University Of Colorado Denver

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT This K23 proposal aims to develop and evaluate applications of artificial intelligence (AI) to the diagnostic investigation of infectious keratitis, a major cause of blindness worldwide. This will be accomplished through three specific aims: 1) Develop and evaluate an AI model to identify the etiology of culture-proven infectious keratitis from an existing database of clinical photographs; 2) Externally validate model performance in a real- world, population-based sample of corneal ulcers; and 3) Develop and evaluate an additional AI model for automated microscopic diagnosis of fungal keratitis. The AI model developed in SA#1 will be trained using a clinical photography database (the Culture Positive Ulcer Database) collated from several NIH funded clinical trials for infectious keratitis (SCUT, MUTT I & II, CLAIR, and MALIN) conducted over the past several decades as part of the international collaboration between the Francis I. Proctor Foundation and Aravind Eye Hospital in India. This model's performance will be compared against human experts on culture-proven cases of infectious keratitis. A second repository of imaging and clinical data from corneal ulcers (the MADURAI database) currently in development will be used to externally validate the AI model developed in SA#1 (by estimating its sensitivity and specificity in a real-world sample) and to train the AI model in SA#3. To accomplish these research goals, we have established an international collaboration between the Casey Eye Institute, the Proctor Foundation, and Aravind. This provides an unprecedented opportunity to leverage the expertise of my mentors at Casey in artificial intelligence and computer vision-enabled diagnosis of ophthalmic diseases, the expertise of the world-class faculty at Proctor in epidemiology, biostatistics, and infectious keratitis, and the unparalleled volume of infectious keratitis and infrastructure for data collection at Aravind. This collaboration will facilitate the development of carefully designed and validated AI models which will guide earlier directed antimicrobial therapy and improve visual outcomes in infectious keratitis. My primary career goals are to establish myself as an independent clinician scientist performing research at the interface of technological innovation and international public health. My MPH, medical training, and research experience have allowed me to develop a strong foundation in public health, the clinical and surgical management of corneal infections, and medical informatics. Over the past nine months of K12 support I have begun developing expertise in machine learning and data science, establishing a foundation which I will build upon during this K23 award period. The successful application of AI to health care problems requires a multidisciplinary approach involving clinicians, AI methodologists, informaticists, and public health experts. This K23 will allow me to build skills and expertise in each of these disciplines and become well positioned to lead this movement in the coming years.

NIH Research Projects · FY 2024 · 2022-07

ABSTRACT Klinefelter syndrome (KS) is a genetic condition affecting 1 in 600 males who have an additional X chromosome (47,XXY) associated with multisystem manifestations and increased mortality secondary to disorders of insulin resistance. One of the hallmark features of KS is primary testicular failure resulting in hypogonadism, and to date androgen treatment has been the only therapeutic intervention studied in these individuals. However, insulin resistance and abnormal metabolism have been observed in youth with KS prior to the onset of testicular hypogonadism. Furthermore, testosterone replacement does not ameliorate these cardiometabolic deficits that the majority of these individuals experience. The underlying molecular mechanisms for the high prevalence of insulin resistance and exercise intolerance observed in KS are unknown. This application proposes that the additional X chromosome leads to metabolic aberrations that can be targeted for therapeutic intervention. Peroxisome proliferator-activated receptor alpha (PPAR-α) is transcription factor that regulates the expression of genes controlling fatty acid metabolism, inflammation, and oxidative stress. Low PPAR-α activity is associated with cardiometabolic profiles similar to that observed in KS, including adiposity, dyslipidemia, and exercise intolerance. Our preliminary data have shown a metabolome and transcriptome consistent with insufficient activity of the PPAR-α complex in males with KS. The central hypothesis of this study is that lower PPAR-α activity contributes to the cardiometabolic phenotype in KS and that increasing PPAR-α activity via PPAR-α agonist treatment will result in upregulation of gene transcription that will improve cardiometabolic physiology. In this proof-of-concept trial, we will first compare expression of PPAR-α-regulated genes from whole blood and skeletal muscle in adolescent and young adult males with and without KS, as well as systemic fat oxidation during submaximal prolonged exercise and tissue-specific mitochondrial ß-oxidation. Resting energy expenditure, metabolite concentrations, and patient-centered outcomes will also be obtained and compared between groups. The KS cohort will then receive intervention with a PPAR- α agonist (fenofibrate) for one month, and all outcomes will be reassessed. This study will lay the foundation for future investigation of the first ever non-androgen treatment to improve insulin resistance and exercise intolerance in males with KS. The study aims support the mission and priority areas of the NIDDK and are expected to have direct clinical implications for individuals with KS.

