Washington University

NIH Research Projects · FY 2026 · 2023-02

Project Summary As animals locomote, they constantly coordinate activity between the rostral and caudal ends of the body. This coordination relies on neurons in the spinal cord that send long ascending or descending axonal projections. Although several studies have shown that ablation of these neurons leads to deficits in coordination, it is largely unknown whether these long-range circuits are similar or different from local circuits. In recently published data in larval zebrafish, we demonstrated that at least one genetically defined class of spinal neurons, the V1 (En1+) population, changes its synaptic targets as its axon ascends in the spinal cord. Specifically, V1 neurons form synaptic connections with motor neurons and other ventral horn neurons nearby, but switch to inhibiting a dorsal horn sensory population at longer range. Here we propose to extend this analysis to five additional sets of ventral horn neurons, the dI6, V0 excitatory and inhibitory, V2a, and V2b populations. To create this large-scale circuit map, we use localized optogenetic activation of identified spinal populations while carrying out whole-cell recording of identified potential postsynaptic partners. By translating the optogenetic stimuli up and down the spinal cord, we can build a physiological map of the strength of the synaptic connection at various rostrocaudal positions. Normalization of the synaptic charge transfer allows comparisons across target populations, providing a comprehensive grid of connectivity among spinal neuron classes at various rostrocaudal distances. We will then build a computational model of spinal cord connectivity that reflects biological reality, as measured in these experiments. Using this model, we will test the consequences of shifting synaptic connections in the rostrocaudal axis, in order to understand the logic of spinal circuit organization. Finally, we will selectively ablate long-range or local V2a neurons to determine the behavioral effects of long-range vs local projections. Together, these experiments will provide a circuit map of genetically identified spinal neurons.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY / ABSTRACT Antimicrobial resistance (AMR) contributes to an estimated 5 million deaths worldwide each year and is directly responsible for over 1.2 million deaths. In the not-to-distant future, we may face a reality where infections resistant to all existing antibiotics are commonplace. Therefore, addressing antimicrobial resistance by developing antibiotic-sparing therapeutics is an urgent global health concern. Urinary tract infections (UTI) drive over 15% of all antibiotic prescriptions and directly contribute to the development of AMR bacteria. One potential antibiotic-sparing therapeutic for UTIs is monoclonal antibodies (mAbs), which have been successfully deployed for decades and have a strong history of safety and efficacy. The objective of this proposal is to develop mAbs to two types of UTIs that greatly contribute to global disease burden. The overall hypothesis is that mAbs to bacterial pilus adhesin proteins will block adhesin-ligand interactions and thus prevent bacterial adherence to host tissues. In Aim 1, mAbs will be explored as a treatment for catheter- associated UTI (CAUTI) caused by two pathogens that are frequently multi-drug resistant: Enterococcus faecalis and Acinetobacter baumannii. These bacteria cause CAUTI by using sticky adhesins to bind to fibrinogen deposited on the surface of urinary catheters. mAbs will block this interaction to prevent catheter colonization. In Aim 2, mAbs will be tested for their ability to block bacterial interaction with host tissue. Uropathogenic Escheriscia coli (UPEC) frequently causes highly recurrent UTI (rUTI) in part by establishing reservoirs in the gastrointestinal tract and vagina that serve as a source for UPEC’s continuous reintroduction into the bladder lumen. While the adhesins responsible for gut colonization have been characterized, the adhesin responsible for vaginal colonization is unknown. Based on existing data suggesting a role for the UPEC S pilus in the vagina, the contribution of this pilus to vaginal colonization will first be elucidated. mAbs will then be generated to the S pilus adhesin and tested for their ability to deplete UPEC from the vagina. The long-term goal of the proposed research is to generate mAbs that can treat human urinary tract infections. During the fellowship, the applicant will develop important skills for becoming an independent investigator of infectious diseases. The sponsor of this work, Dr. Scott Hultgren, has vast experience studying urinary tract infection pathogenesis and treatment, and the institutional environment provides supportive, collaborative experts in microbiology and immunology. Washington University School of Medicine has a long history of helping physician-scientists build successful careers. The proposed training plan will facilitate the applicant’s transition into becoming an independent physician-scientist, using research to improve women’s health.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY / ABSTRACT Developmental dysplasia of the hip (DDH) is a life-long disorder that alters joint biomechanics and increases the risk of early osteoarthritis. The most common treatment for DDH is periacetabular osteotomy (PAO), which surgically reorients the abnormally shaped acetabulum to better cover and stabilize the femoral head. After PAO, patient reported outcome measures (PROMs) improve for many, but not all, patients, and PROMs for most patients remain below the levels of their healthy peers. Recent work has shown that aberrant biomechanical variables, including disproportionate joint reaction forces, high acetabular edge loads, and low abductor muscle strength, are related to specific geometric deformities in DDH prior to PAO. These altered biomechanics are also related to worse PROMs. The current project seeks to optimize surgical care for DDH by determining the effects of, and relationships among, PAO-induced changes to hip geometry, biomechanics, and PROMs. Aim 1 will assess how PAO-induced changes in bony geometry alter hip biomechanics. Geometric and biomechanical profiles for patients (N=60) undergoing PAO will be established before surgery using magnetic resonance imaging, motion capture, and musculoskeletal modeling. Changes to patients’ biomechanical profiles resulting from PAO-induced changes to geometry will be determined at 6 and 12 months post-PAO. Aim 2 will establish target ranges for PAO surgical correction based on the sensitivity of biomechanical outcomes to variability in acetabular reorientation. Surgically feasible combinations of changes to patients’ acetabular geometry will be tested using probabilistic analysis, musculoskeletal models, and high-throughput computing to predict the resultant changes to biomechanics. Aim 3 will determine the relationships among PROMs, activity levels, and biomechanics pre-PAO and over the first 12 months post-PAO. Electronic questionnaires and wearable sensors will be used to monitor PROMs and activity changes as patients recover from surgery. From these data, we will demonstrate how hip biomechanics and activity levels differ in patients with improved PROMs compared to those with unresolved symptoms. Results from the project will inform customized surgical plans according to patients’ initial bony geometry and help surgeons and patients focus on modifiable treatment factors that are associated with improved outcomes. Likewise, new information about variables such as post-PAO activity levels and muscle strength will inform rehabilitation to further improve outcomes. Finally, this project will pave the way for long-term longitudinal analyses of PROMs, biomechanics, and joint structure to inform surgical and rehabilitation approaches that predictably mitigate early hip osteoarthritis.

