging which, if administered early in conjunction with traditional therapies, show promise for improving developmental outcomes in children with DS. However, the stark absence of early neurodevelopment knowledge in DS hampers these efforts. The main goal of this proposal is to develop biomarkers as future targets for specific therapies based on careful characterization of early aberrant neurodevelopmental patterns. This study will be a combined effort with WU as the coordinating center and six other research groups: UNC, UW, CHOP, MNI, NYU, and University of Minnesota. These groups comprise the ACE-IBIS (Autism Center of Excellence Infant Brain Imaging Study) network, a well-established and experienced group that has been productively collaborating for 8 years on MRI imaging and behavioral characterization of infants at high risk for autism, healthy typical infants (TYP), and infants with Fragile X. The aims of this proposal are: 1) Define the longitudinal characteristics of early brain development in infants (3 to 24 months) with DS in comparison to TYP infants and infants with other developmental disabilities (ASD and Fragile X) using three different types of neuroimaging (MRI, DTI, rsfMRI); 2) Develop predictive models for developmental outcomes in infants with DS based on longitudinal structural or functional MRI characteristics; and 3) Characterize brain-behavior correlates with coordinated multimodal imaging in infancy characterizing the interrelationship between longitudinal network imaging parameters and cognitive, behavioral and neurodevelopmental outcomes using sophisticated multivariate support vector machine (SVM) analytic strategies. 120 infants with DS and 40 TYP control infants will be followed longitudinally from 3 to 24 months. MRI scans will be obtained during natural sleep and a series of well-validated developmental and behavioral assessments will be completed at each visit. This project will be the first to define the nature and timing of alterations in brain development in infants with DS. The proposed project addresses several key research recommendations from the “NICHD 2014-The NIH Research Plan on Down Syndrome.” The study aims match the recommendation for the quantitative characterization based on imaging of early brain development and the relationship of cognitive, behavioral and social development to early aberrant neurodevelopment in DS. This project will also address recommendations for investigation of comorbid ASD in DS, which could be as high as 5-10%. The IBIS network is uniquely qualified to examine early neurodevelopmental patterns, utilizing the ASD, TYP and FraX infant data sets to better characterize and examine specificity of DS early developmental patterns including ASD qualities and impairments in social development. It is clear that quantification of neurodevelopmental trajectories in infants with DS will be critical to identification of personalized intervention strategies and to assess the efficacy of early life, targeted, highly novel and mechanistically specific DS interventions.

NIH Research Projects · FY 2025 · 2018-09

Abstract This proposal was specifically designed to support a dedicated Research Specialist with extensive experience in human induced pluripotent stem cell (hiPSC) engineering to study the cellular and molecular mechanisms underlying the formation and progression of central and peripheral nervous system tumors relevant to the identification of risk assessment and personalized therapeutic strategies. This investigator has longstanding expertise in RASopathy cancer predisposition syndromes, beginning with her doctoral studies focused on generating Cardio-Facial-Cutaneous and Costello Syndrome zebrafish preclinical models, and over the past ten years, initially as a postdoctoral fellow and now as an Assistant Professor on the Research track, in developing translational research tools for the Neurofibromatosis Type 1 (NF1) cancer predisposition syndrome. As such, the Research Specialist established and continues to expand the Washington University NF Center hiPSC repository, which includes hiPSCs with patient-derived germline mutations, as well as engineered tagged or conditional knockout lines, explicitly designed to define the molecular and cellular etiologies underlying NF1- and RAS/MEK-associated nervous system cancers in children and adults. Leveraging the power of hiPSC engineering, this Research Specialist has pioneered work demonstrating that (1) NF1 mutations are not functionally equivalent in vitro or in vivo (cancer genetics), (2) NF1-mutant neurons drive low-grade nervous system tumor growth in an activity-dependent manner (cancer neuroscience), (3) neurons function as immune activators to establish an immune cell axis supportive of brain tumor growth (cancer immunology), (4) targeting neuronal excitability provides new options for the treatment of brain and nerve tumors (cancer therapeutics), (5) hiPSCs can be exploited to develop novel humanized models of low-grade brain and nerve tumors (cancer modeling), and (6) hiPSC cancer modeling revealed a chemokine circuit critical for patient-derived low-grade brain tumor xenograft generation (precision oncology). Moreover, multi-institutional international dissemination of these unique tools has galvanized new research directions within the Program Director’s laboratory, critical to the successful acquisition of new NCI grant funding and the establishment of new international collaborations. Finally, the Research Specialist’s commitment to the field, continued leadership in this scientific area, and critical importance in guiding hiPSC-based projects, both at Washington University and worldwide, will continue to provide clinically-actionable opportunities relevant to personalized risk assessment and medical management of people with RAS-related oncologic disease.

