Washington University

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY In this K01 proposal, Dr. Arin Oestreich, PhD, presents a detailed career development plan that will culminate in an independent academic faculty position. Dr. Oestreich will obtain critical skills in (1) animal models of osteoarthritis (OA), (2) chromatin profiling and transcriptomic analysis using single nucleus sequencing, and (3) bioinformatic approaches to integrate multiomic platforms and decipher signaling pathways programed by maternal obesity. Dr. Oestreich uniquely combines her background in maternal obesity with new training in OA to enhance her scientific career. Equipped with this unique dual expertise and preliminary data, Dr. Oestreich will be well positioned to develop an independent research program focused on studying the effects of specific components of the maternal obese milieu on offspring joint health. The scientific goal of this project is to determine the impact of maternal n-6 HFD on the epigenetic regulatory mechanisms governing offspring OA risk and to isolate the effect of maternal dietary n-6 fatty acids on programming offspring OA. Aim 1 will test the hypothesis that maternal n-6 enriched HFD increases the severity of injury-induced OA in the adult offspring and determine the critical window of developmental exposure. Aim 2 will test the hypothesis that maternal n-6 HFD directly programs the knee joint by stably altering the epigenetic landscape of the musculoskeletal progenitors during development. This aim will use multiomic single nucleus sequencing of the epigenome and transcriptome to pursue genetic targets are epigenetically regulated by maternal obesity and known to heighten OA severity in the adult knee. Aim 3 will test the hypothesis that controlling the n-6:n-3 fatty acid ratio in obese dams will decrease maternal-fetal inflammation and protect the adult offspring from developing OA. By determining the impact of specific maternal dietary fatty acids on fetal limb development, the data collected in this proposal will be invaluable for formulating prenatal vitamins with optimal ratios of n-3 PUFAs as a novel strategy to protect the adult offspring joint health. With the support of this K01 and the training provided in her career development plan, Dr. Oestreich will be uniquely poised to attain her primary goal of becoming an independent researcher in the field of developmental programming of musculoskeletal disease.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY / ABSTRACT This is a new application seeking support for the training program in hematology-oncology in the Department of Pediatrics at Washington University School of Medicine. This program had been continuously funded between 1999 and 2020. We have extensively redesigned the program to enhance its flexibility in order to keep pace with the changing landscape of biomedical science. The long-term goal is to produce independent investigators capable of making important contributions to our field. The program is open to MD’s or MD/PhD’s and will support one second-year (PGY-4) and one third-year fellow (PGY-5) each year. The trainees will be recruited from the extensive network of pediatric subspecialty training programs at The at Washington University School of Medicine. The fellowship program is comprised of three to four years of training, only two of which will be supported by the T32. The Hematology-Oncology fellowship includes many qualified applicants with backgrounds in hematology and oncology research. Only those applicants with the highest likelihood of success as physicians/scientists will be chosen for funding. Support decisions will be made by the program leaders. Supported trainees will participate in didactic seminars, lectures and journal clubs that cover genomics, epigenomics, developmental biology, cellular therapeutics, clinical trial design and performance, and survivorship as appropriate to their research projects. Trainees may perform their postdoctoral research in a wide range of laboratory and clinical settings at Washington University, provided the research is relevant to the field of pediatric hematology-oncology. Training will be overseen by a program steering committee with expertise in each of these domains. Drs. Joshua B. Rubin, MD, PhD and Jorge Di Paola MD, PhD will serve as Program Directors and Dr. Laura Schuettpelz, MD, PhD will serve as the Training Director. Support for this of program will provide trainees with the opportunity to leverage the prodigious strengths of the Washington University research and clinical enterprises, in their quests to become the next generation of leaders in pediatric hematology-oncology research.

NIH Research Projects · FY 2026 · 2022-04

Project Summary Therapeutic expansion or activation of brown adipose tissue (BAT) has a potential to be an effective treatment for obesity. BAT, as well as the closely-related beige fat, are characterized by their abundance of mitochondria, which are involved in thermogenesis through uncoupled respiration. In addition to the importance of mitochondrial functions, mitochondrial morphology plays a critical role in thermogenesis. Mitochondria are highly dynamic organelles that continuously undergo cycles of fission and fusion. Adrenergic stimulation-induced mitochondrial fission in BAT promotes uncoupled respiration and thermogenesis. Like mitochondria, peroxisomes are enriched in BAT. Our recently published studies indicate that peroxisomes play a critical role in thermogenesis through their ability to regulate cold-induced mitochondrial fission. The defect in mitochondrial fission and thermogenesis in mice with adipose-specific knockout of the critical peroxisomal biogenesis factor Pex16 (Pex16-AKO) could be rescued by dietary supplementation of peroxisome-derived lipids called plasmalogens. This project seeks to understand the molecular mechanism of peroxisomal regulation of mitochondrial dynamics and thermogenesis. Our preliminary data suggest that norepinephrine stimulation, which activates mitochondrial fission, promotes recruitment of peroxisomes to mitochondria. To understand the role of peroxisomes in mitochondrial dynamics, we performed protein mass spectrometry on mitochondria isolated from BAT of Pex16-AKO and control mice. TMEM135, a peroxisomal and mitochondrial membrane protein, was identified as the most dramatically decreased protein in the knockout BAT mitochondria, with no change of its levels in whole tissue lysates, suggesting that the protein is mistargeted in the absence of peroxisomes. TMEM135 expression in BAT increases with cold exposure. Its knockdown in brown adipocytes results in tubular mitochondria, while the overexpression promotes mitochondrial fragmentation. We hypothesize that peroxisome- mitochondria membrane contacts regulate mitochondrial localization of TMEM135 in a plasmalogen-dependent manner and that TMEM135 mediates mitochondrial fission to promote thermogenesis. We propose three specific aims to test this hypothesis. The first aim will define the role of TMEM135 in mitochondrial dynamics and function in brown adipocytes. The second aim will determine if TMEM135 overexpression in mice promotes energy expenditure through increased BAT mitochondrial fission and if it rescues thermogenesis in Pex16-AKO mice. The last aim focuses on understanding the role of TMEM135 in mitochondrial dynamics, thermogenesis, adiposity, and metabolic homeostasis using mice with adipose-specific knockout of TMEM135. Overall, this project has the potential to identify a novel organelle interaction regulating mitochondrial division, characterized by recruitment of peroxisomes to mitochondria, perhaps leading to new potential targets for therapeutic activation of BAT.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY/ABSTRACT Neuroinflammation plays a central role in the pathogenesis of Alzheimer’s disease (AD) and links strongly with AD’s neuropathological hallmarks, including amyloid plaques and neurofibrillary tangles. While there have been tremendous strides over the last decade in the development of novel cerebrospinal fluid, blood-based assays, positron emission tomography (PET), and magnetic resonance imaging (MRI) techniques, there still is a desperate need for a clinical imaging modality that can reliably image and quantify neuroinflammation in the aging brain and facilitate the identification of the key pathological players in different stages of AD. To address this unmet need and response to FOA “PAR-21-038”, we propose a new research direction to develop and establish a brand-new diffusion MRI (dMRI) technique, Diffusion Dictionary Imaging (DDI), as a safe, specific endogenous, and economical solution for the neuroinflammation imaging in AD. By tracking the random walk of water molecules using FDA-approved diffusion MRI sequence and compressed sensing technique, DDI measures and specifically extracts the microstructural changes associated with the activation and infiltration of the microglia and astrocyte in AD. Moreover, the previously collected dMRI data can be retrospectively analyzed with this new technique. Therefore, we are in a unique position to cost-effectively develop DDI by leveraging the longitudinal data-rich studies from the Knight Alzheimer’s Disease Research Center (ADRC) at Washington University (>800 Sporadic AD participants) and the multi-site international Dominantly Inherited Alzheimer Network (DIAN) observational study (>500 autosomal-dominant AD [ADAD] participants). In this project, we will develop the DDI technique and evaluate its capability of imaging neuroinflammation in the postmortem AD brains (Aim 1). We will examine the associations between the DDI neuroinflammation index and CSF and plasma inflammation biomarkers (YKL-40, sTREM2, and GFAP) in the Knight ADRC and DIAN cohorts (Aim 2). We will also cross-sectionally and longitudinally quantify the neuroinflammation using DDI in the Knight ADRC and DIAN cohorts and examine its connections with amyloid deposition and tauopathy measured by PET tracers and clinical outcomes (cognitive and progression) (Aim 3). The successful development of a quantitative endogenous DDI imaging biomarker of neuroinflammation will represent an important technological leap forward. The integration of DDI neuroinflammation biomarker into the DIAN and other clinical trial studies will provide clinically feasible neuroimaging surrogates that can significantly improve our understanding of the role of neuroinflammation in AD pathogenesis.