NIH Research Projects · FY 2026 · 2022-07

PROJECT SUMMARY The long-term goal of this project is to understand how oligodendrocytes, the myelinating glia cell type of the vertebrate central nervous system, are formed during development. In the ventral spinal cord and dorsal forebrain, neural progenitors first produce neurons followed by oligodendrocyte precursor cells (OPCs), which migrate and divide to populate the neural tube. Subsequently, many OPCs stop dividing and differentiate as myelinating oligodendrocytes. Whereas we know a considerable amount about the mechanisms that promote oligodendrocyte differentiation and myelination, we know very little about mechanisms that regulate the very earliest steps in specifying progenitors for oligodendrocyte fate. Based on our in vivo, live imaging-based fate mapping studies, we discovered that different spinal cord and forebrain progenitor cells produce neurons and OPCs. We have now used single cell RNA-seq analyses to identify a transcriptional state of these pre-OPC progenitors, and have identified candidate cell-intrinsic programs and cell-extrinsic pathways that are conserved between nervous system regions and across vertebrate species. Using zebrafish and mice as model systems, this project will identify the molecular mechanisms that guide formation of OPCs from neural progenitors. Specific Aim 1 will test a hypothesis that the pioneer transcription factor Ascl1 establishes the epigenetic and transcriptional state of pre-OPCs. Aim 2 will investigate the mechanisms by which Notch signaling selects a subset of neural progenitors for a pre-OPC fate. Specific Aim 3 will test a hypothesis that the transcription factor Gsx2 helps specify a pre-OPC state and/or regulates a pre-OPC to OPC transition. The results of this project have the potential for important new insights into developmental myelination, the causes of myelin disease and potential strategies to restore myelin following disease and injury.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Sex differences are commonly reported in Lewy body dementia, although the reasons are unknown. Phenotypic differences can be described by differences in the pathology and can be associated with differences in genetic risk loci. We hypothesize that Alzheimer's disease-related genetic risk factors will be associated with higher dementia risk in women with Lewy body dementia, given the higher likelihood of Alzheimer's co-pathology in women. However, even for women with pure Lewy body pathology, Alzheimer's phenotype is more common than Lewy body dementia phenotype, indicating sex differences for clinicopathologic correlations. This may be due to differences in regional pathology burden for men and women. Specific regional Lewy body and Alzheimer's pathology burden have been associated with different symptoms; and we hypothesize that men will have more neocortical Lewy body burden and women will have more pathology burden in the medial temporal lobe given the more common Alzheimer's phenotype in women. As Alzheimer's disease is the most common misdiagnosis for patients with Lewy body dementia, we will also develop sex-specific models with clinical and genetic variables to clinically differentiate Lewy body and Alzheimer's pathology. These findings will improve our understanding of the etiopathogenesis of Lewy body dementia, assist with patient selection for future clinical trials in both Lewy body dementia and Alzheimer's disease, and provide targets for precision medicine in these neurodegenerative dementias. The candidate is an Assistant Project Scientist (a mentored position) at the University of California San Diego. She has an MD and a PhD with prior training in neurodegenerative disorders, neuropsychology, and neuroimaging. She has a history of productivity, having conducted translational and clinical research in movement disorders, recently focusing on sex differences in Parkinsonian disorders. She is committed to a research career in translational research and proposes a comprehensive five-year plan of research and training to acquire skills in 1) genetic and genomic analysis, 2) interpreting neuropathological data, 3) performing statistical analyses with big data. During the award period, Dr. Bayram will build collaborative relationships with experts in the field of genetics, neuropathology, and bioinformatics to support her work as an independent researcher. This award will also support Dr. Bayram's professional development including training for grantsmanship, leadership, and administrative skills. Dr. Bayram will meet her goals under the guidance of a mentoring team including Dr. Irene Litvan (primary mentor), a world-renowned expert in cognitive decline in Parkinsonian disorders, Dr. Sonja Scholz (co-mentor, neurologist-neurogeneticist at NINDS), Dr. Ali Torkamani (bioinformatics mentor), Dr. David Salmon (neuropsychology mentor), Dr. Dennis Dickson (advisor, pathologist), Dr. Owen Ross (advisor, geneticist) and Dr. Abraham Palmer (advisor, geneticist), all of whom are well-established and NIH-funded researchers.

NIH Research Projects · FY 2026 · 2022-07

Project Summary Craniofacial development is a complex morphogenetic process, disruptions in which result in highly prevalent human birth defects. Signaling through the platelet-derived growth factor receptor alpha (PDGFRa) plays a critical role in this process in humans and mice. Pdgfra mutant mouse models display midline facial clefting phenotypes. Phosphatidylinositol 3-kinase (PI3K) is the primary effector of PDGFRa signaling during skeletal development in the mouse, leading to activation of the kinase Akt. A previous phosphoproteomic screen identified Akt phosphorylation targets downstream of PI3K-mediated PDGFRa signaling in primary mouse embryonic palatal mesenchyme (MEPM) cells, revealing a significant enrichment for proteins that regulate alternative RNA splicing (AS), such as the RNA-binding protein (RBP) Srsf3. Ablation of Srsf3 in the mouse neural crest lineage leads to facial clefting due to defective cranial neural crest cell proliferation and survival. Further, Srsf3 regulates the AS of transcripts encoding protein kinases in the mouse facial process mesenchyme to regulate PDGFRa-dependent intracellular signaling. These findings have shifted the paradigm on how RTKs regulate gene expression and have identified post-translational modification of RBPs involved in AS downstream of PDGFRa signaling as a critical mechanism contributing to craniofacial development. The goal of this proposal is to determine the molecular mechanisms by which Srsf3 activity is regulated to generate protein isoforms necessary for midface development. First, to identify proteins that differentially interact with Srsf3 depending on its phosphorylation in response to PDGFRa signaling, Srsf3-interacting proteins will be immunoprecipitated from MEPM cells with or without PDGF-AA treatment and identified by mass spectrometry. Separately, Srsf3-RNA interactions will be purified and sequenced in response to PDGF-AA ligand stimulation through enhanced crosslinking and immunoprecipitation analysis to identify which transcripts are directly bound by Srsf3 and to determine if the extent of RNA binding and/or sequence specificity of these interactions changes upon Srsf3 phosphorylation. Second, craniofacial phenotypes will be assessed in Srsf3 phosphomutant knock-in embryos and RNA-seq analysis will be performed to identify AS targets of Srsf3 that depend on Srsf3 phosphorylation. The relationship between PDGFRa and Srsf3 will be dissected using this allele in genetic epistasis experiments. Finally, craniofacial phenotypes will be assessed in Srsf3 ectoderm- specific conditional knock-out embryos. The AS targets of Srsf3 will be identified in the facial mesenchyme and overlying ectoderm at the onset of craniofacial phenotypes through RNA-seq analysis. The splicing programs regulated by Srsf3 in each compartment will be correlated with biological processes active during craniofacial development. This project will delineate a complete pathway from PDGFRa at the cell surface to Srsf3 and its target transcripts in the nucleus. These studies will provide significant insight into what are likely broadly- applicable mechanisms underlying gene expression regulation during mammalian craniofacial development.