NIH Research Projects · FY 2026 · 2023-02

This application would create a neuroscience training program in St. Louis (NeuroPREP) for recent undergraduate students. The objective of the grant is to provide rigorous and critical training in neuroscience to a cohort of students taking advantage of the strong interest in neuroscience at Washington University, the University of Missouri-St. Louis, St. Louis University and Harris-Stowe State University. By providing fully-funded support for 2 years of independent research and an introduction to the culture of science, this proposal will establish a pathway to graduate training in Neuroscience. NeuroPREP emphasizes sustained training in oral and written science communication, discovery science and outreach experience. Specifically, this proposal will support 7 early-stage trainees for two years each (3 of whom will be supported with Institutional resources). Washington University has long-standing commitments to cutting-edge research, to interdisciplinary education, and to providing modern career development. We seek to be a Program that responds to changes in the research environment by helping our students to pursue important and innovative problems and concepts, to adopt new techniques and to communicate effectively with their peers and the general public.The potential research environment is organized as a Training Faculty of 54 active scientists who study fundamental questions in basic and clinical neuroscience. The 54 faculty represent 16 different Departments in three different Schools at Washington University (Arts & Sciences, Medicine, Engineering). Each faculty mentor directs a well-funded research program and is deeply committed to provide a positive learning experience and welcoming work environment. The curriculum for education will likewise present a broad and deep introduction to molecular, cellular and systems-level approaches to the study of neural function and dysfunction. It will organize dedicated courses, but also offer optional involvement in existing undergraduate and graduate coursework. The Program will recruit and retain talented students through innovative and dedicated coordination with the University and partner schools and be evaluated formally by an Executive Advisory Board of Directors. Major new initiatives aimed at accomplishing these goals include: 1) creation of a local network of educators across many Schools to identify candidates and foster the program for the benefit of the region; 2) the introduction of two interactive courses to bolster neuroscience fundamentals and a sense of community among the students (Critical Thinking in Neuroscience & Professional Skills for a Neuroscientist), 3) involvement of the students in the Society for Neuroscience Brain Bee as part of their training in science communication, and 4) refinement of a near peer-mentoring program that has graduate students working with postbaccs and postbaccs working with high school students. These initiatives will promote our students’ education in neuroscience, and in professional skills. It will also provide them explicit guidance in the process of graduate school applications.

NIH Research Projects · FY 2025 · 2023-02

Project Summary/Abstract The emerging role of inflammation in the pathogenesis of Alzheimer’s disease (AD) is shifting how the field is approaching its treatment, with growing interest in the development of immunomodulatory therapeutics. The appropriate therapeutic window and patient population for such treatment strategies remains to be determined. Importantly, current understanding of inflammation in AD has largely arose from brain tissues studied in isolation. However, it has become clear that peripheral inflammatory responses may influence both AD risk and disease progression. Here, I propose to use whole-body positron emission tomography (PET) of the translocator protein 18kDa (TSPO) and triggering receptor on myeloid cells 1 (TREM1) and complementary immune profiling techniques to non-invasively assess peripheral and central inflammation in Alzheimer’s disease. This project aims to use parallel preclinical (Aim 1) and clinical approaches (Aims 2 & 3) to increase the fundamental understanding of inflammation in disease while actively improving clinical assessment. Our specific aims are (1) to characterize distinct peripheral and central myeloid cell responses and investigate the effects of systemic inflammation on neuroinflammation in the 5XFAD mouse model of AD; (2) to develop a clinically feasible approach to quantify whole-body TSPO-PET uptake in AD patients; and (3) to study whole-body immune signatures associated with disease severity in AD and mild cognitive impairment patients using whole-body TSPO-PET and blood-based immune profiling. The innovation of this work lies in the whole-body approach for the investigation of inflammation in AD, which has yet to be investigated. TREM1-PET is the first tool to specifically image proinflammatory peripheral myeloid cells in vivo. Preclinical investigation using TREM1-PET and clinical application of whole-body TSPO-PET imaging in patients will provide novel insights into the complex neuroimmune interactions involved in AD pathogenesis. The significance of this work is that enhanced understanding of whole-body innate immune responses in AD has the potential to not only improve diagnosis and disease monitoring, but also to develop and screen for effective disease modifying therapeutics. Additionally, these methods can be applied to impact our understanding of inflammation across a broad range of inflammatory and neurological disorders.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY/ABSTRACT The steady erosion of cognitive function is a hallmark of aging and markedly exacerbated in Alzheimer Disease (AD). These deficits have no effective treatments. Depletion of nicotinamide adenine dinucleotide (NAD+) and reduction of NAD+-dependent sirtuin activity in aging and AD have been well documented. Restoring NAD+ or activating the sirtuin SIRT1 has induced abatement of aging symptoms including cognitive decline. However, current NAD+ replenishment strategies are non-specific and the success of SIRT1 activation as an anti-aging therapeutic may require tissue-specific activation and the concomitant restoration of NAD+. We have shown previously that nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme in the major mammalian NAD+ biosynthesis pathway, is contained in extracellular vesicles (EVs) and secreted to plasma. Treatment of aged mice with plasma-derived EVs containing extracellular NAMPT (eNAMPT) extends mouse lifespan and healthspan. Our preliminary data suggest that EV treatment can ameliorate cognitive dysfunction and rescue age-related decreases in hippocampal CA1 synapse counts in 24-month-old mice. EVs are also targeted to the brain and can increase NAD+ in cultured neurons. However, the tissue-specific EV targeting mechanism as well as the implications for EV treatment in the context of AD-related cognitive decline remain unknown. This study will test the hypothesis that EV-contained eNAMPT rescues age-related cognitive dysfunction via activating SIRT1 in key neuronal populations and that leveraging this pathway can lessen AD- associated cognitive decline. In Aim 1, cell culture-derived EVs will be utilized to test the necessity of eNAMPT in the targeting and uptake of EVs both in vitro and in vivo. In Aim 2, the possibility that the cognitive benefits of EV treatment occur through eNAMPT-mediated activation of SIRT1 will be evaluated. This determination will be accomplished via EV treatment with and without EV-contained eNAMPT and with and without hippocampal knockdown of Sirt1 by viral delivery of shRNA. Aim 3 will test the effectiveness of EVs in treating cognitive deficits and pathological progression in a mouse model of AD. Therefore, this study has the potential to elucidate a novel role for eNAMPT in facilitating both the cellular targeting of EVs and the efficacy of EVs in rescuing age- and AD-related cognitive dysfunction. This project may also establish eNAMPT-containing EVs as a viable biologic to treat cognitive deterioration in aging and AD. This fellowship proposal integrates the NAD+ biology and aging expertise of sponsor Dr. Shin Imai with comprehensive training in behavioral neuroscience, AD models, advanced synaptic analysis, EV engineering, and laboratory mentorship and leadership. This training will be further empowered by the intellectually stimulating and highly collaborative environment of Washington University in St. Louis. Thus, this fellowship will constitute ideal preparation for the applicant’s career as the future leader of an academic laboratory.

