University Of Minnesota

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Over 75% of people with Parkinson's disease (PD) have significant sleep-wake disturbances that are major contributors to decreased quality of life and can be more disabling and resistant to treatment than the motor symptoms of PD. Currently, the mechanisms contributing to disordered sleep in people with PD are poorly understood and there is a critical need for therapeutic inventions to improve sleep quality. Deep brain stimulation (DBS) has been shown to improve sleep in PD however effects are highly variable across patients. A better understanding of the neuronal mechanisms underlying sleep dysfunction in PD, how DBS affects sleep quality, and the neurophysiological changes and patterns of pathway activation with DBS that underlie these changes would provide the rationale for development of circuit-based DBS approaches to the treatment of sleep disorders in PD. The goal of this proposal is to: (1) characterize the changes in oscillatory activity and connectivity in the basal ganglia-thalamocortical network during disturbances in sleep in PD patients; (2) examine the relative effects of DBS in the STN or GPi on these changes; (3) identify the neural pathways that are preferentially activated (or avoided) in patients with improved or impaired sleep after STN or GPi DBS. We will leverage the well-established infrastructure at the University of Minnesota to externalize DBS leads and perform electrophysiology recordings and stimulation studies in PD patients prior to pulse generator placement (Specific Aims 1 and 2). We will also use high-resolution 7 Tesla (T) MRI, diffusion tractography, and subject-specific computational biophysical modeling to associate pathway activation patterns with quantitative and qualitative measures of sleep outcomes in the year following DBS surgery (Specific Aim 3). This project will increase our understanding of the role of BG-cortical activity patterns on sleep and provide new insights into the mechanisms by which DBS impacts sleep. It will inform the development of more effective stimulation strategies to normalize sleep activity that utilize physiological biomarkers and closed-loop control paradigms tailored to individual patient's sleep-wake cycle. These data will provide the basis to target specific pathways with DBS to optimize sleep-related outcomes in PD.

NIH Research Projects · FY 2026 · 2023-04

Abstract When we learn a complex behavior the nervous system must continuously drive new actions, compare predictions for the actions against outcomes, and strengthen or weaken the connections between neurons (synapses) in order to improve future actions. However, within the multilayer brain networks that control behavior, the behavioral impact of modifying a synapse depends upon many downstream connections. Thus, learning requires the brain solve a ‘credit assignment’ problem: information about which synaptic modifications should be made is distributed across the network, yet must somehow be leveraged by local processes to guide change at individual synapses. A major gap in our ability to relate behavioral events to synaptic change is the current lack of knowledge of these local processes that guide synaptic changes at individual neurons. Recent theories of learning suggest that spikes generated in the apical dendrites of cortical neurons may play a key role in solving this credit assignment problem. The experiments in this proposal will test the hypothesis that the apical dendrites of neurons in the pre-motor cortex integrate multiple learning-instructive feedback sources, and – under appropriate conditions – generate dendritic spikes that rapidly reconfigure the connectivity and function of neurons. In these experiments we will use advanced optical techniques to monitor and manipulate activity in the dendrites of a subset of neurons in the frontal cortex that have a well-delineated role in action planning. A key prediction of our hypothesis is that the activity of the apical dendrites reflects local credit-related calculations and that this activity is distinct from the activity transmitted to other neurons by action potential generation near the cell body. We will test this using longitudinal two-photon calcium imaging of cortical neurons during learning to determine how the behavioral selectivity of dendrites and cell bodies change with changing behavior. In order to identify the contribution of dendritic spikes to learning, we will also use optogenetics to selectively suppress activity in the apical dendrites during learning. Computational models also predict that dendritic spikes are generated by a mismatch between outcome information arriving from long-range feedback projections and local inhibition that predicts this feedback. To test this, we will combine synaptic glutamate imaging and optogenetics to map the selectivity and anatomical identity of feedback projections to the apical dendrites, and calcium imaging to determine the selectivity of local inhibitory neurons that target the apical dendrites. Together, these studies will provide critical new insights into the circuit mechanisms governing cortical plasticity and credit assignment. In doing so, they will provide a key framework for connecting complex learning with modifications at the individual synapse level, and will build bridges between machine learning algorithms and models of biological neural networks.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Our long-term goal is to understand how dynamic regulation of signal transduction systems control cellular stress responses. The focus of this proposal is on identifying the mechanisms by which dynamic expression of the transcription factors p53 and MYC coordinately regulate apoptosis and senescence in response to genotoxic stress. Proper regulation of p53 and MYC are of undeniable importance in human health, as their mutation predisposes human cells to cancer. While the regulation and functions of p53 and MYC have been extensively studied, exactly how they generate variable cell fate outcomes in individual cells of a population responding to the same stress remains poorly understood. Our recent studies have shown that the dynamics of p53, the temporal pattern of p53 accumulation and degradation, serves an integral function for controlling MYC levels and cell fate responses to DNA damage. We have shown p53 dynamics to be highly variable between individual cells, but it remains to be determined how such variability contributes to heterogeneous responses to DNA damaging agents, which is critical for understanding tumor cell heterogeneity and evasion of therapies. To answer this question, we will combine time-lapse fluorescence microscopy to quantify p53 and MYC dynamics with quantitative analysis of key transcriptional targets at the single cell level to determine the temporal regulation of the triggering of apoptosis and senescence in response to DNA damage. We will apply this analysis to three conditions: 1.) non-transformed cells expressing normal p53 and MYC, 2.) transformed cells in which MYC expression is elevated over a range of concentrations, and 3.) transformed cells expressing a p53 gain-of-function hotspot mutation. This work will show how p53 and MYC dynamics control initiation of terminal cell fates in physiological and pathological conditions, and it will serve as the basis for approaches to reduce heterogeneous responses to DNA damaging compounds. These results will provide novel insight into the basic functioning of one of the most important stress response pathways in human cells, and are likely to inform innovative therapeutic strategies based on improved timing of the delivery of therapies. More broadly, this study is likely to provide general insights into the growing list of other important signaling pathways that use dynamic regulation.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT A healthful diet during early childhood is important for healthy growth and development and contributes to the prevention of chronic diseases. Parents influence children’s dietary intake through their use of food parenting practices. Research to date shows positive associations of structure- and autonomy support- food parenting practices with healthful dietary intake and eating behaviors in children, whereas coercive controlling and indulgent practices have been associated with unhealthful dietary intake and the development of maladaptive eating behaviors over time. While research has historically evaluated parents’ “usual” approach to feeding children via questionnaires, recent evidence reveals important within- and between-day variation in use of food parenting practices across time and contexts. Parents have identified a range of momentary factors (e.g., activities, limited time, stress) in everyday family life that alter their usual approach. Parents described shifts from the use of structure- and autonomy support- feeding practices to more indulgent and controlling practices in the face of external challenges. We have recently obtained quantitative evidence of these within- and between-day shifts in food parenting practices through the use of Ecological Momentary Assessment (EMA); EMA uses short surveys delivered to hand-held devices in real time throughout the day to capture dynamic changes in behaviors across time and context. For instance, we have observed that parental stress early in the day is associated with greater use of controlling feeding practices later in the day. Our goal is to build on and extend the evidence-base of food parenting approaches for preventing poor dietary intake among children. We argue that parents’ approach to feeding varies across time and context and that a deepened understanding of the variability in food parenting practices and associated outcomes is necessary to design interventions to help parents maintain consistent use of supportive practices despite challenging circumstances. We propose to comprehensively investigate the impact of within- and between-day fluctuations in food parenting practices on children’s dietary intake overtime using a longitudinal study conducted with a sample (n=240) of racially/ ethnically- and socioeconomically- diverse parent-preschooler dyads. Data will be collected via state-of-the-art measures, including EMA and interview-led 24-hour diet recalls every 6 months for two years. The proposed study represents a significant and necessary next step to inform the development of clinic-based recommendations and public health interventions that account for- and are responsive to- momentary factors found to influence parent’s use of specific food parenting practices.