NIH Research Projects · FY 2026 · 2022-07

Project Summary: Mis-regulation of G protein-coupled receptor (GPCR) trafficking and signaling is implicated in causing several diseases and the development of drug tolerance, having a major impact on human health. GPCRs evolved to be the most important means for communication between cells and tissues in higher organisms. They are responsive to a wide range of stimuli including light, odorants, peptides, neurotransmitters, and hormones, making GPCRs critical players in regulating human physiology. Owing to their importance, they are the targets for a third of all FDA-approved drugs. For signaling to be temporally regulated, after agonist stimulation, GPCRs are desensitized. This desensitization occurs as a two-step process: first by phosphorylation, then by binding to proteins called -arrestins. -arrestin binding promotes acute desensitization by blocking access of G proteins to receptors. In addition, -arrestins act as adapters to proteins involved in clathrin-mediated endocytosis, facilitating internalization of the GPCR. Once internalized, the fate of a GPCR can differ dramatically, from being rapidly recycled back to the plasma membrane to being degraded. While classically GPCR signaling was thought to be confined to the plasma membrane, it is now appreciated that GPCRs can also signal from various intracellular compartments. Though our understanding of G protein-mediated signaling has matured over years of study, our understanding of how GPCRs are recognized as endocytic cargo remains limited. An important protein complex for this process is retromer, which sorts cargo at endosomes for recycling. A key component of retromer, vps26, is structurally similar to -arrestins, and is important for cargo recognition. I hypothesize that arrestin domain proteins are a privileged scaffold for recognition and trafficking of membrane proteins. As a result, understanding the molecular mechanisms that determine how GPCR--arrestins assemble and disassemble, and how they are trafficked in a cell, will have a profound impact on our understanding of signaling from GPCRs and the action of drugs. Using the 2AR together with V2R and NTSR1 as model receptors, I will (1) characterize how GPCR--arrestin complexes assemble and disassemble, and how this is affected by membrane lipids, GPCR phosphorylation, and the presence of other binding partners. I will also (2) identify protein interaction partners of GPCR--arrestin complexes in cells to understand which factors regulate the rapid or slow recycling behavior of these receptors. Finally, (3) I will characterize the engagement of a GPCR by retromer. These aims will be addressed using single-molecule fluorescence spectroscopy, state-of-the-art mass spectrometry, and in-cell photo-crosslinking. These aims will answer long-standing questions pertaining to arrestin function, and open new lines of investigation into regulation of GPCRs at endosomes. My Mentor, Dr. Kobilka, co-mentor Dr. von Zastrow and expert advisors in proteomics and protein-protein interactions (Drs. Hüttenhain, Krogan, Ting) and arrestin proteins (Dr. Benovic), will provide me with the training necessary to complete these aims and launch my independent research career.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT Type 2 Diabetes (T2D) has been increasing worldwide, in both adults and youth. Youth-onset T2D is strongly associated with obesity, characterized by rapid β-cell failure, early morbidity and mortality, and it almost universally presents in mid-puberty, a period of physiologic insulin resistance. However, our knowledge of the pathophysiology of youth-onset T2D is limited as it is primarily derived from cross-sectional studies, and longitudinal studies do not span the entire pubertal transition or only include youth with obesity. Such studies are not able to provide an understanding of the physiologic changes in glucose-insulin homeostasis during puberty, and of how prior metabolic health influences them. We also know of several risk factors for youth-onset T2D, but we do not know how they operate. These gaps prevent us from accurately predicting and preventing youth-onset T2D. The overarching goal of this proposal is to improve our understanding of metabolic health and dysregulation during puberty, and their determinants. To address this goal we propose to extend the longitudinal follow-up of the Healthy Start (HS) Cohort, a maternal-offspring community-based cohort study that enrolled 1418 pregnant women and conducted detailed characterization of mothers during pregnancy, of offspring through age ~7 years (R01DK076648), and ongoing at ages 8-10 years (through the Environmental Influences on Child Health Outcomes –ECHO- Consortium, UH3OD023248). This cohort is now transitioning through puberty. We will follow up 500 youth age 10-15 years throughout puberty to address specific aims: Aim 1: Describe glycemic trajectories [A1c, fasting and postload glucose, area under the glucose curve, time to peak during an oral glucose tolerance test-OGTT], and their metabolic correlates, during puberty; Aim 2: Explore the sequence of metabolic changes linking early life exposures to pubertal glucose-insulin homeostasis; Aim 3: Assess the importance of established genetic risk factors and characterize gene-environment interactions on pubertal glucose-insulin homeostasis. An improved understanding of the physiological changes in glucose-insulin homeostasis as youth transition through puberty will provide foundational knowledge to better predict future youth-onset T2D risk. An improved knowledge of the sequence of metabolic changes resulting from early life exposures and influencing adiposity and glucose-insulin metabolism, in the context of genetic susceptibility to T2D, will inform potential prevention approaches.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY An adequate placental transfer of lipids is critical for normal fetal growth and brain development, yet the mechanisms involved in placental lipid handling and transport are largely unknown. Many important pregnancy complications such as intrauterine growth restriction (IUGR) and maternal diabetes are associated with disturbances in fetal fat deposition that may contribute to adverse short-and long-term outcomes. Although mechanistic links between placental lipid handling and transport, fetal fat deposition and fetal brain development remain to be established, emerging evidence suggests that these pregnancy complications are associated with altered placental lipid metabolism. The syncytiotrophoblast, the transporting epithelium of the human placenta, mediates the transfer of lipids from the maternal to the fetal circulation. There is now compelling evidence that complex lipid forms are produced in large amounts by the syncytiotrophoblast and are potentially released to the fetus. In particular, primary human trophoblast cells rapidly take up fatty acids and incorporate them to phospholipids. De novo synthesis and remodeling of phospholipids generates a range of biologically active intermediates, including Phosphatidic Acid which regulates mTOR signaling in cancer cells. Phospholipid remodeling generates Lysophosphatidylcholine species containing docosahexaenoic acid (LPC-DHA) which is the predominant form in which DHA is transported across the blood brain barrier, mediated by MajorFacilitator Superfamily Domain Containing 2A (MFSD2a),an LPC-DHA transporter. We found high levels of LPC-DHA in placental tissue and that umbilical vein LPC-DHA levels are higher than maternal circulating concentrations, suggesting that DHA is delivered to the fetus as LPC-DHA. In addition, we discovered that MFSD2a is expressed in the human syncytiotrophoblast basal plasma membrane, consistent with the possibility that this transporter mediates transfer of LPC-DHA to the fetus. These observations provide the premise for our central hypothesis that trophoblast phospholipid synthesis and remodeling are highly active, produces phosphatidic acid (which modulates TOR signaling) and LPC-DHA for transport to the fetus, mediated by MFSD2a. Our hypothesis is supported by compelling preliminary data including evidence that phosphatidic acid regulates placental amino acid transport mediated through mTOR signaling and LPC-DHA is produced in placenta and released to the fetus. We will use human placental tissue, maternal and umbilical blood samples collected from normal and complicated pregnancies, siRNA gene targeting approaches in cultured primary human trophoblast cells incubated in a physiological mixture of 13C-stable isotope labelled fatty acids and trophoblast specific gene targeting in mice to address this hypothesis. These mechanistic placental studies are highly significant because we will use nove approaches to establish that phospholipid synthesis intermediates are important regulators of function and critical mediators of placental transfer of DHA, essential for normal brain development. l