NIH Research Projects · FY 2025 · 2018-09

PROJECT SUMMARY The DIAN-TU platform was formed to design and manage interventional therapeutic trials and find a treatment that provides cognitive benefit for those certain to develop dominantly inherited AD (DIAD). The DIAN-TU trial platform is now fully operational in 6 countries and 24 sites with another 13 countries and 26 sites in start-up. The current DIAN-TU-001 trial will accommodate 11 languages and has three different therapies being tested in secondary prevention (i.e. cognitively normal participants with substantial AD pathology). NIA funding for the DIAN-TU trial platform established the infrastructure and operations for executing clinical trials in DIAD and acknowledged the need for evolution within this platform. The DIAN-TU Primary Prevention Trial is a first of its kind, phase II/III, 4-year randomized, blinded placebo-controlled (1:1) trial in 160 asymptomatic dominantly inherited Alzheimer disease mutation carriers who are more than 15 years before the estimated year of symptom onset (EYO) and have minimal to no Aß- PiB plaque burden at trial entry. Current trials in asymptomatic individuals target Aß after pathology is established; these secondary prevention efforts are likely more effective than treating at later more advanced stages, however the most effective approach is to prevent AD pathology from forming. The goal of this proposal is to implement a placebo controlled biomarker endpoint clinical trial targeting amyloid deposition in subjects at risk for DIAD, prior to onset of significant Aß pathology. In this study, we will test if it is possible to prevent Aß deposition in DIAD mutation carriers and if doing so will prevent the cascade of pathology associated with AD and, ultimately, dementia in a population that is otherwise certain to get the disease. Regardless of the outcome of this study, it will be highly impactful on the AD field in assessing the ability to prevent amyloidosis and the consequences of doing so at the earliest stages of the AD pathological cascade. If the prevention of Aß pathology in DIAD is accomplished, it will lay the foundation for the ultimate test of the amyloid hypothesis and provide the best opportunity to prove that dementia in this highly vulnerable population, and possibly in sporadic AD and Down syndrome, can be dramatically modified. Should preventing amyloid pathology from developing have no impact on the course of the disease, particularly in this population, this would direct future research and therapeutics towards other mechanisms and pathologies.

NIH Research Projects · FY 2026 · 2018-09

PROJECT SUMMARY The DIAN-TU platform was formed to design and manage interventional therapeutic trials and find a treatment that provides cognitive benefit for those certain to develop dominantly inherited AD (DIAD). The DIAN-TU trial platform is now fully operational in 16 countries and 40 sites and accommodates 11 languages. NIA funding for the DIAN-TU trial platform established the infrastructure and operations for executing clinical trials in DIAD and acknowledged the need for evolution within this platform. The DIAN-TU Primary Prevention trial will lay the foundation for the ultimate test of the amyloid hypothesis and provide the best opportunity to prove that dementia in this highly vulnerable population, and possibly in sporadic AD, can be dramatically modified, if not prevented altogether. The DIAN-TU Primary Prevention Trial is a first of its kind, phase II/III, multi-stage randomized, blinded placebo-controlled (1:1) trial in 160 asymptomatic dominantly inherited Alzheimer disease mutation carriers who are 11 to 25 years before the estimated year of symptom onset (EYO) and have minimal to no Aß-PiB plaque burden at trial entry. The first therapy tested will be with remternetug. The recent findings in the field for lecanemab and donanemab have demonstrated that removal of amyloid plaques can reduce cognitive decline, reinforcing the need to test treatment before amyloid begins to accumulate in the brain, i.e. primary prevention. Directly testing the amyloid hypothesis of AD and the potential impact the outcomes could have on the overall burden of Alzheimer's disease (AD) in the U.S., this project addresses a major public health problem. In this study, we will test if it is possible to prevent Aß deposition in DIAD mutation carriers and if doing so will prevent the cascade of pathology associated with AD and, ultimately, dementia in a population that is otherwise certain to get the disease. The results of this study are likely to be highly impactful on the AD field in assessing the ability to prevent amyloidosis and the consequences of doing so at the earliest stages of the AD pathological cascade. If the prevention of Aß pathology in DIAD is accomplished, it will provide one of the best scientific tests of the amyloid hypothesis and provide the best opportunity to prove that dementia in this highly vulnerable population, and possibly in sporadic AD and Down syndrome, can be dramatically modified. Should preventing amyloid pathology from developing have no impact on the course of the disease, particularly in this population, this would direct future research and therapeutics towards other mechanisms and pathologies.