NIH Research Projects · FY 2026 · 2023-04

Project Summary An outstanding, diverse group of 30 faculty mentors has joined efforts across multiple departments and schools at the University of Minnesota (UMN) to form a new Ph.D. graduate training program entitled, "Inclusive Excellence Training Program in the Systems Biology of Cardiovascular Inflammation." This program fills a major void at our institution by focusing emphasis on the highly clinically relevant problem of inflammation in cardiovascular biology, obesity, diabetes and metabolism with study focus from gene to whole organism. This new predoctoral training proposal addresses a major mission of the NHLBI by focus on physiological systems training of cardiovascular and metabolic inflammation, and this new program will position trainees for diverse successful outcomes in the dynamically changing future workforce. This new training program will also fill a large void in the predoctoral training grant landscape at the UMN. Faculty mentors come from several different colleges and departments across campus, including Integrative Biology and Physiology (IBP), Biomedical Engineering, Biochemistry, Cell Biology and Genetics, Microbiology and Immunology and clinical units in Medicine, including Cardiology and Endocrinology and Metabolism, and Pediatrics. The unique focus and singular effort of this proposal is directed at training the next generation of biomedical scholars in the principles and application of Integrative and Systems Biology. In the purest sense, our focus is distilled into the mechanistic study of human physiology. By any name, whether it be physiology, integrative biology, systems biology, functional genomics, or other, this training grant’s premise is that understanding biological function from cells, to organs, to the whole living organism, represents the leading wave of new knowledge discovery in biomedical science now and in the decades to come. Graduates from this program will be well versed in quantitative approaches to biology and capable of dissecting complex mechanistic pathways in the living animal. This proposal gains considerable strength by aligning with outstanding UMN gateway graduate programs to markedly broaden and deepen the applicant pool from which we will select outstanding trainee candidates. Moreover, the proposal includes unique tailor-made academic courses in the Physiology of Inflammation and in Computational System Physiology featuring state-of-the-art computational biological and informatics approaches. These courses have been developed expressly for this new training program. Here Ph.D. trainees will gain insight into to the most urgent problems today in human patients with metabolic and cardiovascular medical disorders. Trainees will gain further experiential training in physiological research via conferences, seminars, symposia, retreats, journal clubs, and group meetings with program faculty, including physician-scientists. This program is designed to enable our graduates to pursue unique career pathways, spanning from academia to bio-industry in systems and integrative biology fields of research.