NIH Research Projects · FY 2024 · 2022-07

Project Summary The sense of taste is mediated by multicellular taste buds that each house 50-100 rapidly renewing taste receptor cells (TRCs). TRCs are categorized into three main types: type I glial-like cells, type II cells that detect sweet, bitter or umami, and type III cells that detect sour. As TRCs renew, the three types are maintained in relatively stable proportions, allowing the sense of taste to remain stable over time. However, rapid turnover of taste cells makes the taste system prone to disruption by certain drugs and diseases. Cancer patients with metastatic renal cell carcinoma (mRCC) being treated with tyrosine kinase inhibitors (TKIs) often experience taste dysfunction, or dysgeusia. The primary targets of TKIs used to treat mRCC are the receptor tyrosine kinases (RTKs) VEGFR and PDGFRβ, which are not expressed in taste tissue. However, these TKIs also inhibit many off-target RTKs like Met, Ret, PDGFRα, and c-Kit. According to our single-cell RNA sequencing (scRNAseq) data, these RTKs are expressed in taste tissue in subsets of progenitors and differentiated taste cells. This suggests inhibition of these off-target RTKs may be the cause of dysgeusia and that some or all of these RTKs are necessary for proper TRC renewal and taste homeostasis. To test the role of off-target RTKs in taste homeostasis, I treated lingual organoids with the TKIs Axitinib, Cabozantinib and Sunitinib, which inhibit different combinations of off-target RTKs and frequently cause dysgeusia in patients. I found these drugs did not affect progenitor cell proliferation but instead decreased the expression of certain differentiated taste cell markers. Specifically, all three TKIs decreased expression of the sweet cell marker Tas1r2, and Tas1r2 was the only marker affected by all three drugs. Importantly, the only off-target RTK inhibited by all three drugs is c-Kit, which we find in our scRNAseq data to be most highly expressed in sweet-sensing type II TRCs. These data strongly implicate c-Kit in sweet cell homeostasis. Further analysis of our scRNAseq data reveals that c-Kit's ligand - stem cell factor (SCF), is expressed by type I and type III TRCs, raising the possibility of c-Kit mediated crosstalk within taste buds. These data in sum lead me to my hypothesis that c-Kit signaling, stimulated by crosstalk with other TRC types, is necessary for the differentiation and/or survival of sweet-sensing type II TRCs. To test this hypothesis, my first aim is to determine if c-Kit inhibition by TKI treatment leads to deficits in sweet taste in vivo by treating mice with Axitinib and performing behavioral, electrophysiological and cellular assays. My second aim is to determine if c-Kit is necessary for sweet cell differentiation or survival. I will use two genetic c-Kit knockout models under different Cre drivers to knock out c-Kit expression at different points in TRC differentiation. Lastly, I will investigate c-Kit mediated crosstalk by genetically knocking out SCF from within taste buds. Completion of this project will illuminate the role of the previously unstudied RTK c-Kit in taste homeostasis.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY/ABSTRACT African Americans (AA) and European Americans (EA) have a similar prevalence of gastro-esophageal reflux disease (GERD). Nonetheless, when compared to EA, AA show a lower incidence of esophagus damage, metaplasia, and esophageal adenocarcinoma. Population genetics and molecular studies have implicated specific genes for these differences in human tissue; however, a lack of racially diverse human esophagus models hinders further investigation into the mechanisms and potential treatment options. We developed an ancestrally diverse stem cell/organoid biobank of human esophagus and a high-content, image-based screening assay to interrogate bile-acid injury response. Results showed that AA esophageal cells responded significantly differently than EA-derived cells, mirroring tissue profiling and clinical findings. Furthermore, we have previously reported that a key enzyme, glutathione-transferase theta-2 (GSTT2), is responsible for inactivating reactive oxygen species, thus reducing DNA damage, and is highly expressed in the AA esophagus. Utilizing the ancestrally diverse stem cell model, we show key associations of GSTT2 low levels with higher injury, consistent with primary human tissue response to injury. However, a direct role of GSTT2 in this response and mechanism/drugs to maintain epithelial homeostasis and fitness to esophageal cells remains to be elucidated. Hypothesis: Esophageal tissue from African Americans respond differently to gastric acid/bile injury due to higher expression of detoxifying enzyme GSTT2, and compounds that can stabilize GSTT2 will protect cells against injury. The three specific aims to be investigated in this proposal will involve primary tissue and stem cell-derived in vitro cultures to validate the molecular profiles and differences in injury response between EA and AA cells at the single-cell level (Aim 1), with genetic manipulation of GSTT2 to determine direct mediation of protection against injury (Aim 2), and a high-throughput unbiased characterization of injury response coupled with a drug screen to determine compounds that will inhibit bile/acids injury (Aim 3). Dr. Ferrer-Torres’ primary research goals in the K99.R00 program is to develop high throughput techniques that will allow her to study ancestrally diverse populations and their response to injury. Therefore, the K99 phase has been planned to train in stem cell genetic modifications and high-content phenotypic-based drug discovery. For this mentored phase, Dr. Ferrer-Torres will work with Dr. Jason Spence and co-mentor by Dr. Jonathan Sexton. The mentored K99 program has been designed for Dr. Ferrer-Torres’ gain expertise in these areas. In addition, Dr. Jules Lin and Dr. Marcia Cruz-Correa will serve as advisory postdoctoral committee members and advisors for clinical immersions. This will be carried out utilizing the exceptional resources available at the University of Michigan. This will impulse Dr. Ferrer-Torres’ goals and help her establish her independent research program focusing on racial disparities in esophageal diseases in her R00 phase.