NIH Research Projects · FY 2026 · 2022-04

Project Summary The overall goal of this research is to better understand the how the kappa opioid receptor (KOR) system modulates cold hypersensitivity with the ultimate goal of uncovering a novel therapeutic target. Cold pain affects a number of diverse groups of patients and goes largely untreated. Neuropathic pain with cold allodynia is estimated to affect 15% to 50% of neuropathic pain patients. For many patients, cold pain is often a side effect that becomes a chronic debilitating condition. Patients undergoing chemotherapy using platinum-based drugs report increased sensitivity and pain to cold stimuli. Furthermore, heightened sensitivity to cold is problematic in those diagnosed with multiple sclerosis. For others, cold pain is part of larger more complex pain condition. The most commonly reported medications used to treat neuropathic pain are non-steroidal anti-inflammatory drugs (NSAIDs), opioids and anti-epileptics, which do not relieve or treat heightened cold sensitivity, primarily because we do not have clear understanding of the mechanisms involved in the modulation of cold pain. This research focuses on better understanding the mechanism by which peripheral KORs modulate cold hypersensitivity and cold pain. The first aim of this proposal will use combined transgenic and pharmacological approaches to assess the necessity of peripheral KOR expression in the modulation of cold hypersensitivity and cold pain. The second aim will determine the mechanism by which KORs in dorsal root ganglion can modulate TRPA1 signaling, as well as determine the necessity of the involvement of TRPA1 channels in KOR mediated cold hypersensitivity. The third aim will test whether KORs in dorsal root ganglion also modulate TRPM8 signaling. In the final aim, will use human dorsal root ganglion tissue from organ donors to determine if the KOR-modulation of TRP function exists in humans. Together, these approaches will allow us to dissect the role of peripheral kappa opioids in cold hypersensitivity and cold pain. Understanding the mechanisms by which the KOR system modulates cold hypersensitivity and how this system may also be involved in modulation of cold pain will have major implications not only in our understanding of basic mechanisms of cold hypersensitivity, but may also open up alternative therapeutic targets to allow us to treat cold hypersensitivity and pain.

NIH Research Projects · FY 2025 · 2022-04

Abstract Arthritis-related conditions occur in over 1 in 5 adults, and the prevalence is increasing. Current approaches to modulate endogenous inflammatory mediators in rheumatoid arthritis (RA) predispose patients to significant adverse effects (AEs), such as infection, when anti-cytokine biologic drugs are delivered continually at fixed doses. As the severity of RA fluctuates over time, development of specific therapeutic strategies that can sense and respond to varying levels of endogenous inflammatory mediators by producing correspondingly appropriate levels of anti-cytokine drugs represents an attractive alternative approach that may mitigate AEs induced by continuous biologic administration. The goal of this project is to used genetically engineered stem cells to create bioartificial implants for biologic drug delivery as a therapy for RA. By combining principles of synthetic biology and tissue engineering, we will develop stem cells that respond to specific pro-inflammatory cytokines such as interleukin-1, interleukin-6, and tumor necrosis factor alpha by producing targeted anti-cytokine drugs in a feedback-controlled, self-regulating, and multiplexed manner. A primary focus of this study is to fine-tune these reprogrammed anti-inflammatory cells to enhance the sensitivity and specificity of cell-based drug delivery in response to low-level systemic inflammation. Synthetic gene circuits will also be introduced in these cells to allow for exogenously-controlled tunable and inducible safety switches that can temporarily or permanently disable anti-cytokine drug production. These engineered cells will be encapsulated in agarose-based implants that will be placed subcutaneously in mice induced with experimental RA, and the long-term safety and efficacy of these approaches will be assessed using clinical, histologic, molecular, and pain/behavior testing. The creation of such “designer” cells provides the possibility for long-term, feedback-controlled drug delivery for the treatment of chronic inflammatory diseases.

NIH Research Projects · FY 2026 · 2022-04

Project Summary/Abstract Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with limited treatment options and unclear pathogenesis. Extracellular matrix (ECM) deposition by proliferating fibroblast populations drives the progressive airflow limitation and hypoxia characteristic of IPF, mediated by the cytokine transforming growth factor beta (TGFB). Direct inhibition of TGFB leads to intolerable side effects, and there is increased interest in therapies that instead modulate its regulation. Microfibril-associated glycoproteins 1 and 2 (Magp1 and Magp2, or Magps) are fibroblast-produced proteins which are secreted to ECM, anchor to microfibrils, and bind TGFB to limit its signaling under basal conditions, but have no known role in fibrosis. Magp knockout (KO) mice exhibit increased TGFB signaling in various tissues and have immune cell alterations due to bone marrow TGFB effects. In a single cell RNASeq screen, the genes encoding these proteins were markedly upregulated in fibroblasts in fibrotic lungs, and combined KO of Magp1 and 2 led to more severe fibrosis after bleomycin injury in mice. This suggests that Magps act as inhibitors of fibrotic signaling and follow up studies in Magp1 and 2 individual KO mice show that deficiency of both proteins is required to produce a pro-fibrotic phenotype. Which fibroblasts populations produce Magps and how they limit fibrosis is unknown. We hypothesize that Magp expression is an adaptive response to lung injury and limits fibrosis by sequestering TGFB in ECM and will explore this central hypothesis through three specific aims: 1) Evaluate anti-fibrotic contributions of Magp1 and 2 in pulmonary fibrosis. 2) Evaluate Magp effects on TGFB signaling and immune responses in lung fibrosis. 3) Characterize Magp-producing fibroblast populations. Using combinations of KO and inducible/tissue-specific KO mice, we will determine the individual relationships of these proteins to fibrosis and whether they exhibit redundant antifibrotic functions in vivo. We will analyze alterations in TGFB signaling and immune cell infiltration in lung as possible underlying mechanisms of Magp protection from pulmonary fibrosis. Finally, we will employ our recently developed single nucleus RNASeq approach along with single nucleus assay for transposase-accessible chromatin (snATAC)-Seq to characterize the fibroblast populations that express Magp1 and 2 and evaluate how lung-wide transcriptomes and epigenomes change in the absence of Magps during fibrosis. This proposal will provide Dr. Koenitzer with the mentored training in single cell sequencing and bioinformatics, fibroblast and ECM biology, and lung histology/imaging needed to achieve his goal of independence as a physician-scientist. Under the oversight of an expert scientific advisory committee, He will realize career milestones, complete formal coursework, and develop lasting collaborations. Findings from this work will have implications beyond IPF in other interstitial lung diseases and enable Dr. Koenitzer to develop new approaches to the treatment of lung disease as an independent investigator.