NIH Research Projects · FY 2026 · 2023-04

Abstract Virus-specific CD8 T cells exert potent antiviral activity against HIV-1 and SIV. Despite abundant CD8 T cell responses, these are unable to fully suppress virus replication. This is likely due to the majority of viral replication occurring in CD4+ T cells concentrated in specific areas in the body, such as B-cell follicles in secondary lymphoid tissues and in intestinal tissues. Viral reservoir studies indicate that the majority of SIV and HIV replication may occur in the intestinal mucosa: After ART interruption, the location, abundance, and phenotype of recrudescing virally infected cells are not well understood. However, emerging data suggest that both CD4 T cells, as well as myeloid cells, in lymph node (LN) and the intestine are important contributors. Thus, there is a need to develop methods to reduce viral load in mucosal tissues, as well as in LN, and to more fully understand the source of viral reservoirs in non-lymphoid tissues. CAR T cells can be potent mediators of immune control and these are being explored as an HIV therapy. We have developed and begun testing an HIV/SIV targeting CAR. In this study, we have developed and propose to test autologous T cells engineered to express the HIV-targeting CAR and CCR9, a molecule that will target cells to the intestinal mucosa. We will test our hypothesis that targeting HIV-specific T cells to the intestinal mucosa, as well as B cell follicles, will lead to durable remission of HIV. We propose two aims to test the hypothesis. Aim 1: to determine the in vivo localization, persistence and antiviral efficacy and impact on intestinal mucosa, lymphoid tissue, and other tissue viral reservoirs in SIV infected ART suppressed rhesus macaques infused with autologous CAR/CCR9 T cells. Aim 2: to determine whether combination therapy using both intestine- and lymphoid follicle-homing CAR T cells is superior in promoting sustained remission of SIV after antiretroviral drug treatment interruption. If successful, this work may lead to an immunotherapy that leads to long-term suppression of HIV.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY/ABSTRACT Neuropsychiatric (mental, behavioral and neurological) disorders are increasingly dominating the burden on US healthcare. Yet, our understanding of such disorders is largely restricted to a description of symptoms, and the treatments remain palliative. Several large-scale efforts, including the Human Connectome Project (HCP) and the BRAIN Initiative call for the development of technologies to map brain circuits to improve our understanding of brain function. Magnetic resonance imaging (MRI) plays a central role in these initiatives as a powerful non-invasive methodology to study the human brain, including anatomical, functional and diffusion imaging. Yet, MRI methods have major limitations on achievable resolutions and acquisition speed. These affect both high resolution whole brain acquisitions that aim to image voxel volumes that contain only a few thousand neurons for improved understanding of the brain, and also the more commonly utilized research and clinical protocols. This, in turn, necessitates improved reconstruction methods to facilitate faster acquisitions. Several strategies have been proposed for improved reconstruction of MRI data. Recently, deep learning (DL) has emerged as an alternative for accelerated MRI showing improved quality over conventional approaches. However, it also faces challenges that hinder its utility, especially in high-resolution brain MRI, including need for large databases of reference data for training, concerns about generalization to unseen pathologies not well-represented in training datasets, robustness issues related to recovery of fine structures, and difficulties in training networks for processing multi-dimensional image series. In this proposal, we will develop and validate robust and efficient learning strategies for high-resolution brain DL MRI reconstruction without large databases of reference data. We will develop self-supervised learning methods for training with small referenceless databases or in a scan-specific manner. We will augment these with uncertainty-guided training strategies for improved recovery of areas with high uncertainty, methods for synergistically combining random matrix theory based denoising with DL reconstruction, and memory-efficient distributed learning techniques to process large image series. Our developments will enable at least a two-fold improvement in acceleration rates over existing protocols, and at higher resolutions. They will be validated on HCP-style acquisitions with extensive anatomical, functional and microstructural evaluation at multiple resolutions. Finally, we will curate a whole brain sub-millimeter HCP-style database for studying functional and structural connectivity at the level cortical layers and columns, while also facilitating technical developments for new modeling, image processing and reconstruction algorithms. Successful completion of this project has the potential to transform the scales that can be imaged with MRI, improve the quality of existing protocols and/or significantly reduce scan times, leading to reductions in healthcare costs, improved diagnosis and/or increased patient throughput.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY/ABSTRACT Drug overdose deaths continue to rise in the United States, accounting for over 100,000 deaths in 2021 alone. Methamphetamine and opioid co-use are largely fueling the most recent increase in mortality. In addition, co- use associated hospital admissions increased more than five-fold between 2003 and 2015. Previous research and hospital-based addiction services for substance use have focused on opioid use alone. These addiction medicine services have increased rates of medication for opioid use disorder and improved patient linkage to continuing outpatient treatment. As a result, hospital length of stay and readmission rates have decreased. However, past research indicates individuals with co-use have different sociodemographic and health profiles and are less likely to receive medication for opioid use disorder than individuals who use opioids alone. There is limited evidence of how existing hospital-based addiction services effect patients who co-use methamphetamine and opioids or how these services could be tailored to better treat hospitalized patients with co-use. The overall objective of this Kirschstein-NRSA F30 fellowship is to generate evidence that can guide the eventual development, implementation, and evaluation of hospital addiction medicine treatment services tailored to co-use. The specific aims of this proposal are: 1) to characterize patient demographics and hospitalization characteristics associated with co-use from a nationally representative sample of inpatient hospitalizations, 2) to quantify the risk of re-admission and mortality following hospital discharge, by substance use profile, and whether differences vary by the receipt of hospital-based addiction services and 3) to identify barriers and facilitators to providing hospital-based addiction services tailored for patients with co-use. The proposed mixed methods project is based on a sequential explanatory approach. It includes training in both quantitative (Aims 1 & 2) and qualitative (Aim 3) research methodologies. Through this work and a thoughtfully designed training plan, the trainee will achieve the following goals: 1) develop health services research expertise, 2) advance the substance use research field as it relates to polysubstance use, 3) integrate research, clinical, and advocacy activities in the field of addiction medicine. This supports his long-term goal of bridging health services research and clinical medicine to support patients with substance use disorders as a physician-researcher.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY: Osteonecrosis of the femoral head [ONFH] is a crippling hip disorder affecting children and young adults that can lead to collapse of the femoral head, osteoarthritis, and the need for a hip replacement at a young age. Clinical management of young patients with early-stage ONFH focuses on preventing femoral head collapse, at which point little can be done to salvage the hip joint. However, one of the primary hip preservation treatments for early-stage ONFH, core decompression surgery (which involves drilling into the femoral head), fails to prevent disease progression in 30% or more of patients. This failure can be attributed in part to delayed diagnosis and limited ability to predict and monitor treatment response using current clinical imaging methods (radiography and conventional T1- and T2-weighted magnetic resonance imaging [MRI]). Our long-term goal is to improve clinical outcomes for children and adults with or at risk for ONFH through the synergistic advancement of imaging and therapies. The objective of this R01 proposal is to address the need for better imaging to inform treatment of early-stage ONFH by characterizing the sensitivity of advanced MRI techniques to assess bone ischemia, necrosis, and repair. Our central hypothesis is that quantitative MRI methods are sensitive in detecting early-stage ONFH and provide quantitative measures of the degree of ischemic injury and subsequent repair to the femoral head. We will build upon our promising work using a piglet model of ischemic ONFH, which has demonstrated that quantitative, non-contrast- enhanced MRI techniques, including relaxation time mapping, diffusion imaging, and perfusion imaging, are sensitive and reliable in detecting ischemic injury to the femoral head. In Aim 1, we will expand upon our piglet model studies to identify the cellular changes driving the sensitivity of the quantitative MRI methods to ischemic injury and drilling-induced repair of the femoral head. Animals will be imaged in vivo at 3T MRI, and the femoral heads will subsequently be assessed histologically. In Aims 2 and 3, we will take initial steps to clinically translate the quantitative MRI methods by conducting pilot studies to detect early-stage injury and monitor response to core decompression treatment in patients with ONFH. In Aim 2, we will evaluate whether the quantitative MRI methods can detect femoral head ischemia without use of a gadolinium contrast agent in a pilot study of children with juvenile idiopathic ONFH (also known as Legg-Calvé-Perthes disease). In Aim 3, we will evaluate whether the quantitative MRI methods can detect a reparative response to core decompression in a pilot study of adults being treated for early-stage ONFH, and whether the response differs in those patients whose treatment fails to prevent femoral head collapse versus those whose treatment is successful in preventing disease progression. Collectively, these aims are a critical step toward improving long-term outcomes for young patients with early-stage ONFH through advancement of imaging to allow for new opportunities for therapeutic intervention and the evaluation of the efficacy of new treatments.

NIH Research Projects · FY 2026 · 2023-03

Modified Project Summary/Abstract Section Both genetic and environmental factors contribute to the development of Type 2 diabetes (T2D). Hyperinsulinemia is commonly seen among pregnant women with prediabetes, obesity, and gestational diabetes, and their offspring has a greater risk for developing T2D. Yet, no current study addresses the long-term/longitudinal metabolic outcomes of the offspring when the mother is hyperinsulinemic. Furthermore, the mechanistic link between maternal hyperinsulinemia and the programming of metabolic disease in the offspring remains largely unknown. The dogma is that insulin does not cross the placenta into the fetus to regulate fetal growth. However, maternal insulin can act as a growth factor and an anabolic hormone binding to the placental insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF1R) to drive critical placental function, including nutrient flux to the fetus. Thus, maternal insulin can change placental function by altering nutrient availability to fetal metabolic tissues causing permanent changes that predispose the offspring to T2D in adulthood. Indeed, we have compelling preliminary data showing increased body weight and glucose intolerance in the offspring of hyperinsulinemic dams. We identified that placental-specific IR deletion has a beneficial effect in improving glucose tolerance in the offspring of hyperinsulinemic dams. These observations provide a strong premise that the placenta integrates maternal hyperinsulinemia signals with placental nutrient flux to the growing fetus, thereby programming the metabolic health of the offspring. In this grant, we will test the main hypothesis that the increased body weight and glucose intolerance programming in the offspring by maternal hyperinsulinemia is caused by increased placental nutrient flux to the fetus, which is mediated by increased IR and IGF1R signaling and their downstream targets, mTOR and GLUT4, in the placenta. To test this hypothesis, we developed new innovative models of maternal hyperinsulinemia with or without placental IR or IGF1R deletion, to leverage and obtain a detailed in vivo mechanistic approach of metabolic and physiological studies in a longitudinal cohort of offspring. In Aim 1, we will define long-term metabolic outcomes and signaling mechanisms whereby maternal hyperinsulinemia regulates metabolic health of the offspring using functional studies with preclinical genetic models of maternal hyperinsulinemia during pregnancy with or without placenta-specific loss of IR, IGF1R, IR/IGF1R compound or mTOR. In Aim 2, we will determine maternal-to-fetal nutrient flux in the offspring with or without placenta-specific insulin-signaling or GLUT4 deficiencies. These mechanistic studies are highly significant because they will define the molecular mechanisms whereby maternal hyperinsulinemia impacts metabolic health, and they underscore the importance of clinically controlling insulin levels during pregnancy, similar to glucose, to improve pregnancy outcomes. Thus, the anticipated success of this project will have significant implications in improving women’s reproductive health and the metabolic health of the offspring.