NIH Research Projects · FY 2025 · 2018-09

This competing renewal, “Strengthening Child Health Research Capacity in Resource Constrained Settings” training program (referred to hereafter as” the training program”) aims to advance and test state-of-the-art research methods training and “hands-on” research experience for advanced doctoral students and early career investigators, committed to addressing the serious threats to child health, as well as prevention and care differences in poverty-impacted contexts. The training program develops and supports a pipeline of new child health research investigators who are prepared to advance scientific knowledge about system and community-level structural interventions that can address the disproportionate health burdens experienced by poverty-impacted youth via enhancing protective family, neighborhood, system supports; reducing differences; and advancing health access and availability. The training program is guided by 4 Specific Aims: Aim 1. Recruit 5 cohorts of advanced doctoral students and early career investigators, committed to conducting child health and behavioral health prevention, intervention, services, implementation, and scale-up research within resource-constrained settings (Fellows; n=45 across 5 years); Aim 2. Deliver a summer research training program aimed at equipping Fellows with skills to address the challenges in resource-poor settings through didactic instruction, mentoring, “hands-on” immersion in child- and family-focused studies, individualized consultation, goal setting, monitoring, and ongoing support in resources over time; Aim 3. Advance academic/community/safety net system research partnerships on child health and well-being; and Aim 4. Examine the short-term and longitudinal impact of the training program (across 10 cohorts; n=90 RRT Fellows). Fellows participate in a 2-week, face-to-face training program at Washington University in St. Louis. Fellows then spend 4 to 6 weeks embedded across a set of existing child health-focused research studies. A rigorous mixed-methods evaluation tracks individual Fellow progress, as well as the impact of the training program on overall child health research partnerships.

NIH Research Projects · FY 2025 · 2018-09

PROJECT SUMMARY Almost all cellular organisms employ an array of photoreceptors to detect their light environment. Arguably the most influential are the phytochromes (Phys), a diverse group essential for plant development and the behavior of many bacterial, fungal, and algal species. By reversible photointerconversion of their bilin (or open-chain tetrapyrrole) chromophores between red light-absorbing Pr and far-red light-absorbing Pfr states, Phys act as photoswitches in various signaling cascades responsive to light intensity, duration, direction, and spectral quality. Moreover, through the thermal reversion of Pfr back to Pr, some Phys sense temperature through enthalpic effects on this reaction, and perceive photoperiod through the nighttime depletion of Pfr. The cumulative effects of this Pr/Pfr interconversion impact numerous physiological processes important to agriculture and the biology of harmful plant and human pathogens. In addition, their unique photochemistries provide invaluable optogenetic tools, including novel fluorophores for tissue imaging, and engineered photoswitches for regulating cellular events with remarkable temporal and spatial precision. Recently, we made great strides in understanding how Phys signal, with emerging structures suggesting that microbial and plant Phys use two distinct output modalities. Both start with light-triggered isomerization of the bilin, which drives a - stranded to -helical rearrangement of a hairpin loop that links the signature PHY and GAF domains. While photoactivated microbial Phys then connect torsional strain generated within the dimer to regulate an appended output domain (typically with histidine kinase activity), plant Phys have rearranged their domain organization to create a photosensitive dimeric platform that likely enables reversible binding and eventual degradation of the family of PIF transcriptional repressors. While current models helped illuminate gross changes required for endstate conversion, the intermediates of photoexcitation and ensuing structural changes necessary for creating a signaling-competent Pfr state remain uncertain. The objectives of this proposal are to complete these pictures through continued X-ray crystallographic and cryo-electron microscopic approaches followed by informed biochemical analyses of representatives in their inactive and active states. Specific aims are to: (1) exploit time-resolved serial X-ray crystallography to structurally define the intermediates generated by Phys after photon absorption; (2) generate more comprehensive structures of bacterial Phys, including models of full- length dimeric photoreceptors with their signal output modules; (3) develop a model for how plant Phys signal through structural studies on Pfr; (4) apply steady-state and time-resolved protein surface mapping to support the Phy photoconversion pathway(s) seen structurally; (5) develop models of Phys interacting with their downstream effectors, and (6) appreciate how diversity among plant Phy enables thermal/time perception by specific isoforms. Taken together, this project will provide an essential framework to better appreciate the structure, allosteric mechanisms, and evolution of the Phy superfamily. Understanding how microorganisms and plants sense light, temperature, and time would then have important ramifications for improving the agricultural performance of crop plants, understanding microbial ecosystems, controlling medically-relevant pathogens, and furthering the application of Phys as optogenetic reagents.