NIH Research Projects · FY 2025 · 2023-02

Project Summary: Diversity at the molecular level has created the riot of life forms ever to exist on earth. Diversity of thought, perspective, and background among individuals working as part of a team enhances performance. The pursuit of scientific knowledge and excellence then demands the inclusion of students from all backgrounds. This application requests funds to support an Initiative for Maximizing Student Development (IMSD) program within the Division of Biological and Biomedical Sciences (DBBS) at Washington University in St. Louis at the level of ten trainees, equal to our current level of support. The mission of our IMSD program is to increase the matriculation, training, retention, graduation, and career outcomes of outstanding PhD students from groups historically underrepresented in the sciences in order to help change the face of the next generation of scientists and thus increase the power of the STEM workforce in the US. Since its inception in 2013, our IMSD program has created 15 training elements that integrate seamlessly with PhD student training and research, bolstering the academic, professional, and career success of essentially all entering underrepresented (UR) PhD students and often all DBBS PhD students. Concentrated in years one to three, these training activities span our students’ graduate careers and focus on ensuring they surpass defined academic milestones, learn to think critically and to write and speak effectively about research, develop strong student support networks, explore career options, and can engage in community-based educational outreach activities. Since their creation, these activities and our IMSD Program have helped reduce and of late appear to have eliminated the previously persistent achievement gap between URM and well-represented students in terms of qualifying exam success and PhD student retention/completion. Our new IMSD Program will welcome all entering UR DBBS PhD students into it, ensuring that it remains the epicenter of academic, scientific, and career support for UR students in DBBS. We propose to support students for their first two years of graduate school and will preferentially select students who have overcome significant hardship for support. Over its nine-year history, our current IMSD program has increased UR student success and serves as the foundation for our new IMSD program. By focusing on excellence and innovation in graduate education and community-building, our new program will accelerate the tradition of scientific excellence in the US by increasing the presence and success of diverse scientists in our PhD programs and ultimately in the STEM workforce.

NIH Research Projects · FY 2025 · 2023-02

The over-arching goal of the Washington University (WU) StARR Program in Cross-Disciplinary Oncology Clinician Scientist Training is to IDENTIFY, RECRUIT, TRAIN, RETAIN, and ADVANCE outstanding physicians in oncology-focused graduate medical education (GME) training (i.e. “residency”) for careers as clinician-investigators. We have proposed training clinician resident-investigators from multidisciplinary oncology-focused specialties and residency training programs. Long-term objectives: The WU NCI R38 StARR program will successfully identify, recruit, and train oncology-focused clinicians with an interest in becoming clinician-scientists during their residency training. These trainees would then be eligible for subsequent career-development awards, including the companion K38 mechanism. Our program plan includes continued long-term follow-up and engagement of R38 trainees to retain and advance their careers as physician-scientists. Key Elements of Training: An emphasis of our proposal includes multiple levels of career development, including co-mentoring by at least two mentors in different departments to encourage cross-disciplinary training and collaboration. Each resident-investigator will also have a mentoring committee and report regularly to the PI and Advisory Committee. A structured curriculum with specific competencies and milestones will be established to best train individual resident-investigators in programs and research programs that are ideally matched with their skills, interests, and career development. We have received broad support from all levels of research and education at WU, including the Dean’s Office and the participating Departments and Programs. Research Projects: The WU NCI R38 StARR program will support up to four resident-investigators per year from the participating departments. Each resident-investigator will engage in 1-2 years of dedicated, protected research time, free from all clinical responsibilities. They will craft a research plan commensurate with their interests and prior experiences that may involve further training, coursework, workshops, and career development. Specific metrics and plans for follow-up, evaluation, and long-term mentoring and career development are included in our proposal. The WU StARR Program in Cross-Disciplinary Oncology Clinician Scientist Training will leverage the significant institutional, departmental, and programmatic resources at our University to meet the stated goal of the StARR program to recruit and retain outstanding health professionals to foster their careers as clinician-investigators.

NIH Research Projects · FY 2026 · 2023-02

ABSTRACT Parkinson disease (PD) is a progressive neurodegenerative disease characterized by motor, cognitive, and psychiatric manifestations resulting from abnormal protein deposition and neurotransmitter deficits. The variability in clinical presentation and progression in PD likely reflects underlying variability in brain pathology. Although current treatments provide dramatic motor benefit in PD, they fail to fully alleviate gait impairment and non-motor symptoms and may exacerbate cognitive and psychiatric features. These more complex symptoms are linked to the function of large-scale brain networks, which can be measured with resting-state functional connectivity MRI (RSFC). In our past work, we demonstrated that PD participants, as a group, show differences in RSFC relative to healthy controls. However, development of clinical applications requires reliable individual- level biomarkers that capture the widespread neuropathology and respects the clinical heterogeneity of PD, opening the avenue to “personalized medicine” in PD. Recently developed precision-mapping RSFC approaches now permit identification of individual-level differences in brain network organization with high reliability and may provide a non-invasive biomarker for PD. Therefore, we propose to identify individual-level RSFC markers of PD, examine the relationship of these precision RSFC markers with the clinical manifestations and neuropathology of PD, and determine if precision RSFC markers predict cognitive decline and dementia in PD.

NIH Research Projects · FY 2026 · 2023-02

Project Summary: Low levels of physical activity (PA) are common in individuals who use wheelchairs due to physical disability. Wheelchair users (WU) are also at greater risk for obesity and cardiometabolic health-related diseases compared to the general population. Regular PA is widely recognized as being beneficial to the health of persons without a disability and is believed to hold similar benefits for WU. However, WU face barriers to PA, including considerable lack of accessible, community-based facilities, limited access to adapted equipment, lack of knowledge on how to exercise safely, and proper support. Additionally, it is unclear what type of PA intervention is most effective to facilitate WU in achieving current PA recommendations and reversing or preventing cardiometabolic health-related diseases. Thus, evaluating a structured, community-based PA training intervention with education and support is critical and will have significant implications for WU health, future PA guidelines for WU, and the activities of community-based organizations serving WU. The extensive health benefits of aerobic and strength training are well established for the general population, and randomized controlled trials studying the impact of PA for WU suggest programs that provide support and education in accessible, community-based environments are effective for improving outcomes such as strength and pain. However, there is insufficient evidence on whether these interventions result in cardiometabolic health (i.e., VO2peak) improvements in WU. Previous evidence demonstrates these interventions may be inadequate to reach the recommended frequency, intensity, and duration of PA. Preliminary data suggest guiding PA intensity may be essential to improving cardiometabolic health outcomes in WU. The current study proposes a hybrid I randomized controlled trial to test the cardiometabolic effects of a tailored intensity-controlled physical activity training (I PAT) intervention compared to education and access to a community-based accessible gym (EA). The following specific aims will be tested in WU randomized to either the IPAT or the EA intervention groups (N = 54 for both). Specific Aim #1: To compare the effectiveness of the IPAT to EA on cardiorespiratory fitness, vascular function, and body composition. Specific Aim #2: To identify barriers and facilitators to WU engaging in PA at a community-based, accessible gym. Specific Aim #3: To examine the mediators (e.g., self-efficacy) and moderators (e.g., age, race, duration of disability) of the expected intervention effect to understand differences in physiologic response. Moderate-to-vigorous intensity aerobic and strength training will be used to study the effects of interventions delivered in a community-based accessible gym on cardiometabolic related health outcomes such as cardiorespiratory fitness (VO2peak) and vascular function (endothelial function) in WU. Successful completion of the proposed study will generate data to inform future PA guidelines and strengthen the evidence base for safe and effective PA interventions.