NIH Research Projects · FY 2025 · 2022-04

Project Summary/Abstract The prenatal period is a sensitive and critical time for brain development characterized by waves of neurogenesis, neuronal migration, and formation of neural networks. In the first and second trimester, microglia are the dominant immune cells of the brain and participate in a variety of processes essential to brain development, including secreting neurotropic factors and engulfing apoptotic neural progenitor cells. Fetal microglia dysfunction can lead to aberrant cortical lamination, resulting in an increased risk of brain pathology. We have identified numerous ligand-receptor pairs involved in microglia-to-cortex and cortex-to-microglia signaling predicted to contribute to human fetal microglia function and fetal brain development. We observe concordant expression of these ligand-receptors pairs in cerebral organoids (COs) and induced pluripotent stem cell-derived microglia with our human fetal data. COs can model early human brain development, but current models lack the immune component of the brain. Our data suggest that induced pluripotent stem cell-derived microglia co- cultured with COs (oMGs) capture significant phenotypic characteristics of human fetal microglia. Thus, a systematic analysis of neural maturation following integration of microglia into COs is the first step in using this model system to interrogate the molecular mechanisms underlying how neuron-microglia interactions establish early brain circuitry. This proposal aims to use COs and oMGs to assess how brain environment signals and corresponding transcription factors contribute to fetal microglia behavior and microglial interaction with neurons in early fetal development. In Aim 1, completed in the K99 phase, I will test the hypothesis that integration of microglia into COs results in enhanced neural maturation. Additionally, I will test how perturbation of homeostatic brain environment signaling in microglia results in microglia dysfunction and altered neuronal subpopulations. In Aim 2, I will identify transcription factor networks underlying human and mouse microglia behavior throughout development, at homeostasis and after an inflammatory insult. The goal for Aim 2 is to uncover species- conserved mechanisms in microglia responses to inflammation for improved therapeutic targeting and murine modeling and to discover potential human-specific risk factors for disease. Additionally, I will test the hypothesis that microglial developmental transcriptional factors are re-wired following an inflammatory insult, leading to long- lasting changes in microglia behavior and disruption of brain circuitry. Studies in Aim 2 will be completed in the independent phase. My long-term goal is to elucidate the epigenetic mechanisms underlying neuronal-microglia communication in health and disease as an independent investigator. I have assembled a diverse group of highly skilled mentors who will ensure that I receive extensive training in neurodevelopment and assessment of neural circuits. My training will be further enhanced by the unique scientific environment of the UCSD research community, which is geared towards the development and usage of cutting-edge technology and analytic methods to assess cellular heterogeneity and dynamic cell-cell interactions in the brain.

NIH Research Projects · FY 2026 · 2022-04

Project Summary/Abstract Traumatic brain injury (TBI) is the most common cause of acquired neurological deficits in children. Mild TBI, or concussion, is highly prevalent in adolescence, and is linked with short- and long-term cognitive deficits, chiefly inattention. Attention-deficit hyperactivity disorder (ADHD) is similarly prevalent in children and represents a primary form of inattention that occurs even without a history of head injury. The phenotypic similarity between primary ADHD vs long-term symptoms of inattention subsequent to concussion is both a diagnostic dilemma and clinical window of opportunity into the mechanisms by which brain injury affects cognition. In line with this framework, the goal of this K23 award is to provide the applicant with training necessary to identify distinct signatures of brain activity in children with primary ADHD vs ADHD with a history of concussion, which will motivate mechanism-specific treatments for these inattention symptoms. The proposed research leverages the Adolescent Brain Cognitive Development study, which is the largest ever cohort of children with concussion paired with detailed cognitive and functional MRI data, to achieve this goal. It capitalizes on the the superb cognitive neuroscience and imaging expertise of the applicant's mentors and collaborators to achieve the applicant’s short-term goals of training in cognitive assessment (Dr. Barch), TBI research (Dr. Brody), ADHD research (Dr. Fair), task-based (stop signal, n-back) fMRI (Dr. Barch), resting state fMRI (Dr. Dosenbach), and multivariate imaging statistics (Dr. Thompson). The proposed training will facilitate the applicant’s long-term goal of becoming an independent physician-scientist in the field of cognitive neuroimaging. These training goals will be advanced through the proposed research. First, the applicant will use multivariate fMRI methods to detect distinct patterns of task-related brain activation (Aim 1a) and resting-state connectivity (1b) in children with ADHD and a history of concussion, compared to controls and either condition alone. Then the applicant will investigate the fMRI correlates of time since last concussion (Aim 2) using task-related and resting-state fMRI. Finally, the applicant will perform exploratory analysis on the effects of prescription stimulant medications on brain activity in these groups (Aim 3). Notably, the proposed methods and techniques can be extended to answer many other outstanding questions about the intersection between neurological and cognitive disorders in the ABCD and future imaging studies. With a research program that employs multiple converging techiques to interrogate neural signatures related to concussion and its cognitive sequelae, the applicant will continue to address research questions relevant to the NINDS throughout his independent career.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY / ABSTRACT This application proposes a five-year research career development program focused on determining how glycosylation epitopes expressed during metaplasia and cancer contribute to these clinically significant cellular transformations. The applicant, Jeffrey W. Brown, M.D., Ph.D., is an Instructor of Medicine in the Division of Gastroenterology at Washington University School of Medicine. Since completing gastroenterology fellowship, Dr. Brown had worked in the laboratory of Jason Mills, where he has discovered a novel cellular process that he calls cathartocytosis [Greek: cellular cleansing] which is used by cells to efficiently dedifferentiate in the processes of metaplasia and cancer. He has subsequently determined that cathartocytosis is annotated by the glycan 3’-Sulfo-LeA/C and mice null for galectins that preferentially bind this epitope either fail to perform cathartocytosis or fail to package sulfomucins into mature granules. As glycobiology is a new field for Dr. Brown, Stuart Kornfeld will serve as his primary mentor with continued support from Jason Mills (Co-Mentor). Together, the candidate will be uniquely positioned to acquire the knowledge and skill set necessary to develop an independent research program investigating how specific glycosylation epitopes modulate tissue transformation and differentiation states in metaplasia and cancer. The expression and secretion of sulfomucins is uniformly present in Barrett’s esophagus and esophageal cancer, is the defining feature of type III [high-risk] gastric intestinal metaplasia, and is currently the best biomarker for detecting high-grade dysplasia and cancer in pancreatic cystic lesions. Using a comprehensive approach involving cell lines, organoid culture, and murine models, the experiments proposed herein will determine the proteome carrying 3’-Sulfo-LeA/C, the cellular signaling and transcriptional profile regulating its synthesis and secretion, as well as the molecular mechanism by which specific galectins modulate cellular differentiation. Ultimately, with the mentorship provided by Stuart Kornfeld, Jason Mills, and the research advisory committee, the knowledge and technical skills derived from the proposed experiments, and completion of the outlined career development plan, Dr. Brown will be well-prepared to establish an independent research program and is expected to be highly-competitive for R01 funding.