NIH Research Projects · FY 2025 · 2018-09

The WU-RDRRC seeks to advance the health of patients with rheumatic diseases by supporting enabling technology and promoting the members’ basic, clinical, and translational research interests that are organized around three major themes: 1) elucidating basic mechanisms of inflammation and autoimmunity; 2) accelerating clinical/translational research to inform precision medicine; and 3) advancing genome engineering and regenerative medicine to develop new treatment approaches for rheumatic diseases. We posit that translational research endeavors in rheumatic diseases require a team approach cultivated in a vibrant environment and supported by cross-disciplinary groups of experts and cutting-edge technologies. Over the past four years, this operational philosophy and the infrastructure supported by the WU-RDRRC perfectly positioned our investigators to increase the efficiency and impact of their research, allowed them to quickly address emergent issues that arose during the COVID-19 pandemic, and helped a number of young investigators obtain extramural funding and successfully transition to research independence. The WU-RDRRC propose to continue promoting rheumatic disease research by: 1) providing the infrastructure, education, and training needed to assist investigators with the experimental and scientific design of their projects; 2) organizing and supporting Core laboratories that will adopt and apply innovative technologies and expertise that would otherwise be unavailable to investigators in a cost-effective manner; 3) promoting interest in rheumatic diseases research by engaging multidisciplinary teams and bringing together research disciplines and clinical divisions to address roadblocks in translational science; 4) fostering the next generation of junior investigators who are interested in the study of rheumatic disease-related areas. To accomplish our goals we propose four cores: 1) a Biobank and Phenotyping Core to collect high-quality biospecimens for downstream molecular analysis; 2) a Genome Engineering Core to generate bioengineered cell-based therapies, knockin/knockout mice for testing in preclinical disease models; 3) a Cellular Imaging Core that offers novel microscopy technologies for building models of health and disease; and 4) an Administrative Core that promotes collaborative and synergistic interactions among rheumatic disease researchers and the mentoring of junior and new investigators interested in rheumatic disease research.*

NIH Research Projects · FY 2025 · 2018-09

Overall Project Summary In this renewal application of our National Center for Biomedical Imaging and Bioengineering (NCBIB), the PET Radiotracer and Translation Center (PET-RTRC), our objective is to further fortify the Center as a national resource that leverages the expertise at Washington University and the Mallinckrodt Institute of Radiology in PET radiotracer design, development, production, training and dissemination. In collaboration with research groups throughout the country and in Europe who are studying molecular and cellular processes of inflammation in disease, we will further enhance the development and dissemination of novel PET radiotracers needed to transform biomedical research and advance human health. In the process we will strengthen the foundation for propelling the PET-RTRC forward and sustaining its long term operation. To fulfill our objective we will address the following Specific Aims: Aim 1: In responding to the scientific needs of the Collaborative Projects (CPs), the three Technological Research & Development Projects (TR&Ds 1-3) will build upon their successful innovations during the current funding cycle to develop radiotracers that target different components of inflammation, their dynamic variation, and consequences such as inflammasome activation (TR&D 1: sphingosine-1-phosphate receptor-2), monocyte/macrophage trafficking (TR&D 2: chemokine C receptor 2/CD163) and the induction of mitochondrial stress (TR&D 3: mitochondrial reactive oxygen species). Aim 2: Given the importance of the CPs as technology drivers, we will further enhance the “push-pull” interaction between the individual TR&Ds and their respective CPs by including strategies that enhance cross-site communication, more robust training opportunities, optimized image analysis and data management and facilitated sharing of intellectual property. Aim 3: The Technology Training and Dissemination Core expands its success in the current grant cycle to offer robust training and dissemination opportunities. In the training component, all Center workshops and seminars will be presented in a hybrid onsite/remote mode, individualized training will be offered via proctored “how to” videos on key topics and additional outreach initiatives will be implemented that are geared towards interactions with other P41 Centers, internal WU training grants/programs and traditionally underserved universities. New innovations in our dissemination efforts will increase the capabilities of our Service Projects for both pre-clinical and human studies. The Administration Core will continue to provide the managerial oversight necessary for the efficient operation of the PET-RTRC so that it meets its scientific, training and dissemination objectives. Successful completion of the proposed renewal application will further propel the PET-RTRC as a national resource to facilitate research that will increase our understanding of the molecular basis of disease, thus providing the framework for novel diagnostic and therapeutic paradigms leading to improved human health.