NIH Research Projects · FY 2026 · 2023-01

Project Summary/Abstract This work focuses on enterotoxigenic Escherichia coli (ETEC), globally the most common bacterial cause of serious diarrheal illness. Originally identified more than 50 years ago as a cause of severe diarrheal illness in patients with clinical presentations indistinguishable from cholera, ETEC continue to threaten the lives of many children in poor regions of the world where sanitation and clean water remain limited. While deaths from diarrheal illness in low-middle income countries have declined, largely due to deployment of oral rehydration therapy, the tremendous morbidity associated with these illnesses remains completely unchecked. ETEC are closely linked to a condition known as environmental enteropathy or environmental enteric dysfunction (EED) that is complicated by sequelae of malnutrition, stunted growth, and intellectual impairment, robbing poor countries of badly needed human capital. Children with malnutrition are also at tremendously increased risk of dying from infectious diarrhea, pneumonia, and other common infections. In addition, ETEC have been linked to a centuries-old enigmatic disease, known as tropical sprue, that is typically diagnosed in adults with substantial repeated exposure to these pathogens. With EED it shares features of poor nutrient absorption, wasting, and alterations of small intestinal morphology. Much is known about of the molecular pathogenesis of acute diarrheal disease, with heat- labile (LT) and heat- stable (ST) toxins signaling through cAMP and cGMP second messenger pathways, respectively, to alter enterocyte ion channels that promote the net efflux of salt and water into the intestinal lumen. However, the biology underlying enteropathic changes to the small intestine and related sequelae are very poorly understood. Our recent studies indicate that ETEC toxins compromise cellular messages critical to the maintenance, biogenesis and function of small intestinal surfaces responsible for nutrient absorption. The current project will therefore address the following questions: · “What is the impact of ETEC and its individual toxins on the absorptive architecture of small intestine?” · “How does repeated ETEC infection affect the overall expression of genes that direct formation of surfaces needed for nutrient absorption?” · “Can we mitigate these effects by toxin-neutralizing vaccination?” Addressing these fundamental questions will fill important gaps in our understanding of the sequelae to ETEC infections, elucidate important features of the molecular pathogenesis of disease, and inform strategies to prevent illnesses that threaten millions of disadvantaged children worldwide.

NIH Research Projects · FY 2026 · 2023-01

Abstract Expanding participation and opportunity within the Pediatric Neurosciences workforce is essential for improving health and accelerating research toward innovative treatments for pediatric neurological disorders. Children in communities with decreased access to healthcare and advanced scientific educational resources, such as inner-city neighborhoods, are impacted by cognitive and neurological conditions at higher rates, yet they often encounter fewer options for engagement with neuroscience research. Agencies such as NIH/NINDS have taken steps to address these workforce limitations by supporting resource development, research education initiatives, and partnerships to broaden early exposure to neuroscience careers. This proposal will establish the St. Louis Summer Program Immersion in Neurosciences (SLSIP), providing a rigorous research education experience for high school students who would otherwise lack access to research training programs. The SLSIP R25 program is innovative and directly aligned with NINDS priorities for expanding workforce representation, fostering scientific creativity, and improving child health outcomes in the St. Louis region and beyond. The program will develop and support: 1) an expanded candidate pool; 2) transparent and merit-based recruitment; 3) extensive outreach and networking; and 4) formative relationships. Washington University's substantial infrastructure and collaborative environment make it an ideal setting for this unique training opportunity. By advancing the goals of PAR-21-168, SLSIP will strengthen the collective mission of NINDS, WUSM, and the St. Louis community to improve access to research education in Pediatric Neurosciences, ultimately improving health outcomes for all children.

NIH Research Projects · FY 2026 · 2023-01

The goal of the NRCDP is to recruit, identify, and support promising neurosurgeons at the beginning of their careers, when they are most vulnerable, so that they can successfully transition into independent neurosurgeon-scientists. The first 10 years of the program were tremendously successful. During the next five years, we aim to build on this success by recruiting and retaining a broad group of neurosurgeon-scientists across all subspecialties, creating a “Pipeline Program” for recruiting the most talented group of K12 applicants, expanding the “Training Program” to accelerate timing to first independent NIH award, and establishing an NRCDP leadership structure that embraces continuous renewal and evolution. This renewal application would provide funding to administer the Neurosurgeon Research Career Development Program (NRCDP) for another five years. This program is the basis of a continued national effort to support, train, and mentor junior neurosurgical faculty members at appropriate institutions nationwide. The proposal addresses the rationale and ongoing need for the NRCDP and includes several proposed innovations to augment and expand the scope of the program, the leadership structure, and the rationale for the selection of the National Advisory Committee (NAC). It also details a well-developed and robust system for the recruitment and selection of scholar-applicants based on their potential, their mentor’s track-record, and the support of their Chairs. In addition, there is a description of the Annual Retreat, which includes applicant interviews, symposia on reproducibility and rigor, interactive grant-preparation sessions, and NIH style “study section”, where the NAC reviews applications and selects Scholars. Eligible candidates are newly appointed neurosurgical faculty within one year of completing residency or fellowship. Successful applicants are called Scholars. Qualified applicants not selected for funding are called Emerging Investigators and are also an important part of the program. The primary goal of the program is to support Scholars along the path to scientific independence. Hence, an important metric is success in securing subsequent independent funding from the NIH, or other federal agencies. In the 10 years the program has been in place, we have reviewed applications from 138 individuals, at 75 different institutions, located in 36 states. In the most recent analysis, we found that the success rate for Scholars obtaining subsequent NIH funding increased significantly year by year – approaching 85% by nine years. These benefits did not only accrue to Scholars, but also to Emerging Investigators. Their success rate in obtaining subsequent independent NIH funding also grew over time, though at a slower rate, and approached 60% by nine years, due to the development of a parallel path for continued mentoring in the Academy Emerging Investigator Program (EIP). In this competing renewal, we have identified four important priorities: Goal One: To promote efforts to recruit and retain a broad group of neurosurgeon-scientists across all subspecialties. Goal Two: To create a “Pipeline Program” to recruit the most talented group of K12 applicants. Goal Three: To expand the “Training Program” to accelerate timing to first independent NIH award. Goal Four: Establish an NRCDP leadership structure that embraces continuous renewal and evolution. Achievement of these goals will lead to a cadre of independent neurosurgeon scientists that perform ethical, rigorous, and high-impact research into the pathogenesis and treatment for many of most disabling disorders affecting the nervous system.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Sarm1 (sterile α and TIR motif-containing protein-1) is a NADase enzyme that is highly expressed in the nervous system. The Sarm1 enzyme serves as a metabolic biosensor and is activated in response to injury, inflammation, and oxidative stress. Based on a screen of thousands of candidates in 2013, Sarm1 was identified as the central executioner of nerve axon degeneration. Since this time, Sarm1 has also been shown to modulate nerve function through regulation of MAPK signaling and metabolite turnover. In response to this pivotal discovery, Sarm1 inhibitors are now being developed for clinical management of neurodegenerative disease. Specific to bone - nerve damage, dysfunction and clinical neuropathy have all been related to fracture risk and impaired bone health in diverse conditions including spinal cord injury, anorexia, chemotherapy, multiple sclerosis and diabetes. However, the molecular mechanisms underlying these relationships remain unclear, limiting our options for therapeutic intervention. Recently, we discovered that knockout of Sarm1 prevents bone fragility in diabetic mice. Our central hypothesis is that neural Sarm1 activation restricts bone formation, leading to decreased bone mass and strength. Conversely, we hypothesize that targeted Sarm1 inhibition can be used to simultaneously promote bone and nerve health in states of chronic Sarm1 activation, such as diabetes. To test this hypothesis, we will pursue two specific aims. First, we will isolate the role of Sarm1-dependent neuropathy in the progression of skeletal disease. Second, we will target the function of Sarm1 to restore bone health in vivo. When complete, this work will define the mechanisms linking nerve damage to impaired bone health through Sarm1. We will also determine if Sarm1 inhibition is a strategy that can be used to support bone formation in settings of skeletal disease. Our long-term goal is to promote lifelong health and healthy aging by developing strategies to prevent or to reverse nerve and bone damage across diverse disease states, beginning in childhood and adolescence and continuing throughout the lifespan.