NIH Research Projects · FY 2025 · 2022-04

Project Summary Vision arises from the combination of viewing and visual processing. How animals align viewing strategies with regional processing specializations to accomplish specific behavioral tasks is unclear. We recently discovered that mice use binocular vision to pursue and capture insects. Here, we follow up on this discovery to understand the retinal signals and downstream pathways that mediate binocular vision for predation (Aim 1) and control the gaze to keep targets within the binocular visual field (Aim 2). Mammalian binocular vision relies on the presence of ipsilaterally projecting retinal ganglion cells (RGCs). We recently reported that nine of the 40+ mouse RGC types have ipsilateral projections. Here, we test the hypothesis, that two ipsilaterally projecting RGC types, the sustained ON and sustained OFF alpha (sONα- and sOFFα-) RGCs, guide binocular predation. We have developed intersectional transgenic tools to selectively label, silence, and remove the ipsilaterally projecting sONα- and sOFFα-RGCs (~300 cells, ~0.6% of RGCs). The sONα- and sOFFα-RGCs show increased density and acuity (i.e., reduced receptive field size) in the ventrotemporal retina. In Aim 1, we will combine transgenic and immunohistochemical labeling and high-resolution imaging of whole retinas to understand the organization of the sONα- and sOFFα-RGC acute zone in the ventrotemporal retina. We will analyze how sONα- and sOFFα- RGCs encode local, global, and combined motion individually and as populations with targeted patch clamping and two-photon calcium imaging. Visual stimulus parameters will be based on analyses of our large 3D tracking dataset of mice hunting crickets. Next, we will assess the binocular processing of sONα- and sOFFα-RGC signals by specific neurons in the superior colliculus, the retinorecipient target mediating predation. Finally, we will measure the contributions of ipsilaterally projecting sONα- and sOFFα-RGCs to predation using selective silencing and removal and 3D behavior tracking. In Aim 2, we will test the hypothesis that ON-OFF direction- selective (DS-) RGCs control the gaze to keep prey within the binocular visual field during pursuit and capture. Combining two-photon calcium imaging and immunohistochemistry, we will analyze the topographic maps of DS-RGC direction preferences around the sONα- and sOFFα-RGC acute zone. We will use targeted patch clamp recordings and two-photon calcium imaging to understand how ON-OFF DS-RGCs encode local, global, and combined motion stimuli with ethologically relevant parameters and in vivo electrophysiology to analyze the transformation of their signals by specific neurons in the superior colliculus, which mediates gaze shifts. Finally, we will test the impact of removing direction selectivity from ON-OFF DS-RGCs on gaze control during predation. Our studies will reveal how two conserved RGC subclasses, their regional specializations in the retina (acute zones and topographic direction preference maps), and downstream pathways cooperate to align viewing strategies and visual processing for an essential survival behavior.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY Osteoarthritis (OA) is a debilitating disease of the synovial joints and is the leading cause of pain and disability worldwide. OA affects over 30 million Americans, but there are no disease modifying drugs. Obesity is a major risk factor for OA; however, it has been difficult to disentangle the role of low-level systemic inflammation from the effects of altered joint loading with increased body mass. While cartilage requires loading to maintain tissue homeostasis, previous studies have suggested that abnormal loading due to increased body mass may explain why individuals with obesity experience cartilage damage at elevated rates when compared to healthy weight individuals. However, many recent studies demonstrate that increased body mass alone does not explain OA damage in human and animal models, and that adipokines, inflammatory mediators from body fat, play an important role in OA pathogenesis. Additionally, previous studies from the Guilak lab highlight the synergistic importance of the mechanosensitive ion channels Piezo1 and Piezo2 in cartilage health and maintenance: obesity and an OA-relevant inflammatory mediator, interleukin 1 alpha (IL-1α), modulates and sensitizes Piezo channel function. As such, this proposal investigates if cytokine/adipokine signaling is the mechanism by which Piezo channels become hypersensitized to mechanical force, ultimately leading to increased chondrocyte death. The goal of this proposal is to directly investigate the interaction between obesity-associated inflammation and the mechanosensitivity of chondrocytes. This study on the role of altered mechanosensation in the pathogenesis of OA with obesity will lead to the ultimate goal of targeting these pathways to develop novel therapeutic approaches. Specific Aim 1 focuses on determining if obesity-associated inflammatory conditions alter Piezo expression in cartilage and if this increased expression translates to increased chondrocyte sensitivity to mechanical loads. This study hones in on key dysregulated adipokines with obesity: IL-1α, Leptin, tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6). Specific Aim 2 uses transgenic mice previously developed in the lab to investigate if the loss of chondrocyte-specific Piezo1 and/or Piezo2 ion channels protects against cartilage damage in an in vivo model of obesity and joint injury. Specifically, mice will be fed a high-fat diet (60% fat) and subjected to a destabilization of the medial meniscus (DMM) surgery, known to evoke post-traumatic OA. Together, both aims strategically develop atomic force microscopy (AFM) and calcium (Ca2+) imaging techniques with in vivo assessments of cartilage integrity to complement the use of genetically-modified mice in a model of HFD superimposed with DMM injury. The results from this study will help identify the mechanisms by which obesity affects cartilage health, ultimately leading to the development of OA disease modifying drugs, that may be more broadly applied to other tissues affected by altered mechanosensation of Piezo ion channels.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY Atherosclerotic cardiovascular disease, including coronary artery disease (CAD) remains a leading cause of morbidity and mortality worldwide. Although existing lipid-lowering therapies can now reduce low-density lipoprotein cholesterol to very low levels, there is a significant burden of residual risk, highlighting a need for novel non-lipid therapeutic targets. Genome-wide association studies (GWAS) hold great potential for identifying novel therapeutic targets but have largely failed to deliver on this promise in part because the causal genes underlying GWAS loci are unknown. Through a large-scale genetic association study of protein-altering variation we discovered a variant in the extracellular matrix gene SVEP1 that positively associated with coronary artery disease without any effect on plasma lipids. We have now found with Mendelian Randomization that SVEP1 protein levels are causally associated with CAD in humans. To conclusively demonstrate that SVEP1 is the causal gene in this risk locus, we generated complementary mouse models of Svep1 deficiency. Our preliminary data show that SVEP1 is made by vascular smooth muscle cells (VSMCs) in the atherosclerotic plaque and that depleting SVEP1 in the arterial wall decreases the development of atherosclerosis. Based on our preliminary data, we hypothesize that VSMC-derived SVEP1 promotes atherosclerosis by activating integrin and Notch signaling to influence the behavior and fate of VSMCs in a cell- autonomous manner. In this proposal, we propose the following series of experiments to test this hypothesis: in Aim 1, we will define the cellular effects of depleting SVEP1 in the development of atherosclerosis; in Aim 2, we will define the molecular mechanisms by which SVEP1 influences VSMC phenotypes; and in Aim 3, we will determine if the pro-atherogenic and cellular effects of SVEP1 on VSMCs are dependent on binding to its partner integrin α9β1. Our investigative team has developed substantial preliminary data to support all proposed studies which are poised to reveal the mechanisms by which SVEP1 promotes atherosclerosis while providing new insights into the pathogenesis of CAD with the potential to reveal novel therapeutic approaches.