NIH Research Projects · FY 2026 · 2018-08

ABSTRACT Stroke is a major public health problem in the United States, where it is the 5th leading cause of death and the leading cause of adult disability. There is great need for translational and clinical research to develop interventions that prevent, treat, or enhance recovery after stroke. However, clinical trials require significant infrastructure, and the time and energy required to assemble this infrastructure for individual trials is a major barrier to progress. The NINDS Stroke Net was established in 2013 (we joined in 2018) as a national stroke trials infrastructure, designed to maximize efficiencies by centralizing regulatory and contractual agreements. With a single National Clinical Coordinating Center (NCC), a single National Data Management Center (DMC), and 29 Regional Coordinating Centers (RCCs) throughout the US, Stroke Net was established to efficiently recruit patients for stroke trials. The existing Stroke Net RCC28 Mid-America Regional Coordinating Center (MARCC) at Washington University School of Medicine (WU) and affiliated Barnes-Jewish Hospital (BJH), in collaboration with five other high-volume tertiary care Stroke Centers spans the Midwestern and Southern states of MO, KS, IL, TN, AR: St. Luke’s Hospital in Kansas City, MO; The Order of St. Francis Medical Center (OSF) in Peoria, IL; University of Tennessee Health Sciences in Memphis, TN; Cox Medical Center in Springfield, MO; and the University of Missouri in Colombia, MO. While not heavily represented in NIH clinical trials, these states have some of the highest stroke prevalence in the US. Missouri is ranked 19th in stroke mortality, Tennessee ranked 8th, Mississippi ranks 1st, Illinois 15th and Kansas 27th among all states according to 2020 CDC data. Since our last submission in 2018, the Mid-America Regional Coordinating Center (MARCC) at Washington University School of Medicine (WU) and affiliated Barnes-Jewish Hospital (BJH), has grown through enhanced attrition of low performing sites and addition of new sites including a large academic hub and five satellite high-volume tertiary care Stroke Centers. Each center has been carefully selected to serve as a tertiary care hub with large networks of referring hospitals, and has extensive experience in multi-center stroke trials. We consider MARCC a “Network of Networks”. Drawing from both rural and urban settings, the hospitals of MARCC admit over 7,500 stroke patients per year, including patients from diverse racial and socioeconomic backgrounds. Moreover, the MARCC collective group of stroke clinicians, investigators, coordinators, and scientists has successfully conducted Stroke Net trials, ranking 4th in overall Stroke Net Subject Enrollment from 2018-23 and ranking 4th in overall enrollment of Black Americans. We propose that MARCC will continue to perform as one of Stroke Net’s highest performing RCCs with the following specific aims: Aim 1. To maintain a seamless multi-state stroke trials infrastructure—the Mid-America Regional Coordinating Center (MARCC). Aim 2. To lead NINDS Stroke Net enrollment of patients from diverse racial and socioeconomic backgrounds. Aim 3. To recruit and train future clinician-scientists in stroke research through fellowship training and career enhancement.

NIH Research Projects · FY 2026 · 2018-08

PROJECT SUMMARY A delicate regulatory balance must be achieved in cells of the innate and adaptive immune systems to effectively eliminate pathogens, while minimizing damage in neighboring tissues. Defects in regulatory mechanisms that govern expression of cellular or soluble mediators can interfere with pathogen clearance or lead to unchecked inflammatory responses associated with autoimmunity. We now appreciate that the innate immune system includes functional counterparts of T helper (Th) cells, but the innate cells lack antigen-specific receptors and respond with enhanced kinetics and vigor to danger signals induced by pathogenic insults. The Th counterparts, called innate lymphoid cells (ILCs), also have been implicated in the pathogenesis of several autoimmune diseases, including inflammatory bowel disease (IBD). In discovery-driven profiling studies, the MPIs have defined chromatin landscapes of Th-ILC counterparts derived from human mucosae, revealing collections of regulatory elements (REs) that may control the expression of key immune mediators. Moreover, many REs that were active in specific ILC or Th subsets co-localized with disease-associated SNPs, and are conserved in mice, suggesting they may be important for controlling expression levels of nearby genes in inflammation-driven pathogenesis. Indeed, the MPIs have defined novel functions for two such REs in controlling expression of a signature cytokine (IL-22) and a gene required for effector functions in most lymphoid cells (Tcf7). Despite this progress, the roles of potentially important REs in cell type-, agonist-, and disease-specific gene expression largely remain untested. The goal of the current project is to address these outstanding issues, focusing on regulation of a broad panel of signature genes that control ILC-Th function. To achieve these goals, we will leverage the MPIs’ complementary expertise. Dr. Colonna’s lab discovered several ILC subsets and continues to contribute to our understanding of their biology in mice and humans. Dr. Oltz’s lab studies cis-regulatory circuits that drive lymphocyte development and transformation. Two specific aims are proposed to test the hypotheses that: (i) unique sets of REs are critical for cell type- and agonist- specific expression of ILC3-Th17/22 cell signature genes in health and disease, (ii) the transcription factors ZFP683 and PRDM1 orchestrate key aspects of regulomes that govern the biology of natural killer (NK) cells and their ILC1 counterparts. Continuation of our productive collaboration will identify and functionally test critical players – both REs and transcription factors – that dominantly regulate expression patterns of genes involved in inflammatory diseases, providing insights into independent roles of cytokine expressing cells in pathogenesis, and ultimately opening new therapeutic avenues.