NIH Research Projects · FY 2026 · 2023-01

Project Summary Among patients with cancer, the risk of anticoagulant-related major bleeding is more than double that of patients without cancer. These bleeds have a fatality rate of 7%. Current guidelines recommend indefinite anticoagulant therapy in many patients with cancer and venous thromboembolism (VTE) with periodic assessments of bleeding risk. However, no evidence-based models exist to quantify bleeding risk in patients with cancer taking anticoagulants. In addition, recent trials have demonstrated effectiveness of direct oral anticoagulants as primary prophylaxis for prevention of initial VTE in patients with cancer. Thus, there is a strong need to balance the competing risks of VTE and anticoagulant-related bleeding in patients with cancer. While there are models that quantify bleeding risk in non-cancer populations (i.e. atrial fibrillation, non- cancer associated VTE) taking anticoagulants, they have not been validated in patients with cancer. Quantification of bleeding risk in patients with cancer and VTE would allow for an evidence-based approach to preventing and treating VTE by minimizing anticoagulant therapy in patients at high-risk of bleeding and allowing for judicial use in patients at low-risk of bleeding. The overall objective of this study is to develop a prediction model for anticoagulant-related bleeding in patients with cancer and VTE on anticoagulant therapy and to calibrate that model in cancer patients at high-risk of VTE being considered for primary thromboprophylaxis with anticoagulant therapy. We will accomplish our overall objective through the following specific aims: (1) validate eight existing bleeding prediction models in a cohort of 7489 patients with cancer and VTE receiving anticoagulant therapy, (2) refine and validate a cancer-specific prediction model to quantify risk of anticoagulant-associated bleeding in patients with cancer on anticoagulant therapy for VTE, and (3) calibrate the refined bleeding prediction model in a cohort of patients with cancer receiving primary anticoagulant thromboprophylaxis enrolled in two randomized trials (AVERT and CASSINI). These aims will be addressed with an expert research team in the areas of bleeding, VTE (especially in patients with cancer) and development and implementation of risk prediction models. This proposal is innovative as it addresses a current gap in the management of patients with cancer with or at high-risk of VTE by developing a prediction model to quantify the risk of the intervention (anticoagulant therapy) to inform and guide patient care. This research will be significant because it has the potential to decrease anticoagulant-related bleeding and thus bleeding related morbidity and mortality in patients with cancer.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY/ABSTRACT Understanding circuit-level maneuvers that affect brain plasticity will inform the design of targeted interventions after stroke. Experiments outlined in this proposal will determine the contributions of excitatory/inhibitory circuits on brain repair processes after focal ischemia, and how changes in behavioral performance relate to cell-specific changes in connectivity. Stroke causes direct structural damage to local brain circuitry and indirect disruption of global networks resulting in behavioral deficits spanning multiple domains. Stroke recovery is associated with functional brain reorganization, a process involving the formation of new or alternative circuits. Along with behavioral recovery, damaged regions remap to adjacent tissue while patterns of resting-state functional connectivity (RS-FC) within and across resting-state networks gradually renormalize. While local and global changes in functional brain organization are consistently observed during recovery, how these processes relate to the underlying neuronal circuitry supporting recovery of function is unknown. This knowledge gap exists partially because stimulus-evoked and resting-state patterns reflect ensemble activity from many cell types, and patterns of RS-FC can be orchestrated through indirect pathways. Understanding how disconnected inhibitory and excitatory circuits reintegrate into global networks to support recovery requires examination of neural network connectivity structure as it evolves with neuroanatomical markers of circuit repair. While an integrated mechanism relating cellular plasticity with network plasticity has yet to be established, inhibitory circuits have been shown to play a key role. Stroke disrupts the brain’s balance of excitation and inhibition. Restoring this balance through non-invasive brain stimulation techniques can improve recovery. However, treatment efficacy using these methods is extremely varied, partially due to the imprecision and indiscriminate activation or inhibition of all cells near the stimulated site. Parvalbumin interneurons (PV-INs) are the most prevalent of all GABAergic interneurons, play key roles in shaping excitability over long distances, and regulate functional brain rhythms reflected in coherent patterns of RS-FC. Though their role in post-stroke plasticity is unknown, PV-INs are known to mediate several other forms of activity-dependent plasticity, making them compelling candidates for affecting repair processes after stroke. Using optogenetic targeting and wide field optical imaging of cortical calcium dynamics in awake mice, we will establish functional connectomes of excitatory (CamK2a-based) and inhibitory (PV-based) circuits and how they evolve following focal ischemia (Aim 1). We will utilize the well- characterized motor-barrel network in the mouse to directly test the influence of activity in cortical excitatory/inhibitory nodes exhibiting strong (Aim 2) or weak (Aim 3) inter- or intra-hemispheric connectivity with perilesional tissue, and how these manipulations affect neuroanatomical markers of circuit repair. At the conclusion of this grant, we will determine the contributions of CamK2a/PV circuits on post-stroke recovery, and further understand the components of connectivity restoration required for more complete behavioral recovery.