NIH Research Projects · FY 2026 · 2022-04

ABSTRACT Around 18,000 Americans suffer new spinal cord injuries (SCI) each year. Primary and secondary damages caused by SCI permanently impair sensory and motor functions, which require long-term therapeutic, rehabilitative, and psychological interventions. Thus, developing therapies to treat or reverse SCI is a pressing need in regenerative medicine. In contrast to mammals, teleost fish naturally regenerate functional neural tissue and reverse paralysis after complete spinal cord transection. Although innate spinal cord repair in zebrafish has fascinated scientists for decades, the contribution of glial cells to this elevated regenerative capacity is vastly understudied. Following SCI, adult zebrafish initiate a glial bridge that reconnects the severed spinal cord and supports functional neural repair. Pro-regenerative glial bridging distinguishes the zebrafish spinal cord and occurs without the detrimental outcomes of reactive gliosis elicited by the mammalian spinal cord. We propose that zebrafish glia orchestrate a series of transient, pro-regenerative responses that enable spinal cord repair. In this proposal, we will 1) determine the early injury responses that initiate glial bridging, 2) identify glial bridging mechanisms that are specific to zebrafish bridging glia and absent in mammalian glia, and 3) uncover the cellular and molecular mechanisms that direct glial bridge disassembly after regeneration is complete. This study will provide a mechanistic understanding of glial cell biology during zebrafish spinal cord regeneration, and will guide approaches for manipulating glial cells to promote spinal cord repair in mammals.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY The primary goal of this application is to determine the mechanisms by which pancreatic triglyceride lipase (PNLIP) variants increase the risk for chronic pancreatitis (CP) in humans. Proteotoxicity caused by misfolded mutant digestive enzymes has been recognized as a novel trypsin-independent mechanism for CP, referred to as misfolding CP. Recent studies indicate that the development of CP is often a process of discrete recurrent acute pancreatitis (RAP) driven by genetic risk factors. Restoring proteostasis in the presence of misfolded proteins is an attractive therapeutic strategy to stop RAP and prevent progression to end-stage disease in misfolding CP patients. Critical knowledge gaps remain in understanding how misfolded proteins contribute to pancreatitis onset and progression primarily stemming from the paucity of animal models that recapitulate misfolding CP in humans. We have developed a pre-clinical model of human PNLIP T221M disease. PNLIP has no known role in trypsinogen activation or trypsin inactivation. In vitro, PNLIP T221M misfolds and causes proteotoxicty by triggering unmitigated ER stress. Our preliminary data show that Pnlip T221M mice exhibit progressive pancreatic acinar atrophy, inflammatory cell infiltration, fibrosis, and apoptotic and necrotic cell death; the pathological changes occur as early as at 1 month and become severe at 3-4 month. Our working hypothesis is that proteins encoded by risk variants of PNLIP cause CP by gain-of-function proteotoxicity through protein misfolding. To test this hypothesis, first we will systematically characterize Pnlip T221 mice including CP-like phenotypes, secondary complications, and the susceptibility to pancreatic injury when being challenged with ethanol feeding. Second, we will methodically investigate early biological responses triggered by pancreatic expression of pathogenic PNLIP T221M in mice to uncover potential novel targets to halt or prevent disease onset and progression. Third, we will assess the prevalence of protein misfolding and proteotoxicity underlies the disease mechanism of PNLIP variants of uncertain significance (VUS) found in CP patients using cell cultures and another mouse model of Pnlip F300L. Completion of these studies will establish mutation-induced misfolding of digestive enzymes as a relevant disease mechanism in the pathogenesis of CP and will identify potential therapeutic targets to stop RAP and prevent CP.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY/ABSTRACT Lung cancer is the deadliest form of cancer, and more than 80% of lung cancers and lung cancer deaths are caused by cigarette smoking. Lung cancer screening with annual low‐dose computed tomography is recommended for long‐term current and former smokers, yet as few as 2% of 7.6 million eligible patients receive lung cancer screening. Most of these patients are current smokers, yet few receive effective tobacco treatment, with even larger care gaps among African American populations. This problem requires solutions at multiple levels, as uptake of lung cancer screening and tobacco treatment are driven by both physician orders and patient receipt of care. Novel, personalized efforts that target physicians and patients may boost uptake in lung cancer screening and tobacco treatment. We propose a multi-level intervention featuring a precision risk tool designed to stimulate guideline-concordant care by motivating behavior change and facilitating patient- centered discussions between primary care physicians and medically underserved patients at risk for lung cancer. This innovation is motivated by two key findings: 1) clinical and genetic factors inform precision risk for lung cancer and smoking cessation, and 2) high desire for personal genetic risk feedback signals its potential to activate behavior change. Building on important genomic advances, our team developed RiskProfile, a physician- and patient-facing tool that can incorporate genetic risk feedback to promote evidence-based care and cancer risk-reducing behaviors. The overarching goal of this study is to test the impact of RiskProfile, either with or without genetic information and in comparison to usual care, on uptake of lung cancer screening and tobacco treatment. We propose a 3-arm cluster randomized controlled trial of 75 physicians and 825 screen-eligible patients (11 per physician) from a diverse primary care setting. Physicians and patients will be randomized to usual care vs. RiskProfile-Clin (based on clinical factors) vs. RiskProfile-Gen (based on clinical and genetic factors) to evaluate the effect of precision risk interventions on lung cancer screening and tobacco treatment. In Aims 1 and 2, we will test the effect of RiskProfile on physician orders and patient completion of lung cancer screening and tobacco treatment. We predict that RiskProfile-Gen will outperform RiskProfile-Clin, and that both groups will outperform usual care. Primary outcomes will be ordering and completion of lung cancer screening among screen-eligible patients. Secondary outcomes will be ordering and receiving tobacco treatment among screen-eligible current smokers. In Aim 3, we will explore the impact of RiskProfile on potential mechanisms of behavior change (physician perceptions, patient cognitive or engagement factors, and physician-patient interactions) that may increase uptake of lung cancer screening and tobacco treatment. By targeting both physicians and patients and addressing both cancer screening and cessation care, this precision risk feedback tool has potential to drive down lung cancer incidence and mortality in underserved populations.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY Pulmonary hypertension (PH) is characterized by progressive increase of pulmonary vascular resistance and obliterative pulmonary vascular remodeling that result in right heart hypertrophy, failure, and premature death. The underlying mechanisms of vascular remodeling and obliterative vascular lesion formation remain unclear. Genetic mutations and variants were found in patients with idiopathic pulmonary arterial hypertension (PAH) and PAH with congenital heart disease. However, the mechanistic role of endothelial SOX17 in regulating pulmonary vascular remodeling in the pathogenesis of PH has not been reported. We hypothesis that endothelial SOX17 deficiency leading to activation of E2F1 signaling which contributes to endothelial hyperproliferation and anti-apoptosis in the pathogenesis of PH. We will 1) define the novel role of endothelial SOX17 in the pathogenesis of PH using multiple transgenic mouse and rat models. 2) delineate the molecular mechanisms downstream of endothelial SOX17 deficiency in mediating pulmonary vascular remodeling and PH and 3) explore the translational potential of targeting E2F1 signaling. Completing our proposed study will provide a novel therapeutic strategy for the effective treatment of PH in patients.