NIH Research Projects · FY 2026 · 2023-02

ABSTRACT Nearly one-third of deceased donor livers are unused for transplant or other purposes. Many of these organs would be valuable for therapeutic and research applications if preservation times could be extended. Cryopreservation at ultralow temperatures (< -140°C) can enable indefinite organ storage. Previous attempts at organ cryopreservation have failed due to cellular and structural disruption caused by ice crystal formation. One promising approach that overcomes the limitations of conventional strategies is vitrification – that is, cooling organs so quickly that the water within the organ cannot undergo the phase transition from liquid to solid ice. With the help of cryoprotective agents (CPAs), the organ enters a stable glass-like state wherein the viable storage time is theoretically unlimited. The critical challenge, however, is rewarming without ice formation or cracking. If rewarming is too slow, ice crystals form; if rewarming is not uniform, thermal stress causes cracking. Hence, speed and uniformity of warming are essential. We have developed a novel rewarming approach termed “nanowarming” that achieves both objectives. Iron oxide nanoparticles are perfused throughout the vasculature of the organ along with CPA solutions. The organ can then be vitrified by cooling (an existing technology) and rewarmed as needed by placing it in a radiofrequency coil that induces heating in the nanoparticles and, therefore, from within the organ. In preliminary studies, for the first time we have shown that we can vitrify and nanowarm human sized (i.e. porcine) and rat livers, thereby avoiding ice formation or cracking and preserving viability and organ-level function. Following on this physical success, we propose here the first study to assess both transplantation and biological viability/function success of these nanowarmed organs. Our central hypothesis is that cryopreservation by vitrification and nanowarming will enable functional long-term whole human liver banking for transplant, therapeutic and biomedical research purposes. While our long-term goal is to develop a method for cryopreserving human livers for transplant, our goals for this project are to refine whole-liver preservation technology to 1) improve in vitro and in vivo functionality of preserved rat livers during normothermic perfusion and in transplant models, 2) determine the mechanisms of cellular stress, injury, and death resulting from liver cryopreservation and strategies for injury mitigation, and 3) provide further investigation of large animal (porcine) and human liver cryopreservation and rewarming. If successful, this approach could revolutionize how these precious resources are allocated and utilized for patient and societal benefit.

NIH Research Projects · FY 2025 · 2023-02

Project Summary This research plan is aimed at creating an off-the-shelf transcatheter vein valve capable of regeneration. If successful, it could treat many thousands of patients who suffer from chronic venous insufficiency in deep veins, untreatable with compression stockings, and the sequelae of debilitating leg ulcers, as there is currently no FDA-approved prosthetic vein valve. This novel vein valve is created from a tissue grown from donor dermal fibroblasts directly on a nitinol stent followed by decellularization. We have shown in a recent publication that the resulting bileaflet valve meets hydrodynamic performance criteria in vitro and regenerates with host cells, including endothelialization, without gross calcification, stenosis, or thickening of the leaflets post-delivery in the ovine iliac vein model based on initial testing. We propose here to make a key improvement upon the initial design and results by using a stent that transitions from a circular section to an oval section, emulating vein valve sinuses to improve hemodynamics and mitigate leaflet fusion to the valve wall that currently occurs in vivo over time. In this milestone-driven plan, valve geometries will first be screened using computational modeling of valve function and then characterized hydrodynamically. Valves of 12 mm diameter enhanced with these sinuses will be delivered via catheter to the ovine iliac vein for up to 24-week duration with (R61 phase) and without (R33 phase) sustained anti-coagulation based on the rate of endothelialization. Delivery will be conducted in the normal anatomy and in a venous reflux model in sheep achieved by compromising the tricuspid valve sufficiently to ensure the valves are cycled in these quadrupeds. Longitudinal assessment of valve function will be made via ultrasound and venogram. Harvested valve leaflets will be assessed for dimensions, tensile mechanical properties, cellularity and phenotype via immunohistochemistry, and matrix composition via biochemical assay and histology. An INTERACT meeting will also be conducted during the R33 phase to ascertain GMP manufacturing and GLP testing expectations of the FDA for a transcatheter vein valve, extending prior FDA interaction regarding the decellularized tissue that we create in vitro to include stent and valve testing in light of ISO standards that exist for transcatheter heart valves. Our Office of Technology Commercialization will also be engaged regarding patent protection beyond the PCT filed and commercialization opportunities.

NIH Research Projects · FY 2026 · 2023-02

Summary The biochemical hallmark of FKRP-associated dystroglycanopathies is the hypoglycosylation of α-dystroglycan (α-DG), which leads to disruption in the interaction of α-DG with extracellular matrix proteins, ultimately leading to muscle wasting. Recessive mutations in FKRP are associated with a heterogeneous spectrum of muscle disorders, ranging from severe early-onset to mild late-onset limb-girdle muscular dystrophy (LGMD2I) to several forms of congenital muscular dystrophy (MDC1C), including severe Walker-Warburg Syndrome. Respiratory impairment due to loss of diaphragm function is a prominent complication of both LGMD2I and MDC1C. No approved therapy currently exists for dystroglycanopathies. There has been tremendous excitement for the therapeutic potential of reprogrammed induced pluripotent stem (iPS) cells in treating genetic diseases. The premise of this project is that stem cell-based therapy consisting of human skeletal myogenic progenitors derived from iPSCs will replenish diseased muscle with normal functional muscle fibers as well as muscle stem cells, which have the potential to provide long-term therapeutic effect in dystroglycanopathies. We have developed and extensively validated a method to generate engraftable skeletal myogenic progenitors from pluripotent stem cells through conditional expression of Pax3 or Pax7. This approach results in highly efficient generation of therapeutic myogenic progenitors, which when transplanted into dystrophic mice locally or systemically produce large quantities of functional skeletal muscle tissue that incorporates normally into the host muscle. Importantly, a fraction of transplanted cells remains mononuclear, and displays key features of skeletal muscle stem cells, including satellite cell localization, response to re-injury, and contribution to muscle regeneration in secondary transplantation assays. Therefore, our technology comprises a cell therapy to rebuild functional skeletal muscle, robust to future damage, in hosts with muscular dystrophy. We have recently shown that mouse and human PSC-derived myogenic progenitors contribute to significant myofiber and satellite cell repopulation in the immunodeficient FKRPP448L-NSG mouse model that we generated. Of therapeutic relevance, we have evidence of successful delivery of these myogenic progenitors directly into the diaphragm of FKRP mice. In addition, we have developed a universal gene correction strategy for FKRP, applied this to patient-specific WWS and LGMD2I iPSCs, and demonstrated in vitro and in vivo rescue of functional α-DG glycosylation. In this application, we propose studies that are critical for the development of successful therapeutic approaches for dystroglycanopathies, including understanding 1) the effect of the environment on the engraftment of transplanted cells and 2) the long-term functionality and molecular characteristics of human gene edited WWS and unaffected iPSC-derived myogenic progenitors, important for both autologous and allogeneic future therapeutic applications, respectively.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY/ABSTRACT This proposal for the NIH Pathway to Independence Award (K99/R00) focuses on the training of Dr. PingHsun Hsieh to become an independent investigator of large-scale genomics and human population genetics. Dr. Hsieh is a population geneticist by training, and the proposed studies will advance his training into long-read- based sequencing technologies and novel machine-learning approaches to study the fitness consequences of new mutations, with a focus on structural variants (SVs), in humans and nonhuman primates. Another essential piece will be the development of resources on which types of new SVs are most likely to be pathogenic and hence most worth further effort by medical researchers. The methods developed in this work will enable other researchers to do more hypothesis-free analysis of SVs in disease etiology. Specifically, the training program will center on the study of the distribution of fitness effects of new SVs in human and nonhuman primates using high-quality SV calls and genotypes from several large-scale long- and short-read sequencing projects. The mentored work will take place under the supervision of the primary mentor, Dr. Evan Eichler, and the co-mentor, Dr. Sharon Browning, both at the University of Washington (UW). The mentor and co-mentor are well-established experts in the characterization of genomic variations using high-throughput technologies and the development of stochastic modeling methods for large-scale genetic data, respectively. Dr. Hsieh will also gain advice from a formal advisory committee as well as through activities arranged by the Department of Genome Sciences (GS), which is an optimal place for the mentored training providing the candidate with access to outstanding scientists in areas including genetics of model organisms, disease, population genetics, and the development of high-throughput genomic technologies. While found in nature and yet generally deemed to be deleterious given their size, SVs can be beneficial, and thus, the distribution of fitness effects (DFE) of new SVs (i.e., the relative frequencies of beneficial, neutral, and deleterious SVs) remains elusive. In the proposed studies, we will infer the DFE of new SVs and other variants to assess their relative importance in nature, which in turn helps prioritize variants (e.g., SVs vs. single- nucleotide variants [SNVs]) in medical genetics. Specifically, in the K99/R00 phases we will (1) infer the DFE of new SVs and SNVs using a diverse panel of ~100 long-read and ~4,000 short-read high-coverage human and nonhuman primate genomes; (2) compare the DFE of new mutations among primates using contemporary and ancient DNA genomes; and (3) study the fitness effects and selective constraints on diseases in different mutation categories in large cohorts of >20,000 genomes. The skills learned in this proposal are on the cutting-edge and are tailored for the candidate to amass a great amount of knowledge in new areas of genomics, which will be applicable to many organisms and diseases and critical to the candidate’s future independent laboratory.