NIH Research Projects · FY 2026 · 2018-08

Abstract Interstitial cystitis/Bladder Pain Syndrome (IC/BPS) is a serious and painful condition of unknown etiology that affects 6% of women in the United States. The major clinical symptoms of IC/BPS are pain on bladder filling and increased urinary urgency and frequency. The majority of IC/BPS patients (90%) also suffer from comorbid anxiety and/or depression, contributing to a poor quality of life. Current available treatments for IC/BPS are largely ineffective, providing only mild symptomatic relief. Given the prevalence of this debilitating disease and the lack of effective treatments, further studies are needed to better understand the underlying mechanisms that contribute IC/BPS and those that mediate the high incidence of comorbid anxiety and depression. We previously reported that IC/BPS patients show referred abdominal hyperalgesia, a clear sign of central sensitization. In the previous funding period, we used the single nucleus RNA sequencing, spatial transcriptomics and in situ hybridization and identified two populations of CeA neurons that are activated in cystitis and appear to play opposing roles in the regulation of bladder pain. Here we will apply state of the art approaches in systems neuroscience to unravel the potential role of these unique neuronal subpopulations in the CeA in the reciprocal regulation of pain, voiding dysfunction, and negative affective behaviors (IC/BPS-like conditions). What are the respective roles of the two populations in bladder pain? Are the apparent pro- and anti-nociceptive actions of the Pde1c and Cartpt populations, respectively, restricted to referred hypersensitivity, or do they also reciprocally regulate ongoing pain? Do these populations play differential roles in the regulation of voiding behavior and negative affective behaviors? What are the critical inputs to and projections from these neurons? Do these populations undergo plasticity differentially in the induction and maintenance of cystitis? How do the dynamics of this circuit change as cystitis resolves? Here we propose to answer these questions in a series of studies that test the central hypothesis that maladaptive plasticity in these unique subpopulations of CeA neurons regulates voiding dysfunction, pain sensitization, and comorbid negative affect in models of cystitis. These studies will provide new insights into the critical role of the pro- and anti-nociceptive neurons in the CeA in bladder pain and comorbid affective disorders in the context of bladder pain syndrome. If successful, these studies will point the way to identifying pharmacological approaches to restore normal circuit function around these neurons to provide relief from bladder pain, voiding dysfunction, and comorbid anxiety and depression in patients with IC/BPS.

NIH Research Projects · FY 2026 · 2018-07

The identification of the association of the TREM2 p.R47H varienat with Alzheimer's disease (AD) risk highlighted the impact of the immune response and inflammation in (AD). However, the role of TREM2 in health and in AD as well as what other genes are part of the TREM2 pathway are not totally known. We have demonstrated that soluble TREM2 (sTREM2) levels may reflect biological events that link amyloid deposition and neurofibrillary tangle formation to cognitive decline, and that can be used to genetic studies. Using CSF sTREM2 levels as endophenotype we demonstrated that MS4A4A is the major regular of sTREM2 and a potential target. In a more recent study in almost 3,000 CSF samples, we identified four loci, including MS4A4A, TREM2, TGFBR2 and NECTIN2. In this proposal, we will perform the first multi-tissue genetic screening of sTREM2 regulators. We will analyze very large datasets in brain (n=5,164), CSF (n=6,800) and plasma (n=56,900) from well-characterized cohorts with extensive pre-existing CSF biomarker (Aβ, Tau, ptau and sTREM2), clinical and genetic data. The aims of the project are: 1) to identify single (common and rare) variants, genes and pathways associated with sTREM2 levels; 2) to identify multi-tissue multi-omic signatures of TREM2 risk variants and 3) to perform functional analyses in iPSC-derived human microglia to determine the mechanisms by which TGFBR2 and NECTIN2 affect sTREM2 levels and microglia function.