NIH Research Projects · FY 2025 · 2022-03

PROJECT ABSTRACT Globally, adolescents and young adults (AYA; ages 14-29) represent 30% of HIV incidence cases among persons of reproductive age, with ~75% occurring in Sub Saharan Africa (SSA). HIV incidence rises rapidly in SSA, as AYA leave school (often prematurely) and migrate for work and marriage. Older adolescents are more likely to experiment with health compromising behaviors that increase their vulnerability to HIV and sexually transmitted infections (STIs). Young people orphaned by AIDS [YPoAIDS]), 80% of which live in SSA represent a particularly vulnerable and unique population. Most YPoAIDS in SSA experience immense hardships, including higher rates of HIV risk behavior and odds of HIV infection. These adversities cumulatively disrupt the developmental milestones for YPoAIDS and can compromise their health and emotional wellbeing. Notably, our team has had great success with implementing a 6-year, three-armed RCT that tested a family-based economic intervention, Bridges (R01HD070727), among 1,383 primary school going adolescents in rural Uganda (10-14 years of age at enrollment) who lost one or both parents to AIDS. With over 90% retention rate over a 6-year period (2012-2018), our findings show efficacy of this contextually-driven intervention significantly improving sexual health, school retention and performance, and mental health. Yet, two critical policy and programming questions related to HIV prevention and engagement in care continuum remain unaddressed: 1) longer-term effectiveness of Bridges across YPoAIDS’s life course is currently unknown but critically important because of unique vulnerabilities during the transition into young adulthood; and 2) self-reports of sexual health are unreliable, hence the need to integrate biomarkers to provide the most precise results of these highly relevant (but currently unknown) sexual health outcomes among our participants. Thus, the specific aims of the Bridges- Round 2 study are: Aim 1. Examine the long-term impact of Bridges on: HIV prevalence (measured via participant’s HIV status) (Primary outcome); and b) Explore in secondary analyses the long-term impact of Bridges on key developmental and behavioral outcomes (e.g., mental health, alcohol and drug misuse); Aim 2. Elucidate the long-term effects of Bridges on potential mechanisms of change, including: a) economic stability, viral suppression (for ALHIV); PrEP use (for HIV negative adolescents), medical male circumcision (for boys); and b) young adult transitions; Aim 3: Qualitatively investigate participants’ experiences with Bridges that may have influenced engagement with the program, sexual risk-taking decisions, financial behaviors; experiences with developmental transitions; and perceptions on program sustainability; Aim 4: To assess the long-term costs and benefits of Bridges using formal economic evaluation. Our long-term goal is to translate knowledge into sustainable, theoretically-guided prevention and treatment efforts that promote the wellbeing of AYA affected by HIV/AIDS across the life course, in low-resource settings.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY/ABSTRACT This application describes our research into essential molecular pathways of the human pathogen, Mycobacterium tuberculosis (Mtb), including studies of transcription regulation and DNA repair. Infection with Mtb results in over 10 million new cases of tuberculosis and 1.5 million deaths annually, making it the deadliest infection in the world. In addition, this health crisis continues to be exacerbated by the emergence of drug- resistant strains, which demands the discovery of new antibiotic agents. In addition, we are deepening and broadening our biophysical work elucidating mechanisms of eukaryotic transcription initiation via both ensemble and single-molecule experiments coupled with kinetic modeling of the process in both yeast and humans. Transcription is responsible for changes in gene expression patterns during development or in adaptation to environmental conditions. The recruitment of RNA polymerase (RNAP) to particular genes at particular times is performed by sets of general and gene-specific transcription factors during transcription initiation. We are studying the essential, operator-independent, global transcription factors of Mycobacterium tuberculosis, CarD and RbpA. These factors act by modulating the rates of isomerization into and out of the open complex intermediate in initiation and, contrary to intuition, appear able to act as either activators or repressors without recognizing DNA sequence directly. We will answer critical questions in the field regarding the sequence- and sigma-factor (i.e., stress-response) dependence of these factors as well as their roles in post-initiation phases of transcription. We are also studying links between the transcription and DNA repair in Mtb. Mycobacteria lack classically conserved mismatch repair pathways (MMR) and possess repair factors not seen in E. coli. In addition, we have recently uncovered a novel oxidative switch that activates the Mtb nucleotide excision repair enzyme (NER), UvrD1. We are currently investigating the biophysical nature of this switch, alternative activation pathways, and the ability of UvrD1 to interact with RNAP during transcription-coupled NER. Of particular interest, and providing a link between our studies, is the shared RNAP-binding site used by both CarD and UvrD1. Lastly, we are continuing our investigations of the kinetic intermediates underlying pre-initiation-complex (PIC) dependent transcription initiation. Specifically, we are determining the mechanism of DNA bubble expansion during initial transcription in both yeast and humans. Our single-molecule magnetic-tweezers experiments will provide high-resolution views of the mechanism of PIC function. We are also following up on our recent discoveries of differences between the activities of yeast and human TFIIH (the general transcription factor required for promoter unwinding) that may underly the distinct usage of transcription-start sites in these organisms. As PIC function underlies gene expression, our unique approaches will provide important advances in the study of human biology.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY Despite advances in neonatal and neurosurgical care, post-hemorrhagic hydrocephalus remains among the most frequent, severe neurological complications of very preterm birth (gestational age at birth ≤32 weeks) and now represents the most common cause of pediatric hydrocephalus in North America. It also carries a heavy neurodevelopmental toll, with cognitive deficits and/or cerebral palsy diagnosed in greater than 75% of affected children. After the failure of medical approaches to impact its neurological sequelae, recent research has centered on optimizing neurosurgical treatment of post-hemorrhagic hydrocephalus, with a focus on mitigating ongoing injury due to progressive ventricular distension, a long-recognized risk factor for poor outcomes. The objectives of this proposal are to define the pathophysiological effects of post-hemorrhagic hydrocephalus on cerebral connectivity and neurological outcomes and, more specifically, to determine how ventricular volume modifies these relationships. Our Central Hypotheses are that 1) impaired structural and functional connectivity across key white matter tracts (e.g., corticospinal tracts, optic radiations, corpus callosum) and related functional networks (e.g., somatomotor, visual, default mode networks) are associated with neurological disability in post- hemorrhagic hydrocephalus, 2) ventricular distension contributes to post-hemorrhagic hydrocephalus-related connectivity deficits, and 3) these alterations in connectivity improve with neurosurgical ventricular decompression. Recent advances in MRI now enable characterization of functional and structural connectivity in the developing brain with unparalleled spatial and temporal resolution. Analysis of these data using the highly innovative diffusion basis spectrum imaging approach affords unique capabilities to characterize the complex neuropathological changes underlying these differences in cerebral connectivity. Here, our multidisciplinary team will employ these state-of-the-art MRI techniques in combination with detailed neurodevelopmental assessments to study a large cohort (N=180) that includes very preterm infants with and without post-hemorrhagic hydrocephalus prospectively recruited and followed longitudinally after discharge from the Neonatal Intensive Care Unit. In addition, infants with post-hemorrhagic hydrocephalus will undergo neuroimaging studies both before and after cerebrospinal fluid shunt surgery, characterizing the reversible effects on cerebral connectivity while also defining the role of ventricle size in its pathology. Application of these cutting-edge MRI acquisition and analysis approaches enables unprecedented characterization of the effects of post-hemorrhagic hydrocephalus on the developing brain. Further, we will extend these methods to delineate relationships between imaging measures and neurodevelopmental outcomes, improving our understanding of the modifiable effects of this devastating disease. Critically, these results will address long-standing, clinically important questions related to the care of infants with post-hemorrhagic hydrocephalus and inform development of innovative assessment tools to support clinical trials seeking to thwart the developmental disability observed in this high-risk population.