NIH Research Projects · FY 2026 · 2023-02

Project Summary Cancer care is becoming increasingly complex, with growing demands on patients’ energy and time. As a result, patients often have to neglect their usual life activities and relationships. Such ‘time toxicities’ of cancer are rarely acknowledged and play little role in care considerations. Further, no scoring systems exist to measure the time burden of cancer care. Having to weigh potential survival benefits of treatments against added time burdens especially affects those at risk of premature death. Therefore, there is a critical need to measure and reduce the time toxicities of cancer. The overall objectives of the proposed research are to describe and quantify sources of cancer-related time toxicity among individuals receiving treatment for cancer and their effect on well-being, and to create time toxicity scores which can be used in future studies to identify opportunities to minimize time toxicity. The general hypothesis is that combining sensor-based objective data with subjective self-reported measures of time spent on healthcare-related activities will accurately measure the time burden of cancer care and identify areas for interventions related to treatment delivery to reduce this burden. This hypothesis will be tested via the following specific aims: (1) Measure and describe components of objective time use associated with cancer-related healthcare interactions via a mobile health application; (2) Characterize associations between measures of cancer-related time use and self-reported well-being, and explore the role of context in modifying these associations; (3) Create a time toxicity summary score based on measures of cancer-related time use and assess its association with psychosocial outcomes. We will conduct a 28-day prospective cohort study of 80 individuals with advanced stage ovarian cancer or metastatic breast cancer. Using an existing smartphone application, we will automatically track time spent on daily cancer activities, augmented by participant-reported details on specific activities, well-being (daily), and quality of life (baseline and end of study). We will estimate associations between objective time use and daily well-being, explore variations in these associations by patient characteristics, develop a multidimensional scoring system of time toxicity differentiating between episodic toxicity, travel toxicity, opportunity toxicity, and scheduling toxicity, and measure the associations of these scores with patient reported psychosocial outcomes. Upon completion we will have developed objective measures of daily time use by cancer activity and patient characteristics (Aim 1), estimated associations with daily well-being and explored variations by patient characteristics (Aim 2), and developed a multidimensional time toxicity scoring system (Aim 3). The proposed research is innovative because time toxicity is a novel concept within cancer survivorship which we will capture combining objective sensor data with self-report data. This study will have a significant impact because time matters to patients and our measures of cancer care time burdens will facilitate future interventions to reduce time toxicity.