NIH Research Projects · FY 2026 · 2023-01

Anxiety disorders are the most common form of pediatric psychiatric illness, affecting up to 30% and severely impairing up to 20% of all youth prior to age 18. Unfortunately, up to 50% of children remain symptomatic even with best available treatments, making anxiety disorders a major public health problem. To devise new treatments for anxiety disorders, more research is needed into underlying brain mechanisms. Research studying mechanisms suggests that anxiety disorders are linked to alterations in attention, including increased attention to threatening stimuli (e.g., feared objects or sounds). However, more recent research suggests these attention alterations may be due to a larger problem in which attention is increased to all stimuli (e.g., any loud noise, flash of light, etc.). This alteration in attention seems to be the opposite of ADHD, the prototypical disorder of attention in childhood and also a common childhood psychiatric disorder. In ADHD, children appear to have a generalized decrease in attention to suddenly appearing stimuli (e.g., children with ADHD might not hear their name called). Intriguingly, although anxiety and ADHD appear to demonstrate opposing attention- related problems, these disorders occur in the same child more often than expected by chance. Understanding the brain mechanisms underlying attention alterations in pediatric anxiety and ADHD is critical in designing new treatments that attempt to target or ‘correct’ problems in these brain mechanisms. In this study, we test the hypothesis that attention-related brain circuitry is overactive in anxiety disorders such that bright objects and errors elicit increased activity compared to children with no disorder. We further predict this circuitry is underactive in ADHD such that bright objects and errors elicit diminished activity increases. We hypothesize that children with both anxiety and ADHD have an intermediate level of activity (similar to children with no disorder), though less is known in this area. Finally, we predict that peer observation (a mild threat/stressor) further exacerbates the overly active attention-related brain circuitry in children with anxiety. To test these hypotheses, we use cutting-edge neuroscience tools to provide a nuanced characterization of attention-related brain circuitry in N=300 children ages 10-12 years with anxiety disorders (n=75), ADHD (n=75), both anxiety and ADHD (n=75), and no psychiatric disorder (n=75). Children play a computer game during which we measure how suddenly appearing objects capture their attention. Brain activity is measured using functional MRI, pinpointing the specific brain locations that are disrupted; and electroencephalography (EEG), providing precise timing information. We characterize attention-related brain circuitry when children are being observed by a peer versus without this stressor. This study will provide a comprehensive description of circuit-level mechanisms of altered attention in pediatric anxiety, ADHD, and co-morbid conditions, providing mechanistic targets for novel treatment design. Results could ultimately have major public health impact, by assisting in the design of new treatments for two of the most prevalent and impairing childhood psychiatric disorders.

NIH Research Projects · FY 2026 · 2023-01

Abstract Chronic pain is debilitating disease that affects more Americans than cancer, heart disease, and diabetes combined. Despite this significant public health problem, effective treatments are scarce and commonly prescribed opioids possess significant abuse liabilities. One possible reason for this poor translational success is that we still lack a detailed understanding of how these circuits are connected to process sensory information and their plasticity mechanisms. In our preliminary experiments we have identified important roles for trans-synaptic adhesion molecules in regulating somatosensory synapse function in the spinal cord. Here we will determine the trans- synaptic molecules that influence somatosensory synapse formation, understand how presynaptic adhesion molecules in somatosensory neurons instruct the formation and function of native synapses in the spinal cord, and establish their role in coordinating nociceptive circuit assembly to regulate pain behaviors. This proposal will use a combination of in vitro synapse induction assays, conditional gene knockout and rescue approaches, peripheral viral circuit tracing, optogenetic slice recordings, and somatosensory phenotyping to understand how trans- synaptic adhesion molecules regulate somatosensory circuit assembly and function.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY The intestinal epithelium perpetually self-renews and differentiates into specialized progenies. This process is often viewed as a hard-wired genetic program under the control of host-derived factors. The organizing principal of my proposal is that specification, differentiation and functional specialization of these epithelial lineages are modulated by members of the gut microbiota and their metabolic products. Our groups' studies of healthy and undernourished Bangladeshi children revealed that Prevotella copri is a key species whose abundance in the developing microbiota is positively associated with ponderal growth. My current work in the Gordon lab has established a gnotobiotic mouse model of mother-to-infant transmission of defined collections of human gut bacterial strains that represent different stages in the postnatal assembly of gut microbiota. I performed single-nucleus RNA-seq to test the effects of including or excluding P copri from these defined communities. A prominent epithelial response to the presence of P. copri is enhanced enterocytic fatty acid oxidation. Pseudotime analysis of snRNA-seq datasets disclosed that expression of fatty acid oxidation genes was spatially regulated by P. copri as enterocytes differentiate/migrate from crypts up villi and that nuclear receptor PPAR appears to be a top candidate regulator of these genes. I hypothesize that i) exposure to the P. copri-containing community broadly alters the epigenetic landscape as well as chromatin accessibility to transcription factor binding as enterocytes execute their differentiation program up the villus, and (ii) PPAR functions as a key transcription factor that mediates the effects of P. copri on fatty acid oxidation with additional regulatory inputs from other transcription factors. I propose two aims to test these hypotheses. In Aim 1, I will characterize, at single-cell resolution, changes in the epigenetic landscape and the transcription factors that comprise the signaling pathways that P. copri-containing community uses to regulate enterocyte differentiation/function along the length of the intestine. In Aim 2, I will characterize how the P. copri-containing defined bacterial community modulates PPAR signaling to control enterocytic fatty acid oxidation along the crypt-to-villus axis, by using a combination of liquid-chromatography mass spectrometry, 2-dimensional organoid culture system, and gnotobiotic mouse models. This research will provide (i) mechanistic insights on how gut microbiota modulates host signaling to control epithelial lineage development, metabolic function, and functional specialization, and (ii) novel metabolites that have therapeutic potential to benefit intestinal physiology and function.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY/ABSTRACT – The incidence of gastric and breast cancers is increasing rapidly, rating fourth and fifth leading causes of cancer mortality worldwide. In patients clinically classified as HER2-positive (ERBB2 amplification and/or 2+/3+ protein overexpression by immunohistochemistry), antibody-drug conjugates (ADC) prolong progression-free and overall survival. However, these therapies have low activity in HER2-low cancer cells, and not all HER2-positive tumors benefit, or even those who initially respond inevitably develop resistance over time. Guided by preclinical data we obtained in HER2 heterogeneous patient-derived xenografts demonstrating that an increase in HER-ADC endocytosis enhances therapeutic efficacy, we developed approaches of antibody delivery that result in a 15.5-fold increase of ADC internalization in cancer cells. Our novel approach uses pertuzumab and trastuzumab-drug antibodies that click at the surface of cancer cells upon binding distinct HER2 domains to increase the number of HER2-ADC complexes and further enhance the rate of HER2-ADC endocytosis. Antibody-PET imaging studies that we have generated demonstrate that our approach increases the uptake of 89Zr-ADC in heterogeneous tumors containing HER2-high and HER2-low cancer cells. Here, we will optimize clickable pairs of two epitope-distinct antibody-ADC biomolecules to enhance tumor targeting and drug delivery in resistant models of breast and gastric cancer. In addition to test the potential of our approach in cancer cell lines and organoids, we will perform randomized imaging and therapeutic studies in patient-derived breast and gastric xenografts representing three tumor populations: ADC-eligible tumors of HER2 heterogeneity, ADC-ineligible tumors, and ADC-resistant tumors. We will determine the molecular imaging (89Zr-Antibody PET), safety, pharmacokinetic profile, and therapeutic efficacy of ADC alone (no-click) versus ADC plus pertuzumab conjugated with clicking pairs (click). These randomized preclinical studies will allow us to identify molecular features that confer drug sensitivity or resistance to this promising investigational approach. Aim 1 will optimize pertuzumab/ADC clicking pairs with improved tumor uptake and drug delivery when compared with ADC monotherapies and Aim 2 will validate the use of antibody clicking pairs as a new therapeutic approach. The two aims will provide important new preclinical data on the use of antibody clicking pairs to enhance drug delivery, which could provide an excellent foundation for many future investigations, including the clinical translation of using clicking pairs to enhance drug delivery and the potential broader application to other membrane receptors and heterogeneous tumors. The long-term translational objectives of the studies proposed are to establish a foundation for a clinical trial using antibody clicking to prevent or delay drug resistance in patients with heterogeneous breast and gastric cancers.