NIH Research Projects · FY 2026 · 2022-06

Project Summary Women are more susceptible than are men to stress-related mood and anxiety disorders, underscoring the importance of identifying strategies to promote stress resistance in women. Exercise affords broad benefits to mental health in both sexes, but whether the degree of stress protection and underlying mechanisms differ between sexes is unknown. We recently discovered that female rats are more responsive to the stress-buffering effects of exercise than males. It takes voluntary wheel running (WVR) half the time to enable protection against the depression- and anxiety-like effects of inescapable stress (IS) in female rats (3 wk) than it does in males (6 wk). Enhanced stress protection from exercise in females is an entirely unexplored resilience phenomenon. The goal of this proposal is to examine the sex- and circuit-specific determinants of this process across multiple units of analysis (molecular, cellular, circuits, sex, and behavior). VWR prevents the behavioral sequelae of IS by constraining activation of serotoninergic (5HT) neurons in the dorsal raphe nucleus (DRN) during IS, but the mechanism by which exercise constrains DRN 5HT activity is unknown. Our preliminary data provide strong evidence that stress resistance in both sexes arises from engagement of sensorimotor circuits (dorsolateral striatum; DLS) responsible for maintaining exercise. We have found that the DLS is positioned to inhibit DRN 5HT neurons through a direct GABAergic projection (DLS-DRN circuit), and 6 wk of VWR potentiates the activity of the DLS-DRN circuit during IS. Importantly, although the DLS is required to maintain VWR behavior in both sexes, the DLS governs VWR earlier in females (4 d) than in males (4 wk). Dopamine (DA) in the DLS contributes to DLS-dependent behavior and females are known to have heightened stimulus-evoked dopamine (DA) responses compared to males. This is likely the case with VWR, as just a few bouts of VWR activates D1 receptor-expressing neurons in the DLS of females, but not males. The rapid recruitment of the DLS during VWR in females could accelerate plasticity in the DLS-DRN circuit required for constraining stress-induced 5HT activity. Indeed, stress resistance from 3 wk of VWR in females depends on activity of the DLS-DRN circuit during IS. These data suggest that once exercise becomes governed by the DLS, the DLS-DRN circuit now responds actively to future adversity, thereby inhibiting the DRN & enabling stress resistance. We hypothesize that DLS neural ensembles link exercise to stress resistance and are particularly responsive to exercise in females, due to heightened DA responses to exercise in females compared to males. Intersectional genetic approaches that tag, record, and manipulate neural circuit activity during initial exercise and later stress will be used to 1) identify the role of the DLS in the development of exercise-induced stress resistance and accelerated stress resistance in females, 2) determine the role of the DLS in the expression of exercise-induced stress resistance in both sexes, and 3) identify the role of DA in driving accelerated stress resistance from VWR in females and in determining whether an appetitive, sensorimotor experience enables stress resistance.

NIH Research Projects · FY 2026 · 2022-06

Project Summary My long-term goal is to become a successfully funded, independent neurobiologist, conducting research aimed at improving our understanding of learning and memory. This goal will be accomplished by the implementation of a highly coordinated, thorough, and integrated career development plan including components of mentored guidance and training, an adept research environment, and a complementary research project. First, a career development plan comprised of a mentor, and co mentors, courses and conferences has been established to provide the applicant with the skills and guidance to transition into a position of independence. The mentor, Dr. Diego Restrepo, is a successfully funded researcher and leading expert in neuroscience and the co-mentor, Dr. Jamie Peters, is a recognized leader in neural circuits controlling learned behaviors, primarily those controlling drug seeking and fear in which the prefrontal cortex plays an essential role. They are well suited to oversee my training. The mentoring team will provide critical guidance and feedback on aspects of career development and technical components high density electrode recording with Neuropixels, data analysis, and olfactory behavior tasks. The mentoring committee includes members at different levels of their career ranging from young to senior faculty who will specifically assist the applicant in components directly related to career transition. In addition, courses in MATLAB, python and conferences on neuroscience are included to facilitate these components of the training program. Second, an adept research environment – the Department of Cell and Developmental Biology at the University of Colorado, along with associated resources, core facilities, and faculty members not directly involved in the applicant’s advisory committee – is established to provide assurance of the applicant’s success. Third, a research project has been proposed to facilitate my training in exciting new areas essential for my path to independence while simultaneously maintaining the key goals of studying neuronal signaling networks as related to learning and memory. The research tests the important hypothesis that altering local and long-range interneuron coupling and CaMKIIα-mediated plasticity mediate changes in PAC that result in an increase in odor elicited change in high gamma phase reference power and an increase in accuracy for decoding odorant identity during learning in the go-no go task. Aim 1 will evaluate Closed loop optogenetic stimulation of local PV or SST interneurons or long-range interneurons targeting hippocampus or mPFC in the peak or through of theta will cause coherence of oscillatory coupling causing the mice to increase decoding accuracy. Aim 2 will evaluate transient disruption of CaMKIIα expression in local SST interneurons or long-range interneurons targeting mPFC causes a change in the transmission of information from the CA1 to mPFC disrupting the ability of these mice to learn to discriminate odorants. Collectively, this comprehensive career development and research plan will ensure my successful transition from a dependent research position into one of well-prepared independence.