NIH Research Projects · FY 2026 · 2023-02

Lipid droplets (LDs), the organelles responsible for lipid storage and the largest energy reserve in most cell types, are the defining characteristic and etiological factor in the development of non-alcoholic fatty liver disease (NAFLD). Moreover, LDs are recognized to play central roles in coupling NAFLD to more systemic comorbidities such as Type 2 Diabetes and cardiovascular disease among others. LDs interact with numerous organelles, especially ER and mitochondria, which are thought to coordinate de novo LD biogenesis and fatty acid (FA) transfer/oxidation, respectively. However, published work from our laboratory and others have questioned the established dogma that direct transfer of FAs from LDs to mitochondria is the primary route of their oxidation during fasting. Using a multifaceted approach involving organelle proteomics, isotope tracing, and numerous super resolution microscopy approaches, we show for the first time that in the liver, the proteomes and metabolism of mitochondria attached to LDs (peridroplet mitochondria, PDM) support lipid anabolic pathways, whereas mitochondria unattached to LDs (cytosolic mitochondria, CM) a geared for enhanced FA oxidation and OXPHOS. Moreover, our data point to an important role for mitochondrial-associated membranes (MAMs), an ER domain that tightly interacts with mitochondria, as a key component in regulating LD-mitochondria interactions and dynamics. Collectively, these data suggest that interactions with LDs profoundly affect organelle dynamics and function. Based upon these data, the objective of this application is to define how LD interactions affect lipid metabolism and sensing to coordinate ER and mitochondrial function under physiological and pathophysiological conditions. We hypothesize that interactions of LDs with ER and mitochondria are critical modulators of MAM lipid sensing and mitochondrial function that govern hepatic lipid and energy metabolism. To test this hypothesis, we propose the following three specific aims: Aim 1 - To comprehensively define LD- mitochondria interactions and their impact on FA trafficking; Aim 2 - To determine the mechanisms through which subpopulations of MAM differentially impact mitochondrial bioenergetics and lipid metabolism; and Aim 3 - To determine how NAFLD impacts LD/MAM/mitochondria dynamics in NAFLD. To complete these aims, we will employ a wide range of advanced super resolution imaging approaches, proteomics and RNA sequencing, isotope tracing, and other cell biology and biochemical approaches in cells, mouse models and human liver biopsies. Upon completion of these studies, we will expect that we will have revealed novel mechanisms through which LDs can alter cellular function/dysfunction that underlie NAFLD etiology. We anticipate that this work will open new areas of research into intracellular signaling dynamics, which will advance therapeutic approaches targeting NAFLD and related comorbidities.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY/ABSTRACT Osteosarcoma, the most common primary tumor of bone, primarily affects children, adolescents, and young adults. A diagnosis of osteosarcoma is devastating, as approximately half of pediatric osteosarcoma patients experience metastasis and ultimately succumb to the disease within 10 years of their diagnosis. Currently there is no diagnostic test to predict prognosis, so all patients are treated with aggressive surgery and intense chemotherapy with high rates of toxicity. However, a subset of patients may not require as aggressive therapy to achieve remission. Additionally, those that survive have a high incidence of lifelong morbidities, including treatment-related secondary malignancies. Accurate prognostic indicators could be integrated into the standard of care for osteosarcoma to guide therapy. Children with more favorable prognoses could be treated more conservatively, reducing the need for aggressive surgery, and decreasing the intensity of systemic therapy. This would decrease the likelihood and severity of long-term morbidities, and reduce the probability of secondary, treatment-related malignancies without negatively affecting prognosis. Conversely, patients with a worse prognosis could receive more aggressive treatments or be guided to experimental clinical trials to improve their long-term survival. In this project, we will develop a serum exosomal gene signature associated with prognosis in pediatric osteosarcoma. Exosomes are membrane-bound microvesicles containing cargo associated with tumor biology and disease state. We will first identify biomarkers by sequencing serum exosomes from a large cohort of pediatric osteosarcoma patients with known clinical outcomes. We will then identify genes associated with metastatic propensity using xenograft mouse models established from pediatric osteosarcomas with distinct biologic behavior. We will analyze co-regulated gene clusters and apply machine learning, improving sensitivity and specificity, and ultimately resulting in a more robust gene signature. The osteosarcoma gene signature developed in this project can be utilized in the clinical setting to predict prognosis, stratifying patients into more appropriate treatment categories, and having the potential to improve management of this devastating disease. Additionally, these biomarkers will contribute to our understanding of the biological behavior and progression of osteosarcoma, allowing us to infer mechanisms of host response, metastasis, and response to therapy. Importantly, this K01 is critical to advancing my career as a translational scientist by providing the necessary protected time and dedicated resources to perform high-quality, clinically relevant research, under the guidance of an exceptional multidisciplinary mentor team. This award will facilitate my transition to independence, as the data procured in this project will allow me to be competitive for future independent funding applications. Additionally, as I complete these aims, I will develop the necessary knowledge and leadership skills of a successful independent research scientist specializing in exosome biology and translational models of pediatric osteosarcoma and ultimately expanding to other cancers.

NIH Research Projects · FY 2026 · 2023-02

Project Summary/Abstract Primary cilia are multisensory organelles that function as cellular antennae. We found that ciliary defects in cholangiocytes and/or the loss of primary cilia are associated with biliary tract diseases like polycystic liver disease (PLD) and cholangiocarcinoma (CCA). A better understanding of the signaling regulated by cilia and mechanisms of ciliary loss in diseased cholangiocytes is critical to design new therapies based on the restoration of cilia, i.e. ciliotherapies. Our current overall objective is to understand the role of cilia in the regulation of epidermal growth factor receptor (EGFR) signaling. EGFR signaling is abnormally persistent and enhanced in PLD and CCA, two diseases with ciliary dysfunction. Furthermore, we aim to explore the mechanisms of ciliary loss in cholangiocytes – especially how the autophagy machinery is targeted to resorption of this organelle. This proposal will assess how cilia-autophagy communication works in cholangiocytes to reduce ciliary expression and, consequently, how the loss or dysfunction of cilia enhances EGFR signaling. We propose that pathologically-induced ciliophagy accounts for ciliary loss/dysfunction, inducing sustained EGFR signaling. We propose three Specific Aims: In Specific Aim 1: To characterize molecular mechanisms of the ciliary-dependent degradation of activated EGFR, we will assess the need of cilia for activated EGFR degradation; characterize the mechanisms of EGFR translocation to primary cilia; and assess the hypothesis that the E3 ubiquitin ligase c-CBL translocates to the primary cilia upon EGF signaling and drives the degradation of activated EGFR located in the cilia. In Specific Aim 2: To identify the key players involved in targeting ciliary components to the autophagy machinery, we will assess the role of autophagy and HDAC6/SIRT1 in ciliary expression in vitro; assess the role of HDAC6/SIRT1 in ciliophagy in vivo; study the interaction between ciliary proteins and autophagy cargo receptors; and test the hypothesis that in ciliary-defective cholangiocytes, overexpression of deacetylases induces lysine deacetylation of ciliary components, which leads to ubiquitination of the same residues and targeting of the autophagy machinery by specific autophagy cargo receptors. In Specific Aim 3: To test the combination of specific deacetylases, autophagy, and EGFR inhibitors in pre-clinical rodent models as a therapeutic approach, we will assess the effect of HDAC6 inhibition (Tubastatin-A or ACY-1215), and/or SIRT1 inhibition (Sirtinol) in combination with autophagy inhibitors (e.g., HCQ, SAR405) with or without EGFR inhibition (Erlotinib, Afatinib) in vitro and in vivo; assess the in vivo effects of Ciliomax (a novel dual inhibitor we recently developed) plus EGFR inhibition; and assess the most promising treatments in patient-derived xenografts. Impact: identifying novel targets could lead to much-needed new therapeutic strategies for these devastating diseases. Our experiments in in vitro and pre-clinical rodent models will characterize the ciliary-dependent regulation of EGFR and the communication between primary cilia and the autophagy process, which will lay the foundation for potential clinical trials.