NIH Research Projects · FY 2025 · 2023-01

PROJECT SUMMARY/ABSTRACT Alzheimer disease (AD) affects approximately 6 million people in the United States and currently has no clearly-effective disease modifying therapies. AD is pathologically defined by the accumulation of amyloid β (Aβ) plaques and tau neurofibrillary tangles in the brain and it is known that alterations in the activities of microglia and other non-neuronal cell types play important roles in shaping the disease course. Much of the polygenic risk for AD is derived from variants in genes expressed by microglia, specifically those involved in endolysosomal and lipid processing pathways, and microglia containing abundant lipid droplets (LD-MG) have been observed in post-mortem human AD brains. As previous interventions targeting Aβ have not yet been successful in clinical trials, the concept of modulating microglial lipid metabolism is a novel and exciting avenue being explored in preclinical studies, however we need to know more about how lipid metabolism governs microglial functional states. In mouse models of amyloidosis, microglia transition from a homeostatic to a disease-associated (DAM) transcriptional state that represents a protective, phagocytic, and plaque- compacting phenotype. While microglial activity may be beneficial in the early amyloid phase of AD, pharmacological or genetic inhibition of microglia has been shown to be protective in mouse models of tauopathy. LD-MG are observed in actively degenerating brain regions in a tauopathy mouse model, but the functions of these cells in the disease process are not currently known. To begin addressing this question, we will use FACS coupled with scRNAseq to characterize any transcriptional differences between LDhigh vs LDlow microglia isolated from 9.5 month old tauopathy mice. Given recent evidence suggesting that the LD-MG observed in aged mice have pro-inflammatory, hypo-phagocytic phenotypes, we hypothesize that the LD-MG in our tauopathy model will have similar pathway alterations and could thus be partially responsible for microglial contributions to neurodegeneration. To assess the relevance of these findings to humans, we will stain postmortem human AD brain samples for any promising mouse LD-MG markers. To elucidate the functional roles of LD-MG in the progression of tauopathy, we will utilize a new mouse model that allows for the inducible, microglial-specific knockout of the diacylglycerol acyltransferase (DGAT) enzymes, which have been demonstrated to be required for LD biogenesis in multiple biological contexts. A substantial amount of literature supports a role for lipid droplets in sequestering potentially toxic lipids and we hypothesize that crippling the ability of MG to form LDs via DGAT KO will accelerate tauopathy progression, which we will assess using a combination of immunohistochemical, scRNAseq, and lipidomic analyses. Our studies will characterize an understudied subset of microglia, increase our knowledge of the diversity of myeloid functional states, and evaluate the potential of modulating microglial lipid metabolism for the treatment of Alzheimer disease.

NIH Research Projects · FY 2026 · 2022-12

Birth defects, which occur in 3 - 5% of US-born infants per year, are a leading cause of childhood mortality and repeated hospitalization and are a large burden to families and society. Birth defects typically result from rare genetic changes, but determining which gene-variant causes a phenotype and disease remains challenging, despite significant advances in next-generation sequencing and analysis. Whereas ~4,000 human genes have been linked to monogenic, rare diseases, it has been estimated that 6,000-13,000 additional rare disease genes remain to be identified, many of which are likely causal for birth defects; this underscores a need for effective strategies to assess the functional effects of associated variants. In this application, we propose a multi-organism approach to connect patient presentation with genes not previously associated with disease (genes of uncertain significance, GUS). Focusing on intellectual and developmental disorders and structural birth defects, we will first screen patient birth-defect associated GUS leveraging experimental tractability of C. elegans, Drosophila, or zebrafish, followed by assessment of patient- related phenotypes in vertebrate organisms (zebrafish, mouse, or established human embryonic stem cell lines (hESCs)) for a subset of these genes. In Aim 1, bioinformatic analysis of family based clinical exome sequence will identify high probability candidate gene-variants that may be causal for the patient’s symptom(s) for further study. In Aim 2, nominated candidate gene-variants will be screened in worm, fly or fish to obtain in vivo functional data in support of variant causality. Functional information will help determine whether the gene-variant is damaging, if the gene is required in a specific tissue, and whether the observed genetic mechanism (e.g., hypomorph, dominant negative, etc.) is consistent with patient genetics. Additionally, these experiments may illuminate the molecular or cell biological mechanism disrupted by the gene-variant (e.g., disruption of cytoskeleton, etc.). We will take advantage of the strengths of the different model organisms—CRISPR editing for C. elegans, tissue-specific RNAi in Drosophila, and mRNA and CRISPR embryo injections for zebrafish—in the genetic screening approach. A total of 84 candidate birth defect GUS will be screened in Aim 2 over the grant period. Often, the phenotypic effects of the corresponding disruption of the orthologous gene in worm or fly are not obviously related to the phenotype observed in humans. Therefore, in Aim 3, for a subset of genes (23) identified from the screen as likely disease-causing in Aim 2, we will examine phenotypes in zebrafish, mouse, or hESC systems, to advance our understanding of disease phenotype and progression that is not possible in simple model organisms or with the patient. These studies will leverage our experience in disease gene modeling and our work with clinical collaborators to understand the phenotypic, genetic, molecular, and cell biological basis of each patient’s disease. This innovative multi-organism experimental platform will significantly accelerate identification of birth defect-causing genes and will open avenues to diagnosis, prevention, and therapies.