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Emergence of the highly transmissible SARS-CoV-2 B.1.617.2 (Delta) variant and return to in-person learning is rapidly increasing the COVID-19 disease incidence and transmission rate in children. Therefore, it is critical to protect younger children. Toward this goal, ongoing COVID-19 vaccine clinical trials aim to reach those as young as 6 months old. Because pediatric trials involve fewer participants and children have a lower rate of symptomatic infection, pediatric vaccine efficacy will be initially based on in vitro immunological parameters­ virus neutralization and antibodies to the receptor-binding domain (RBD) or S1 fragment of the SARS-CoV-2 spike protein. However, whether these same metrics reflect an effective vaccine response in a developing immune system (<5 years old) remains to be determined. Immune responses to vaccines are influenced by age-associated physiological changes, particularly in the first 5 years of life when changes occur in B and T cell differentiation and effector functions, affinity maturation of B cell responses, and myeloid subpopulations and their cytokine production. Additionally, in very young children mucosal lgA rapidly reach adult levels, while serum lgA only reaches adult levels in adolescence. This difference may account for the disparate COVID-19 disease incidence, transmission, and severity in children. How evolving changes in pediatric mucosal and systemic immune ontogeny affect SARS-CoV-2 infection- and mRNA vaccination-elicited immune responses are incompletely understood. The overall obiective here is to define mucosal and systemic SARS-CoV-2 infection- and mRNA vaccine- elicited molecular and immune cellular responses in healthy pediatric maturing immune systems and in pathological B cell states (inborn or acquired). In a Pfizer-vaccinated adult cohort, compared to healthy adults, we have found that B cell depleted adult multiple sclerosis patients exhibited a significantly increased RBD-specific CD8 T cell response, despite negligible production of anti-RBD lgG. Interestingly, in children homeostatic and induced lgA levels are minimally affected by B cell depleting therapies. Our central hypothesis is that mRNA vaccination within the pediatric population augments mucosal (IFN and lgA) and CD8 T cellular immune parameters in the youngest children (<5yo) relative to older children (>5yo). We predict that such immune profile will i) correlate with vaccine- and infection-elicited responses, supporting their limited infection pathology (Aim 1); and ii) become enhanced in those children with B cell deficiencies (Aim 2). To test this hypothesis and its predictions, we will i) establish a prospective longitudinal cohort of SARS-CoV-2 infected/vaccinated healthy and B cell deficient children across age groups; and ii) apply transcriptomic, immune phenotypic, and antigen-specific humoral and cellular studies to compare SARS-CoV-2 vaccine- and infection-elicited molecular and cellular signatures in healthy children and those with inborn and acquired B cell defects. Resulting insights will define metrics of infection/vaccine immunity, constituting an initial step toward establishing correlates of protection in immunocompetent/B deficient children.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY Chronic kidney disease (CKD) is a predominantly age-related disorder and is associated with accelerated cognitive function decline. Despite epidemiological evidence linking mild cognitive impairment (MCI) and dementia with kidney function decline, we still have an incomplete understanding of the factors involved in their pathogenesis suggesting that there are other unidentified age-related causal risk factors that drive the development and progression of MCI, dementia, and kidney function decline. Recent data indicates that aging humans accumulate leukemia associated somatic mutations in hematopoietic stem cells. These mutations appear to provide a competitive growth advantage to the mutant cells, allowing progressive clonal expansion which has been defined as clonal hematopoiesis (CH) and it is characterized by worsening chronic systemic inflammation. CH is associated with several pathological conditions including cardiovascular disease, however, its association with cognitive and kidney endpoints has not been explored. In addition, the underlying mechanisms driving CH remain to be determined. Hence, our primary objective is to establish an association between CH with MCI, dementia, and kidney disease progression. Given the interplay between CH and aging related changes in the bone marrow (BM) fat microenvironment, a key secondary goal of this application is to determine the role of BM fat in the evolution of CH. We will also examine blood metabolite signatures associated with higher risk for clinically meaningful cognitive and CKD endpoints and higher BM fat content. Our preliminary data demonstrated a significant occurrence of CH in patients with CKD. Also, we have shown that BM adipocytes produce a local inflammatory signature including increased interleukin-6 that promotes CH. These findings provide a strong premise for our hypothesis that CH associates with MCI, dementia and CKD, and that BM fat plays a significant role in CH development. We will leverage the unique resources of the Systolic blood Pressure Interventional Trial (SPRINT), including longitudinal data on assessment of cognitive function and CKD, biospecimens, and DNA enabling assessment of CH. In a subset of 6,000 SPRINT participants aged 50 or older with normal kidney function at baseline, we will determine CH at baseline using our targeted sequencing panel and analytic pipeline for somatic mutation calling and assess whether CH identifies those at highest risk for cognitive and kidney function impairment. We propose 3 aims; Aim 1: To evaluate the prospective associations between CH and metabolomics with incident MCI and dementia events among SPRINT participants with normal kidney function at baseline. Aim 2: To evaluate the prospective associations between CH and metabolomics with kidney disease progression events among SPRINT participants with normal kidney function at baseline. Aim 3: To define a link between BM fat and clonal hematopoiesis in CKD. Understanding the disease related risk factors associated with CH and the related mechanisms may uncover new ways to reduce the burden of MCI, dementia and kidney disease progression.