NIH Research Projects · FY 2026 · 2023-01

Cerebral aneurysms (CAs) are out-pouching dilations of cerebral arteries caused by local wall weakening and maladaptive remodeling. Though rupture is relatively rare, the post-rupture survival rate is low, due to complications such as vasospasm and stroke. Since the majority of cerebral aneurysms are stable, the ability to predict rupture would both allow early intervention and eliminate unnecessary surgical procedures for stable aneurysms. Many computational models have been developed with the aim of predicting rupture based on correlation with clinically measurable factors, such as aneurysm shape or blood flow dynamics. But, these models are not yet accurate enough for them to have been used in the clinic. A major shortcoming of the current approach is that it does not consider the complex mechanics of rupture but instead tries to leap from shape and/or fluid dynamics directly to rupture risk. In contrast, we will build on our understanding of mechanical heterogeneity and its role in tissue growth, remodeling, and failure. By incorporating heterogeneity into the description of the CA, we will inform future models and enable more accurate assessment of CA rupture risk. We hypothesize that cerebral aneurysms are mechanically heterogeneous, and this heterogeneity is predictive of the rupture potential of the aneurysm. We further hypothesize that the material heterogeneity can be determined from (i) the wall shear stress field caused by blood flow in the aneurysm and (ii) the geometry of aneurysm, both of which can be determined in a clinical setting. We propose a series of novel experiments and computational models aimed at elucidating the role of tissue heterogeneity on cerebral aneurysm growth, remodeling, and rupture. Using freshly excised human aneurysm tissue, we will measure regional tissue-scale mechanical properties, ECM structure and composition, cell organization, and the rupture stress of the aneurysm. Next, we will develop and use computational models to elucidate the biophysical mechanisms that connect tissue properties to aneurysm rupture. Finally, we will use computational analyses of the architecture and blood flow mechanics within the aneurysm to connect these clinically-measurable metrics to clinically non-measurable material properties. The findings from this study will provide key mechanistic insights needed to advance cerebral aneurysm rupture prediction models.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY Defining how FoxP3+ regulatory T cells (Tregs) limit amnestic responses to common allergens has the potential to provide new and effective therapies for asthma and other atopic diseases. Recent work has demonstrated that T helper type 2 (Th2) tissue-resident memory T cells (Trm) persist around the airways following allergic sensitization where they become rapidly activated and drive the asthma phenotype upon allergen re-exposure. The immunoregulatory mechanisms that limit the proinflammatory functions of Th2-Trm at mucosal sites are less clear. The objective of this proposal is to define the factors influencing allergen- specific resident (r) Treg maintenance in the lungs and their suppression of Th2-Trm following re-exposure to inhaled allergen. Our central hypothesis is that allergen-specific rTregs are positioned in a unique niche in the lung that enables potent suppression of allergic inflammation. Mechanistically, we hypothesize that rTregs co- localize with Th2-Trm based upon the CXCR6 and CCR8 chemokine systems. This proposal will explore these questions using novel experimental systems to define the function of rTregs in a mouse model of allergic asthma, including polyclonal adoptive transfer of house dust mite (HDM)-specific cells, parabiosis, single cell RNA sequencing analysis of HDM-specific T cells, and CRISPR-Cas9 gene editing. With Aim 1, we will build upon preliminary data to define the role of allergen-specific rTregs in suppressing lung Th2-Trm. We will characterize the localization, persistence, suppressive potency, and transcriptional profile of the Tregs that persist in the lung following allergen clearance. In Aim 2, we will determine the role of the CXCR6 and CCR8 chemokine systems in rTreg positioning and function. To identify additional factors important for the rTreg tissue residency program and provide additional training in genome editing and systems biology, we will perform an in vivo CRISPR-Cas9 screen on HDM-specific TCR transgenic cells isolated from the HDM memory lung. Dr. Nelson will perform the work in this K08 proposal, sponsored by Boston Children’s Hospital, at the Center for Immunology and Inflammatory diseases (CIID) at Massachusetts General Hospital (MGH) under the mentorship of Dr. Andrew Luster. Dr. Nelson has developed a career development plan consisting of coursework in advanced microscopy, bioinformatics, and translational research, along with mentored hands-on training in cutting-edge research methods to explore his research aims. The goal of this K08 award is to provide Dr. Nelson with the necessary skills and knowledge to become an independent, NIH-funded investigator with expertise in human disease models of atopic diseases and immunoregulation in non-lymphoid tissues.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Drosophila melanogaster is one of the leading animal models for biomedical research, with researchers generating new lines of Drosophila every year for studies in numerous cancers, genetic disorders, and other maladies. Currently there are greater than 160,000 Drosophila stocks held at different stock centers around the world. As reliable and cost-effective approaches for long-term preservation of Drosophila stocks are lacking, individual labs and stock centers must maintain their lines as living populations, which is resource-intensive and puts the stocks at risk of loss. In 2021, our group developed an easily implemented and robust cryopreservation protocol for Drosophila melanogaster embryos which can be applied in any lab without the need for specialized instruments. This protocol is broadly applicable, and it has been successfully used to preserve 25 distinct strains from different sources. For most strains, >50% of the embryos hatch and >25% of the resulting larvae develop into fully functioning adults after cryopreservation and rewarming (normalized survival to control embryos), providing sufficient numbers of adults to revive the strain. To accelerate the cryopreservation and archival of critical Drosophila stocks, we will develop, standardize, and disseminate resources to individual labs and stock centers. Significant efforts will be devoted to developing reliable methods for cryogenic storage and shipping of Drosophila embryos as well as to study if any significant mutagenic changes occur due to cryopreservation. We will be working with multiple groups within the cryobiology and fly community to gather feedback on and refine our approach. With the collaboration of the Bloomington Drosophila Stock Center (BDSC), we will identify and prioritize the stocks that need cryopreservation to prevent their total loss in the event of a disaster or genetic drift. We will also demonstrate the efficacy of our cryopreservation protocol on a large scale with a selected group of stocks identified by BDSC. Finally, we will also estimate the costs for cryopreservation in comparison to the traditional method of Drosophila maintenance to inform future efforts to protect these precious genetic resources. This is a multi-PI project led by investigators with significant experience in Drosophila genetics and cryobiology.

NIH Research Projects · FY 2025 · 2023-01

Project Summary Breast cancer is the most commonly diagnosed cancer in women, with estrogen receptor positive (ER+) breast cancers accounting for 75% of cases. Endocrine therapies directed at blocking ER action are highly effective; however, 40% of women with ER+ tumors develop resistance and progress to metastatic disease. ER+ tumors relapse late, and tumor cells can remain quiescent for years to decades. Progress in the treatment of metastatic breast cancer is limited by strategies that primarily target rapidly proliferating tumor cells. Contributing factors to advanced disease progression include breast cancer stem cells (CSC), which are poorly proliferative and exist as minority populations in therapy resistant tumors. We identified SRC-3 (steroid receptor [SR] co-activator 3) as a novel cytoplasmic binding partner of PELP1. Similar to SRC-3, PELP1 is an ER co-activator, and dynamically shuttles between the nucleus and cytoplasm to act as a nuclear co-activator and cytoplasmic scaffolding protein for growth factor and steroid receptors. PELP1 is primarily nuclear in normal breast, but increased cytoplasmic localization of PELP1 is an oncogenic event that promotes disease progression by unknown mechanisms. We showed PELP1/SRC-3 cytoplasmic complexes drive breast CSC phenotypes and genes associated with pro-survival in ER+ breast cancer models. SRC-3 inhibition disrupts complex formation and cytoplasmic PELP1-induced tumorspheres. Top candidates identified from RNA-seq analysis include PFKFB family members, which are bi-functional kinase/phosphatases that have roles in cancer metabolism and CSC biology. PFKFB3/-4 co-purified with PELP1/SRC-3 complexes; inhibition of PFKFB3/-4 blocked PELP1/SRC-3 complex formation and biology. Remarkably, PELP1/SRC-3 CSC biology is phenocopied in tamoxifen-resistant (TamR) and paclitaxel-resistant (TaxR) models. Herein, we hypothesize that PELP1/SRC-3 complexes amplify signaling inputs to PFKFB family members that mediate altered metabolic pathways required for resistant ER+ tumor cell populations. We will: 1) identify signaling pathways essential for PELP1/SRC-3 driven therapy resistance using mass cytometry, and 2) determine the therapeutic benefits of targeting PELP1/SRC-3/PFKFB complexes in vivo to block cancer progression and metastasis. Our long-term objectives are to identify non-ER therapeutic targets that can be developed as combination strategies to eliminate therapy resistant tumor cells in ER+ breast cancer. During the K22 award, we expect to define the molecular links between cancer cell metabolism and oncogenic events in breast cancer progression, metastasis, and examine the benefits of targeting this pathway to impair late recurrence. This proposal will provide a solid foundation for the candidate’s goal of moving towards translational cancer research during her transition to independence. Delineating the key players will fundamentally redefine standard care options to target therapy-resistant populations in ER+ breast cancer.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Our goal is to establish a novel MRI approach to acquire functional MRI (fMRI) data simultaneously from brain and spinal cord. Unlike standard MRI readouts, our approach does not require a dedicated shimming procedure, as it is based on the newly developed zero echo time MRI pulse sequence entitled Multi-Band SWeep Imaging with Fourier Transformation (MB-SWIFT) which is inherently resilient to magnetic field inhomogeneities. Comprehensive evaluations of the central nervous system (CNS) function will tremendously benefit from this imaging modality, particularly in areas of research such as spinal cord injury, neurodegenerative diseases, pain and aging. Thus far, functional neuroimaging of the CNS has been an untapped area of research due to several technical challenges, the biggest of which is the need to efficiently shim a field of view large enough to cover both brain and spinal cord. Solutions of per-slice dynamic shimming approaches have been proposed. However, they are limited by the settling-time of eddy currents, they considerably prolong the experimental session, and they have been applied only to cover cervical (but not lower) spinal cord. Furthermore, dynamic shimming allows only inefficient sequential rather than simultaneous acquisitions of brain and spinal cord, posing additional challenges for CNS fMRI. As proven by our preliminary data, MB-SWIFT can instead image two fields of view (FOVs) at distant locations in brain and lumbar spinal cord in a true simultaneous fashion (i.e., within 1 ms of each other), thus allowing unprecedented functional imaging of the CNS that is unattainable with standard imaging modalities for fMRI. The current project is a proof-of-concept study conducted in rats, designed to first optimize the MB-SWIFT protocol including the dual RF coil and the dual FOV acquisition, then to demonstrate that fMRI of CNS with MB-SWIFT provides robust fMRI outcomes in absence of dedicated shimming solutions. Moreover, we will optimize spoke order of MB-SWIFT and post-processing pipeline for handling physiological noise, and demonstrate that MB-SWIFT provides surrogate markers of neuronal activity in spinal cord. Finally we will establish inter-subject, intra-subject and inter-site reproducibility of task-based and resting-state fMRI outcomes extracted from the CNS with dual FOV MB-SWIFT. Once completed, the study will provide an invaluable tool for pre-clinical research, and will set the stage for translation to humans.