NIH Research Projects · FY 2026 · 2022-12

PROJECT ABSTRACT This proposal aims to collect data using PET and MRI in humans to measure metabolic correlates of task-evoked neural activity. Our approach is unique in simultaneously measuring the metabolic rates of oxygen (CMRO2) and glucose (CMRGlc), thereby permitting the calculation of aerobic glycolysis (AG), which reflects the rate of excess glycolysis beyond that required for oxidative metabolism. Prior work has shown that the BOLD signal, driven by a change in cerebral blood flow (CBF) that exceeds changes in CMRO2, decreases in particular “task-negative” regions of the brain during task-performance, making up the now so-called default mode network (DMN). It is, however, presently unknown whether AG also decreases or increases in these same regions during a task. Moreover, since Alzheimer’s disease (AD) pathology specifically affects the DMN, it is possible that task-evoked changes in AG are also affected by AD. Here we will collect task-evoked metabolic and BOLD fMRI data in both young and older adults, including adults with and without pathologic evidence of AD. Our primary approach is to determine how task-evoked glycolysis relates to the BOLD fMRI signal deactivations and activations, and to determine whether aging or AD affect these relationships. In Aim 1, we will determine the metabolic correlates of BOLD deactivations during task performance in young adults. We will perform FDG-PET to quantitate regional CMRGlc, as well as MRI measurements of CBF and CMRO2, thus permitting calculation of AG. We will make these measurements alongside BOLD fMRI during rest and during two different tasks. We will then determine whether AG increases or decreases in the default mode network. In Aim 2, we will determine whether task- evoked changes in brain metabolism differ among young adults, older adults, and older adults with AD. We will perform the same studies as in Aim 1, but in older healthy adults (i.e., cognitively unimpaired, amyloid-biomarker negative, without other evidence of neurological disease) and in asymptomatic older adults with pathologic evidence of AD (i.e., amyloid-biomarker positive). We will also choose one of the tasks to further perform in a group of adults with mild AD dementia. From these data, we will determine how healthy aging and AD influence task-evoked brain activity and metabolism. We expect these data to be highly informative, including in the interpretation of numerous other ongoing studies utilizing functional neuroimaging to study how aging and AD influence brain physiology.

NIH Research Projects · FY 2026 · 2022-12

Project Summary Migraine is one of the most common chronic pain conditions worldwide, but prevalence differs based on sex and age, with prevalence of migraine being higher in females. In addition to female sex, family history of migraine represents a further risk factor for migraine. However, the factors which contribute to or protect from migraine onset remain uncharacterized, and it is not yet possible to predict whether an individual will experience migraine onset, even among individuals with both risk factors. Thus, this study aims to investigate: Aim 1 – identify the psychophysical and neural factors which can predict migraine onset in adolescent girls; Aim 2a – determine the hormonal, psychophysical, and neural changes associated with migraine onset; and Aim 2b – characterize the temporal relationships between hormonal, psychophysical, and neural changes preceding vs. following migraine onset. Preliminary data support the use of psychophysical and neural factors as predictors of migraine onset. In addition, changes in pain sensitivity and in the functional connectivity of the amygdala are observed in patients with migraine and following changes in headache frequency, and thus support the use of these assays for the investigation of longitudinal changes related to migraine onset. Study participants will be healthy girls (age 10–13) with either a family history of migraine (Fam-His, N = 160) or with no family history of migraine (No-Fam-His, N = 40). In this study, we will perform assessments of psychophysical (pressure pain thresholds [PPT] and conditioned pain modulation [CPM]), neural (functional connectivity [FC] of the amygdala), and hormonal (testosterone levels) factors at baseline, and at 1- and 2-year follow-up time points. For all study visits, participants will also meet with a pediatric headache/pain specialist to determine whether or not they meet the diagnosis criteria for migraine (participants meeting diagnosis criteria at baseline will be excluded from the study). We hypothesize that a pre-existing pronociceptive psychophysical (lower PPT and lower inhibitory capabilities) and neural (greater FC between the right- amygdala and posterior cingulate cortex) profile at baseline will predispose Fam-His girls to migraine onset during adolescence and can be used to predict this onset. We expect that after 2 years, Fam-His girls diagnosed with migraine will have distinct psychophysical, neural, and hormonal changes as compared to those with no migraine onset and No-Fam-His girls. Moreover, we predict that lower increase in testosterone levels will precede migraine onset, while further transition towards a pronociceptive profile will be observed after migraine onset (reduction in PPT and CPM responses, and increase in amygdala FC). Early identification of individuals most likely to be diagnosed with migraine will facilitate the development of a precision medicine approach, allowing for the implementation of early preventive strategies, thereby reducing patient burden and healthcare system costs. In addition, identifying the sensory, neural, and hormonal mechanisms associated with migraine onset will be foundational to the development of novel migraine treatments.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY/ABSTRACT Generating high quality oocytes is critical for a woman's fertility and health, and for healthy offspring. Nuclear envelope membrane protein 1 (Nemp1) is a transmembrane protein of the inner nuclear envelope that we found is needed for fertility in a wide range of organisms. Loss of Nemp1 in mice leads to near-sterility of females, associated with loss of the oocytes that compose the primordial reserve. The remaining oocytes have poor developmental competence, with defects in chromosome segregation, chromatin compaction and completion of meiosis. GWAS studies show that variants in NEMP1 are associated with early menopause, suggesting the role of Nemp1 in fertility is conserved to humans. Our long term goal is to understand how Nemp1 functions at the nuclear envelope to promote oocyte quality and human fertility. In this grant we will use mouse models to determine why loss of Nemp1 leads to reduced ovarian reserve and poor developmental potential. We will determine when and how the ovarian reserve is lost, and define pathways involved in oocyte loss. We will conduct gain of function and loss-of-function studies, to determine which cells require Nemp1 in the mouse ovary. To identify Nemp1-interacting proteins, we will take advantage of novel mouse lines we have generated, which allow us to conduct affinity-purification coupled mass spectrometry from resting and growing oocytes. Our preliminary studies have revealed that Nemp1 accumulates into extremely large, regular foci at the nuclear envelope, uncovering a novel nuclear envelope structure in growing oocytes. Proteomic analysis will reveal other proteins that localize to these foci (which we term NECs). Super-resolution microscopy and electron microscopy will define the structure of Nemp1 clusters and the adjacent nuclear envelope. FRAP analysis will clarify the stability of the Nemp1 clusters, and associated proteins. Our preliminary data indicate that chromatin compaction is disrupted in Nemp1 mutant oocytes, and that transcription is deregulated. We will use RNAseq to define change in gene expression in Nemp1KO oocytes. We will use ChIP-seq to define changes in histone modifications in mutant oocytes, and determine alterations in 3D chromatin architecture using Hi-C. Our preliminary data indicate that a closely related gene, Nemp2, is upregulated in Nemp1KO oocytes, so we will explore the contribution of Nemp2 to Nemp1 mutant phenotypes. Our preliminary data indicate that the nucleoplasmic tail of Nemp1 can interact with chromatin in cultured cells: we will clarify the regions of Nemp1-chromatin interactions in oocytes, using transgenic DamID approaches. Together these studies will illuminate the function of the nuclear envelope, and the role of Nemp1 in supporting the creation of healthy oocytes.