NIH Research Projects · FY 2025 · 2018-07

The overarching goal of this K24 application is to further my development as a clinical investigator and support the training of future physician scientists. My patient oriented research interests relate to understanding the clinical, pathologic, genetic and pathomechanistic underpinnings of inherited and acquired forms of muscle weakness. With the support of this grant, I will continue to perform genetic discovery of patients with muscle disease, increase our biorepository, resolve variants of unknown significance and understand the phenotypic spectrum of these diseases. In addition, I will increase my mentorship responsibilities of graduate students, resident physicians, fellows and junior faculty within the neuromuscular group and Department of Neurology at Washington University School of Medicine. A K24 grant would protect 50% effort and relieve future clinical and administrative responsibilities. The two interrelated aims of this proposal are 1) Resolve variants of unknown significance in LGMD genes. 2) Perform natural history studies related to rare muscle diseases. These aims will be achieved utilizing our existing biorepository within the Washington University School of Medicine Neuromuscular Genetics Project and the acquisition of new patients and patient material. Support through a K24 Midcareer Investigator Award in Patient-Oriented Research would come at a critical time in my career as I solidify my independent research program and increase my availability to mentor graduate students, post-doctoral fellows, residents, neuromuscular fellows and junior faculty in translational myology. Upon completion of this award, I will have integrated clinical trainees into our existing translational research infrastructure and created a successful pipeline to generate the next generation of clinician- scientists focused on muscle diseases.

NIH Research Projects · FY 2026 · 2018-06

ABSTRACT Substantial investments are being made to sequence the genomes of families with autism and other neurodevelopmental disorders (NDD). However, identifying disease mutations outside the ~1% of protein coding sequences is challenging because 1) the ‘search space’ is much larger, and thus many more mutations occur by chance, and 2) there is no simple code to identify deleterious mutations in non-coding sequences, and thus loss of function mutations must be defined experimentally. In addition, the consequences of mutations in non-coding (i.e. regulatory) sequences are often highly dependent on the specific cell type. Thus, functional assays must be conducted in vivo, in the appropriate CNS cell types. To address the search space challenge, we have focused specifically on the untranslated regions (UTRs) of mRNAs. UTRs are important, conserved regulatory sequences that profoundly impact protein levels by altering translation rates or transcript stability for specific genes. To address the lack of a code for interpreting UTR mutations, we have developed a unique combination of expertise to conduct massively parallel functional analysis of UTR variants from NDD patients, in relevant cell types in the brain. Combining two innovative but established components: massively parallel reporter assays, and cell type specific translational profiling, we aim to establish a pipeline to 1) Identify UTR mutations that result in altered protein levels, 2) conduct genetic burden and association testing on these variants, and 3) train machine learning models that can predict the effects of future mutations in developing neurons of the brain. This project will leverage the existing large investment in NDD genome sequencing by defining individual non-coding, disease- causing mutations in a class of sequences that has, so far, not been the focus of disease studies.

NIH Research Projects · FY 2026 · 2018-06

Project Summary The fibers of the ciliary zonule suspend the lens on the optical axis and transmit the forces that flatten it during disaccommodation. Mutations in genes encoding zonular proteins underlie syndromic and non-syndromic conditions that affect the eye profoundly. Common ocular phenotypes include ectopia lentis (lens dislocation), cataract, axial elongation, myopia, glaucoma, and retinal detachment. The molecular composition of the zonule was recently elucidated, but the mechanism by which mutations in zonular components culminate in structural failure of the fibers is unknown. In Aim 1, therefore, three zonulopathies (Marfan Syndrome, Weill-Marchesani Syndrome, and Isolated Ectopia Lentis) will be modeled in mice. Utilizing recently developed imaging and material testing techniques, we will examine how, in each case, the structure and viscoelastic properties of the mouse zonule are affected by the presence of the mutant protein (or absence of the wild-type protein). We hypothesize that the initial pressurization of the eye is a critical step in zonule development. In Aim 2, this notion will be tested by measuring the rise in intraocular pressure in postnatal mice and determining whether pressurization of the developing eye in vitro causes precocious deployment of zonular fibers. Preliminary studies identified the cross-linking enzyme lysyl oxidase-like-1 (LOXL1) as an abundant component of the zonule proteome. In Aim 3, we propose that LOXL1-derived cross-links have a critical role in strengthening the zonule. We will test that hypothesis in a knockout mouse model. Finally, microspherophakia (i.e., the presence of a smaller and more spherical lens) is observed in Weill-Marchesani patients (who harbor mutations in LTBP2 or FBN1, zonular proteins that contribute to the tensile properties of the fibers). We hypothesize that forces exerted by the zonular fibers on the lens surface influence lens growth. In Aim 4, we will elucidate the three dimensional structure of the human zonule and correlate the distribution of proliferating lens epithelial cells with the strain fields established around zonular attachment points.