NIH Research Projects · FY 2026 · 2022-03

Parkinson disease (PD) is the second most common progressive neurodegenerative disorder characterized by tremendous clinical heterogeneity. PD with dementia is considered one of the Alzheimer disease related dementias. Dementia, afflicting up to 80% of PD patients and gait impairment cause significant morbidity and mortality, and tremendously amplify the socioeconomic burden as effective treatments are lacking. Both these impairments are refractory to dopaminergics suggesting involvement of other neurotransmitter systems or brain regions beyond nigrostriatal pathways. Most neuroimaging studies have focused on striatal dopaminergic deficits and fail to adequately explain gait and cognitive impairments in PD. The midbrain pedunculopontine nucleus (PPN) sends cholinergic input to the cerebellar vermis, which has much greater cholinergic innervation in humans than the cerebellar hemispheres. Degeneration of PPN, known to occur in PD could lead to cholinergic denervation of vermis. Vermis modulates gait and cognition, but little is known about its role in PD. Recent cerebellar resting-state functional connectivity (FC) analysis demonstrates altered vermal FC with sensorimotor and association cortices in PD and their respective gait and cognitive correlates. This proposed study utilizing a multimodal approach will extend the prior work and investigate whether longitudinal changes in vermal FC reflect decline in gait and cognition in PD and whether cholinergic denervation of the vermis, as measured by vesicular cholinergic transporter activity (VAChT) PET imaging, underlie these deficits in FC and behavior. The long term goal is to investigate whether vermal FC and cholinergic PET measures can serve as antecedent markers of dementia and falls in PD. The proposed study serves three major objectives. The first objective is to investigate cholinergic denervation of vermis and two important aspects of vermal FC in PD: its ability to objectively reflect progression of gait and cognitive impairment and its ties to underlying cholinergic pathology as measured by VAChT PET, the two key attributes of a reliable imaging biomarker. The second objective is to leverage the highly conducive and supportive environment, access to wealth of cross-sectional and longitudinal multimodal imaging data and comprehensive, sophisticated behavioral measures, and an excellent team of mentors to provide the candidate with training in a) PET analysis and b) conducting longitudinal studies with special emphasis on multivariate, longitudinal statistics to ensure a timely transition to independence. The final objective for the candidate is to continue to author manuscripts especially involving PET and generate preliminary data towards a R01 application. Although with independent research aims, this study is privileged to use the data collected as a part of large scale longitudinal study with established funding. The proposed study could have substantial clinical impact by elucidating pathophysiologic bases of dementia and falls in PD, identifying potential therapeutic targets and possible metrics of target engagement, stratifying at risk patients before these disabling symptoms progress irreversibly, potentially leading to individually tailored therapeutic options in the future.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY Toxoplasma gondii is an opportunistic pathogen that causes disease in immunocompromised patients and in infants due to congenital infections. Following a brief acute phase, the parasite differentiates into a semi- dormant form called the bradyzoite, which resides within long-lived tissue cysts that are primarily located in the central nervous system (CNS). Despite a vigorous immune response, tissue cysts are not eliminated and they pose a risk of reactivation when immunity wanes. It is estimated that ~2 billion people worldwide are chronically infected with T. gondii and hence at risk of reactivation should their immune function decline. In many regions of South America, infection can lead to severe and recurrent ocular disease in immunocompetent individuals. Chemotherapy is available for acute toxoplasmosis based on a combination of pyrimethamine and sulfadiazine. However, there are complications due to the toxicity of pyrimethamine and allergic reactions to sulfa drugs. Moreover, this treatment poses risks during pregnancy due to teratogenicity. Current treatment is effective in controlling acute infection, but has minimal effect on chronic tissue cyst stages and hence is not curative. Although a number of new compounds have been shown to inhibit T. gondii growth in vitro, most have little effect on the chronic stage. Hence, there is a major need for new efforts to identify compounds for treating chronic toxoplasmosis. The goal of this project is to identify late stage preclinical leads that show potent inhibition of parasite growth in vitro, eliminate chronic infection in vivo, and that possess appropriate ADME and safety profiles for advancement. We will develop potent and selective inhibitors of TgCDPK1, which is an unique and essential enzyme in T. gondii. In preliminary studies, we have identified several lead compounds that are both highly potent and selective for TgCDPK1 over mammalian kinases. We will design and synthesize new analogs to improve potency, selectivity, CNS penetration, bioavailability, and ADMET-PK properties of these compounds. Specific criteria for potency, selectivity and ADMET properties will be used to advance compounds to in vivo testing. We have developed new quantitative assays for monitoring inhibition of acute and chronic stages of infection and we will employ animal models for monitoring the efficacy of compounds against reactivated toxoplasmosis in the CNS. Successful achievement of these milestones will deliver lead compound(s) for future IND-enabling studies with the eventual goal of curing chronic toxoplasmosis.