NIH Research Projects · FY 2026 · 2023-01

Project Summary The mesocorticostriatal network is central to adaptive reward processes, as well as diseases of dysfunctional learning, motivation, and cognition. As learning progresses, there is a transition from initial reliance on nucleus accumbens (NAC) to later recruitment of dorsolateral striatum (DLS), which drives a shift from goal-directed behavior to more automatic behaviors characterized by rapid, stimulus-driven movement sequences. This transition is thought to involve a network of recurrent loops, including dense dopaminergic (DA) input from the ventral tegmental area (VTA)/substantia nigra (SNC) and glutamatergic input from cortex. Critically, however, the contributions of these loops have not been directly tested, and the circuit mechanisms driving this fundamental neurobiological adaptation remain undefined. In the proposed studies, we will use several innovative approaches to investigate how different loop systems within the mesocorticostriatal network communicate in vivo to organize conditioned behavior, and transition from ventral to dorsal striatal control, across learning. One influential anatomical framework, the mesostriatal “spiral” hypothesis, suggests that during learning, information flows serially across a subcortical loop, from the VTA to nucleus accumbens to SNC, to dorsolateral striatum. Despite broad acceptance in the field, support for the striatal spiral has not been demonstrated in vivo. Instead, emerging evidence suggests an alternative hypothesis: that cue-reward learning engages progressive recruitment of the dorsal striatum via nigro-thalamo-cortical circuits, rather than the classic striatal spiral mechanism. In Aim 1 we will combine fiber photometry recordings of somatic DA neuron activity in TH-cre rats with simultaneous recordings of a DA biosensor in the striatum, to characterize the spatial and temporal pattern of information flow through four nodes in the striatal DA system during cue-reward (i.e., Pavlovian) learning, testing predictions from the spiral framework. In Aim 2, we will use trans-synaptic targeting, optogenetics, and photometry to test the ascending spiral framework in vivo, determining if NAC direct pathway neurons disinhibit SNC DA neurons. We will use D1-cre rats to investigate the function of direct pathway output neurons in the NAC and DLS at different stages of learning. Finally, In Aim 3 we will combine photometry recordings of corticostriatal and thalamostriatal circuits with optogenetic manipulation of DA neurons in TH-cre rats, to assess the ability of nucleus accumbens DA signaling to engage the nigro-thalamo- cortical loop. We will then optogenetically manipulate input-defined nigral neurons projecting to the thalamus to determine the functional role of the nigro-thalamo-cortical loop in learning. These studies will resolve longstanding questions about the circuit mechanisms of information flow across striatal input-output circuits, establishing a normative framework for the in vivo functional architecture of the mesocorticostriatal network.

NIH Research Projects · FY 2026 · 2022-12

Project Summary/Abstract Understanding and mitigating the health impacts of climate change and other environmental exposures requires information on population and agricultural characteristics at the community level—a key scale for decision- making and action. Population and housing censuses and agricultural censuses provide in-depth, community- scale data on demographics, education, employment, and living conditions as well as on farming inputs, practices, and productivity. Currently, these data are difficult to access and use for research and decision- making. While nearly every country conducts regular censuses, the results are published independently by hundreds of statistical agencies, often in reports designed for reading rather than for analysis. The proposed project—the IPUMS International Historical Geographic Information System (IPUMS IHGIS)—will assemble these data in an analysis-ready, standardized, and fully documented collection and make them freely available via a user-friendly web-based interface. These data will allow researchers to better answer a wide range of environmental health questions, such as what population-level factors contribute to the spread of vector-borne disease or where people are particularly vulnerable to food insecurity. The project has four Aims: (1) Ingest population and agricultural census data. Using software developed with NSF funding, data and boundaries for approximately 25,000 additional tables from 250 censuses will be added to the IHGIS database, a ten-fold increase to the current collection. Data from countries most vulnerable to health impacts from climate change and providing the finest geographic detail available will be prioritized. (2) Enable linkages between census data and health data, allowing users to attach community-level census data to individual-level health survey data from the Demographic and Health Surveys, Performance Monitoring for Action surveys, and Multiple Indicator Cluster Surveys. The project will also develop a utility to provide geographic unit codes for user-uploaded coordinates, so researchers can link their own health data to IHGIS contextual data. (3) Improve standardization and integration across datasets to support comparisons across time and between countries. Enhancements to the software will enable filtering and selection based on topics and level of geographical detail, standardize units of measurement, and create integrated versions of selected data tables. (4) Support and expand the IHGIS user base through online tutorials, virtual and in-person workshops, webinars, and one-to-one user support to develop a broad community of new and established environmental-health researchers using IHGIS data. Outreach efforts will focus on reaching, training, and supporting a diverse—in background, discipline, and career stage— community of environmental health researchers. By providing rich, detailed, and expansive information on characteristics of populations around the world, IHGIS data will also further the NIEHS goals of developing a program in global environmental health and understanding health disparities and risks for vulnerable populations.