NIH Research Projects · FY 2026 · 2018-05

PROJECT SUMMARY Early exposure to social disadvantage, specifically family and neighborhood financial disadvantage in utero and through the first 3 years of life, is a powerful and potentially modifiable environmental determinant of risk for childhood psychopathology. The ongoing Early Life Adversity and Biological Embedding (eLABE) study launched in 2017 has aimed to elucidate mechanisms of this risk trajectory and guide preventive interventions. To date, eLABE has detected powerful associations between prenatal social disadvantage and structural and functional brain development at birth which are mediated through elevations in maternal cytokines and alterations in the gut microbiome. These neonatal neurodevelopmental variations have been linked to elevations in markers of psychopathology risk at age 2 years. Further, decreased caregiver nurturance mediated some aspects of this relationship, with supportive caregiving mitigating child psychopathology risk. Our central hypothesis is that pro- inflammatory immune and gut microbiome profiles evident in children exposed to prenatal and early life social disadvantage induce neuronal effects that negatively impact structural and functional brain development leading to increased risk for psychopathology. This renewal proposes to build upon the existing data to continue following our unique and well-characterized sample of 377 children for whom we have assembled a rich, unparalleled repository including comprehensive measures of social disadvantage (in utero and ages 1, 2, and 3 years) with early life measures of inflammation, the gut microbiome, and brain development (structural, microstructural, and functional MRI at birth and ages 2 and 3). In the next phase of eLABE, we propose to follow this sample into school entry (age 6) and middle childhood (age 8), adding assessments of social disadvantage, developmental/behavioral, inflammatory marker, multimodal MRI, and caregiver and social support measures at both timepoints and one additional wave of the gut microbiome at age 6. We will utilize these data to test specific hypotheses about mechanistic pathways to mental disorders that begin to clearly emerge in early childhood and school age, and will utilize novel brain metrics to more directly examine the roles of neuroinflammation and plasticity in this risk pathway. We will also examine the effects of caregiver and family, peer, and school supports on this risk trajectory, including their role as possible resilience factors, and explore whether there are sensitive periods for these effects. The results will provide the first systematic and intensive prospective examination of the relationship of social disadvantage to chronic systemic inflammation, the gut microbiome, and their links to brain development and behavior related to risk for clinical mental disorders. Critically, by targeting caregiving, inflammation, and the gut microbiome as key mechanistic pathways, these data will also have tremendous clinical and translational potential as they investigate potential mediators of risk and provide modifiable targets for prevention of mental disorders.

NIH Research Projects · FY 2026 · 2018-05

Summary Poor understanding of the mechanisms that modulate virus pathogenesis limits our ability to control viral infections and reduce their public health burden. Non-standard viral genomes of the copy-back type (cbVGs) that are generated during viral infections are the primary inducers of antiviral immunity to respiratory syncytial virus (RSV) in vitro, in mice, and in humans. Most importantly, we recently reported that cbVGs generated during RSV replication significantly impact the clinical outcome of infection in children and adults. These data suggest that cbVGs are key determinants of viral pathogenesis and that their activity can be harnessed to minimize viral-associated disease. It's clear that the virus host-interaction is complex, and it is essential to untangle this complexity to identify better predictors of disease severity as well as better therapeutic strategies. A better understanding of the factors that influence the generation and activity of cbVGs is necessary for exploiting them as a tool for reducing the public health burden of RSV and related viruses. In studies funded in the original proposal, we demonstrated that the presence of cbVG critically impact RSV pathogenesis. While detection of cbVGs soon after infection is protective from severe disease, late or sustained presence of cbVGs associates with more severe disease. Studies proposed here, directly follow these data and focus on investigating viral and host factors that impact cbVG generation and activity. To do this, we will characterize the cbVG species present in human respiratory secretions and we will study their function and association with distinct clinical outcomes. This study will provide the first comprehensive investigation of naturally occurring cbVG species and will identify unique features that determine their protective or potentially pathogenic functions (Aim 1). We will also assess the impact of known RSV risk factors on cbVG generation to identify host determinants of cbVG accumulation, and we will perform proof of concept experiments to test if cbVGs can be safely used to minimize virus-induced disease in high-risk settings (Aim 2). Lastly, we will test the role of the host immune bias in driving disease in patients with late or prolonged cbVGs and will identify potential targets for treatment to reduce disease in these conditions (Aim 3).