NIH Research Projects · FY 2026 · 2022-02

Non-small cell lung cancer (NSCLC) is the leading cause of cancer related morbidity and mortality in the U.S. With the increasing use of computed tomography scans and lung cancer screening, tumors are increasingly being detected at stage I disease, conferring a greater than 70% likelihood of cure. Surgery has been the traditional treatment for stage I NSCLC. However, population-based studies demonstrate a rapid increase in stereotactic body radiation therapy (SBRT) over the past decade. Retrospective studies comparing surgery and SBRT, including several publications from our group, have been hampered by relatively small sample sizes, lack of patient reported outcomes, and sparse information on comorbidities and cancer-related outcomes. Consequently, a recent systematic review concluded, “there is a need to compare both treatments in large prospective trials”. Unfortunately, multiple attempted randomized controlled trials comparing surgery to SBRT have failed to accrue and closed prematurely due to specialty bias and perceived lack of equipoise. Furthermore, our previous work has demonstrated that individual patient characteristics are crucial in the choice of therapy. In the absence of prospective comparative studies, treatment allocation is largely directed by institutional experience, retrospective data, and provider opinion. The lack of evidence-based treatment allocation in patients with stage I lung cancer remains a critical unmet need. To address this fundamental gap in knowledge, we will perform a prospective, pragmatic, multi-center cohort study to compare the effectiveness of surgery and SBRT for stage I NSCLC. This pragmatic study design is ideally suited to provide actionable results. Aim 1: To compare the effectiveness of surgery versus stereotactic body radiation therapy (SBRT) on stakeholder selected short and long-term outcomes in patients with clinical stage I lung cancer. Aim 1a: To compare 3-year disease-free survival (DFS) between surgery and SBRT. We will compare long-term DFS using propensity score matched cohorts. We hypothesize that SBRT will lead to fewer and less severe treatment- related complications, while surgery will result in longer DFS. Aim 1b: To compare patient reported outcomes (PRO) between surgery and SBRT. We will compare short and long-term PRO in propensity-matched cohorts using the Patient Reported Outcomes Measurement Information System. We hypothesize that SBRT will result in better short-term PRO with equivalent long-term PRO. Aim 2: To develop and validate prediction models for treatment outcomes for an individual patient with stage I lung cancer. In prospectively assembled cohorts of patients undergoing surgery or SBRT for stage I lung cancer, we will predict long-term DFS and PRO for an individual patient using regression techniques. These models will be validated and presented as a web-based tool for patients with stage I NSCLC. Our proposal will create a benchmark for personalized treatment allocation in lung cancer and provide actionable results to improve care for Stage I NSCLC patients.

NIH Research Projects · FY 2026 · 2022-02

Regulation of Osteocyte Survival by Fibroblast Growth Factor Signaling Pathways Summary Osteocyte death, one of the hallmarks of skeletal aging, contributes to the age-related decline in bone strength and the increase in age-related fractures. In addition to aging, many other factors also lead to osteocyte death, including unloading, sex hormone deficiency, glucocorticoid excess, inflammation, and osteoarthritis. Although osteocytes function as master regulators of bone remodeling, the underlying molecular mechanisms that sustain osteocyte viability are poorly understood. Our studies aim to fill a gap in knowledge on endogenous factors that maintain osteocyte viability and bone quality in adult bone, as this is paramount to maintaining bone health. This proposal examines a novel role for Fibroblast Growth Factor Receptor (FGFR) signaling in the maintenance of osteocyte viability and skeletal homeostasis. In a recent publication, we identified a novel requirement for FGFR signaling for osteocyte survival. We showed that conditional knockout of FGFRs in mature osteoblasts and osteocytes led to osteocyte death in juvenile (3-week-old) mice and secondarily, increased bone mass as these mice aged. In a preliminary follow-up study, we temporally inactivated FGFRs in osteoblasts and osteocytes in adult (12-week-old) mice to bypass any effects on developing or actively growing bone. This also resulted in osteocyte death after several weeks and increased bone mass after several months. These observations form the basis of our hypothesis that in mature adult bone, FGFR signaling is required for maintaining osteocyte viability and bone homeostasis. Our proposed studies will address the mechanisms by which FGFR signaling maintains osteocyte viability and skeletal homeostasis in adult mice. Using lineage tracing and anabolic loading we will determine whether existing osteocytes vs newly formed osteocytes are sensitive to loss of FGFR signaling. In vivo analysis will identify the primary mode of cell death and determine whether remodeling of the osteocyte lacunocanalicular system is a cause or a consequence of loss of osteocyte viability. In vitro analysis of osteocyte cell lines will determine if FGFR signaling pathways are required cell-autonomously for osteocyte viability. Our preliminary data also suggests potential clinically relevant circumstances for either gain- or loss-of- function of FGFR signaling in bone. Two FDA approved FGFR inhibitors (Erdafitinib, Pemigatinib) have a median treatment period of 5 months for cancer. Our studies suggest that prolonged treatment with FGFR inhibitors could affect bone homeostasis. We will thus test the effects of Erdafitinib on osteocyte viability and biomechanical properties of bone in adult and aged mice. Finally, we will determine whether activation of FGFR signaling in mature osteoblasts and osteocytes is protective under conditions that promote osteocyte death. Completion of these studies will establish a role and identify mechanisms for FGFR signaling in the maintenance of osteocyte viability and bone homeostasis in adults, they will evaluate potential adverse effects of FGFR inhibition on bone, and will identify new genes that could be targeted to promote skeletal homeostasis.

NIH Research Projects · FY 2025 · 2022-02

Project Summary/Abstract Genetically identical cells in the same environment can have large individual differences in gene expression. This “single-cell variability” in gene expression diversifies cell types during development, contributes to chemotherapeutic resistance in cancer cells, and hinders the production of pure cell types in reprogramming protocols. Although single-cell variability is an important property of gene expression in development and disease, the sequence features of promoters and the epigenetic features of chromosomes that predict a gene’s single-cell variability are unknown. To address this gap, we propose to develop a high-throughput technology that measures the single-cell variability of different classes of promoters in diverse chromosomal environments. We propose to integrate transgenes into thousands of locations across the genome and measure the resulting single-cell variability of transgene expression from all locations in parallel. These data will allow us to leverage existing epigenomic maps to identify the features of chromosomal environments that amplify or dampen single-cell variability in gene expression. We propose to apply this technology to multiple cell types and with transgenes carrying diverse promoters. The resulting data will be analyzed with a framework separates the independent contributions of promoters and chromosomal environments to single-cell variability, and also quantifies any interactions between specific promoters and chromosomal environments that impact single-cell variability. Our goal in developing this technology is to understand the properties of the genome that control the single-cell variability of mRNA expression.