NIH Research Projects · FY 2025 · 2022-06

Summary Pre-mRNA splicing is essential for gene expression in all eukaryotes and errors in splicing cause genetic disorders and many other diseases. A thorough understanding of the molecular mechanisms of pre-mRNA splicing has the potential to provide useful approaches for human disease therapy. The splicing of introns is carried out through two transesterification reactions catalyzed by the spliceosome, a large RNA/protein complex composed of five snRNPs (U1, U2, U4, U5, U6) and many non-snRNP related protein factors. The spliceosome undergoes dramatic changes in a splicing cycle, generating the E, A, Pre-B, B, Bact, B*, C, C*, P, and ILS complexes. Genetic, biochemical, and structural studies in the past four decades have generated tremendous information on the mechanism of splicing, but significant knowledge gaps remain. Our goal in the next 5 years is to use a combination of structural, biochemical, and genetic/genomics approaches to understand the mechanism and modulation of alternative 5’ ss recognition, the mechanism of exon definition, and the mechanism and function of transcription and splicing coupling. We envision that these projects will fill several significant knowledge gaps and advance our understanding of the mechanism of splicing.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY/ABSTRACT Exposure to violence and trauma in Latin America significantly contributes to migration to the U.S., bringing attention to the mental health consequences of forced migration at the U.S./Mexico border. Traumatic events occur at every stage of migration and negatively impact mental health. Evidence suggests that Latinx immigrants in the U.S. enjoy an initial health advantage that erodes over time spent in the country as immigrants experience discrimination and are less likely to seek health and mental health care. Also, this health advantage comes into question when analyzed by sub-groups. Insufficient evidence exists of the intersection of trauma exposure; individual characteristics; and mental health outcomes of depression, anxiety, and PTSD among recent Latinx immigrant adults. This K01 Award proposes a mixed methods design that incorporates latent class analysis methods to develop mental health phenotypes of recent Latinx immigrants based on the intersections of trauma, individual characteristics and mental health. This research will develop ethical approaches to studying trauma and mental health cross-sectionally and set the foundation for future prospective research of mental health phenotype evolution to inform intervention development and delivery. This research has three specific aims: (1) Develop initial mental health phenotypes in a sample of recent Latinx immigrant adults using latent class analysis methods and examine their demographic and trauma exposure correlates (trauma exposure, sex, age, gender, marital status, education, country of origin and ethnicity); (2) Provide an in-depth understanding of participant experiences of trauma exposure and mental health to illuminate findings in SA1 using qualitative methods; and (3) Validate the phenotypes and any additional constructs that emerge from qualitative data with a second survey sample of 313 recent Latinx immigrant adults at the U.S./Mexico border. My career development plan includes three Training Goals: (1) Develop comprehensive expertise in structural equation modeling and latent class analysis methods as well as growth curve modeling and latent transition analysis; (2) Gain extensive expertise in qualitative and mixed methods research at the intersection trauma exposure, individual characteristics, and mental health of Latinx immigrants with an emphasis on space and time.; (3) Develop expertise in ethical conduct of human research with a public health impact, that includes understanding gender differences in trauma exposure, as well as recruitment and prospective retention of vulnerable populations; and (4) Develop broad expertise in dissemination and translation of research with immigrant populations for policy and practitioner audiences. This K01 Award will support my long-term career goal of becoming an independent investigator conducting ethical longitudinal and intervention research on the mental health of recent Latinx immigrants in the U.S. The research plan will produce data for a multi-site R01 prospective study on the trajectories of trauma exposure and mental health of Latinx immigrants to the U.S.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY/ABSTRACT My career goal is to establish an independent aging research program focused on designing and evaluating high quality, technology-based behavioral health interventions to improve self-management and quality of life (QoL) of community-dwelling older adults, particularly older family care partners (FCPs). More than 17 million FCPs, including family, friends, and relatives, provide care for older adults with chronic illness in the US. High caregiving burden places FCPs at risk for physical function decline, psychological distress, and poor health status. Specifically, older FCPs are more vulnerable than younger FCPs because of greater physical and psychological burden, more age-related health care needs, and less emotional and social support. Physical activity (PA) has the potential to promote the health and well-being of older FCPs. Because caregiving duties limit FCPs to the home and preclude face-to-face interactions (more so during the COVID-19 pandemic), using digital health technologies is an effective way to engage older FCPs in home-based PA interventions. However, there is a gap in research for digital health PA interventions targeting older FCPs; I intend to fill this gap. This K23 award will provide me with the protected time to acquire advanced skills and knowledge in (1) design and implementation of clinical trials in older adults, (2) physical activity behavior change, (3) expansion of health informatics, and (4) professional development for a successful independent research career. The research environment at the University of Colorado, the interdisciplinary mentorship team I have assembled, and support from the College of Nursing provide an exceptional training opportunity. The overall objective of the proposed study is to rigorously design and test a tailored PA coaching program using digital health technologies, TPA4You, and conduct a pilot RCT to assess feasibility of TPA4You and its effects on health outcomes among older FCPs. Older FCPs of patients with heart failure (HF-FCPs) will be the focus of this study because HF-FCPs are an understudied population that is vulnerable to their own health-related issues. The specific aims of this proposal are to: (1) develop the TPA4You intervention by evaluating its technology features (n=15; Year 1), (2) conduct field-testing of TPA4You to optimize usability and acceptability (n=20; Year 2), and (3) conduct a pilot RCT to assess feasibility and preliminary effects of the TPA4You intervention (n=62; Years 3-5). For Aim 1, using scenario-based design methods, we will elicit HF-FCPs' perceptions about the integrated technology components in one-on-one interviews. For Aim 2, we will iteratively enhance TPA4You to meet HF-FCPs' PA needs and optimize usability and acceptability. For Aim 3, older HF-FCPs will be randomized to receive the TPA4You program or attention control for 12 weeks. This proposed study's novel TPA4You program has the potential to increase PA, and improve physical function, psychological outcomes, and QoL among older HF-FCPs. This proposal will provide the foundation for an R01 efficacy study of the TPA4You program in a larger, more diverse population of older HF-FCPs.

NIH Research Projects · FY 2026 · 2022-06

Project Summary Non-infectious uveitis is comprised of a heterogeneous group of clinically-defined diseases for which empiric therapy fails many patients. Molecular characterization of uveitis, on the other hand, would result in more precise classification of disease subtypes, while simultaneously providing pathophysiologic insight with therapeutic potential. This could spare patients unnecessary toxic side effects or loss of vision due to therapeutic inefficacy. Consequently, there is a critical need to better classify uveitis subtypes in order to discover more effective strategies for targeted immune suppression. The long-term goal is to develop a platform for molecular characterization of uveitis that will eventually facilitate precision medicine treatment strategies. The approach is to utilize validated experimental techniques to explore novel observations obtained from single cell RNA-Sequencing (scRNA-Seq), a high-definition gene expression analysis of ocular immune cells. Specifically, the central hypothesis for this proposal, that type 1 conventional dendritic cells (DC1s) promote the resolution of ocular inflammation, is based on primary observations of human ocular DC1s obtained via scRNA-Seq. The overall objectives of this proposal are to a) ascertain the role of DC1s in ocular inflammation, and b) to determine how patient-specific alterations in DC1s contribute to the chronicity of uveitis. Toward these ends, the following specific aims will be pursued: 1) Test the hypothesis that the ocular microenvironment promotes pro-resolution DC1s. 2) Test the hypothesis that DC1s promote the resolution of ocular inflammation. 3) Test the hypothesis that patient-specific DC1 gene expression signatures predict uveitis chronicity. In aim 1, the effect of the ocular microenvironment on DC1s will be tested by a) analyzing aqueous-exposed human DC1s in vitro and b) analyzing murine DC1s in 2 distinct models of ocular inflammation in vivo. In aim 2, the specific effects of DC1s on disease chronicity during ocular inflammation will be explored using mice deficient in DC1s. Finally, in aim 3, the transcriptional signatures that best predict chronic and acute uveitis will be validated and used to a) identify peripheral blood biomarkers and b) identify potential therapeutic targets. This contribution will be significant because it will improve our ability to define uveitis entities based on pathophysiologic mechanisms that can then be appropriately targeted by precision application of immune therapies. This proposal is innovative in the use of scRNA-Seq to generate testable hypotheses from in-depth analysis of patient samples. Ultimately, improved molecular characterization of uveitis based on identification and validation of differential gene expression provides a scaffold for future analyses that will facilitate a precision medicine approach to immune therapy.

NIH Research Projects · FY 2026 · 2022-05

Project Summary/Abstract Barrett's esophagus (BE), a metaplastic change of the esophageal lining associated with chronic gastroesophageal reflux disease, is the only known precursor to esophageal adenocarcinoma (EAC). EAC is one of the most rapidly increasing cancers in the United States, frequently presenting at an advanced stage and associated with a dismal 5-year survival rate. Endoscopic eradication therapy (EET) is the standard of care for patients with BE and high-grade dysplasia (HGD) or mucosal EAC. However, a central unresolved issue is whether BE patients with low-grade dysplasia (LGD) benefit from EET. The diagnosis of LGD is far more common than HGD and is associated with a lower risk of EAC, so it is unclear whether the costs and complications of EET are justified in this group of patients or whether they should simply continue with periodic surveillance endoscopy. The presence of clinical equipoise and the importance of this question indicates that a trial of endoscopic surveillance versus EET in this patient population is an urgent, unmet gap in our current knowledge regarding treatment of this common condition. We are uniquely positioned to address this significant gap in knowledge as we have assembled a multidisciplinary team with the requisite expertise in the conduct of clinical trials and biomarker research to ensure successful design and high-quality execution of the SURVENT trial (Surveillance versus Endoscopic Therapy for BE with LGD). This multicenter randomized controlled trial (n=530) will compare endoscopic surveillance and EET for the management of LGD using uniform inclusion criteria, design and endpoints. This trial will also include an observational cohort arm for those who decline randomization but are otherwise eligible (up to 150 subjects). Following our achievements during the U34 grant period, we propose the following aims for the U01: Specific Aim #1 will compare the two approaches using the primary endpoint of neoplastic progression rate (progression to HGD or mucosal or invasive EAC). Specific Aim #2 will compare patient-centered outcomes such as health-related quality of life between the two treatment groups. Specific Aim #3 will determine the utility of molecular (TissueCypher and p53 immunohistochemistry) and imaging (wide-area transepithelial sampling – WATS) biomarkers to improve risk-stratification in BE with LGD patients undergoing surveillance and EET. Biological samples will also be obtained at pre-specified time points to establish a biorepository for future translational research initiatives. The relevance of this work to the public health is high. BE is a common condition, affecting 2-3% of adult US population and LGD is seen in up to 40% of BE patients. This is a precursor for EAC and millions of dollars are spent yearly on the management of BE and EAC patients. The impact of our innovative study will include identifying the best patient-centered treatment approach for BE patients with LGD, which will inform the care of thousands of patients annually.

NIH Research Projects · FY 2026 · 2022-05

Project Summary Concussions are defined as a mild form of traumatic brain injury that result in acute neurological dysfunction. Recent work suggests post-concussion aerobic exercise at an intensity level below symptom exacerbation is safe. Yet, clinical benefits from existing randomized controlled trials indicate substantial room for improvement. Also, we currently have an incomplete understanding of the neurophysiology underlying changes in response to exercise treatment. Identifying the precise exercise dose (volume/intensity) required to elicit a therapeutic response following concussion will lead to enhanced and more precise post-concussion rehabilitation strategies. Our published and pilot data indicate light post-concussion exercise was associated with faster symptom resolution time and less severe symptoms, yet we relied on self-reported data and observational designs. Furthermore, we have identified that the optimal exercise volume to differentiate those with/without concussion symptoms after one month was >160 minutes/week, which is higher than standard exercise volumes prescribed (>100 minutes/week), and in line with existing recommendations for cardiovascular health (>150 minutes/week). Beyond this, given the positive effects of regular moderate exercise to reduce inflammation (e.g., interleukin 6) and restore cerebrovascular regulation, these physiological functions represent viable and feasible rehabilitation targets. Thus, using a prospective randomized clinical trial design, we aim to identify if high dose exercise >(150 minutes/week at an individualized intensity level), relative to standard-of-care, results in: faster/slower symptom resolution, altered physiological function, or reduced secondary sequalae. Our multidisciplinary investigative team has expertise investigating concussion, exercise physiology, fluid biomarkers, cerebrovascular physiology, and psychosocial outcomes. Thus, we will enroll, initially test, and randomize adolescents ages 13-18 years old ≤10 days post-concussion to high dose aerobic exercise or standard-of-care (symptom limited, self-guided physical activity), and reassess upon symptom resolution and 8-weeks post symptom resolution. We will obtain cerebrovascular function and serum biomarker data at each visit, and quantify exercise, symptoms, and secondary sequalae continuously. First, we aim to examine how the dose (intensity, duration, and frequency) of an aerobic exercise program initiated within 10 days of concussion affects time to symptom resolution, relative to standard-of-care, among adolescents. Second, we aim to assess the mechanistic relationship between aerobic exercise, biomarkers of neuroinflammation, and cerebrovascular function. Third, we aim to elucidate how high dose exercise after concussion affects persistent secondary sequalae development (anxiety, depression, kinesiophobia, peer relationships, academic concerns). By challenging the currently accepted, exercise recommendations for sport-related concussion, we will break new ground toward improving rehabilitation strategies.

NIH Research Projects · FY 2025 · 2022-05

PROJECT SUMMARY Cardiovascular disease (CVD) is the leading cause of death in the United States, with disparately greater impact on racial/ethnic minorities and lower-income individuals. Risk for CVD begins very early in life as adverse exposures during critical periods (fetal development, infancy, and reproductive years) shift the health trajectory toward overt disease. Thus, efforts to promote and maintain cardiovascular health (CVH) from pregnancy onward are crucial to reducing the intergenerational burden of CVD. In response to the Early Intervention to Promote Cardiovascular Health of Mothers and Children request for applications (ENRICH RFA), we propose to leverage two widely-available interventions to promote CVH in diverse, under-resourced pregnant women and offspring: Nurse Family Partnership (NFP) and the National Diabetes Prevention Program (NDPP). NFP is a home visiting program in which nurses support first-time, under-resourced mothers from pregnancy to 24 months postpartum to improve prenatal outcomes, child safety and developmental outcomes, and economic self-sufficiency. NFP is available at >260 organizations nationwide, including our Denver Health partners who have delivered it to >2400 urban, under-resourced women since 2000. While NFP addresses some putative causes of compromised maternal and child health, CVH has not been comprehensively addressed nor evaluated. Thus, we propose to enhance NFP with CVH-focused programming adapted from the NDPP, a yearlong, evidence-based lifestyle intervention that promotes healthy eating and activity to reduce weight, dysglycemia, and other CVD risks, which we have delivered to >1600 diverse, under-resourced adults at Denver Health since 2013, including >340 young women. By leveraging unique strengths of NFP (home delivery model, professional nursing support, impact on social determinants of health [SDOH]) with those of NDPP (evidence-based content for CVH promotion), a combined intervention (NFP-Heart) rigorously evaluated in the ENRICH program has great potential to improve CVH outcomes in under-resourced populations. Our Colorado Nurse Family Heart Trial will evaluate NFP-Heart versus usual care among 600 nulliparous women (and their offspring) recruited in early pregnancy from 10 clinics in a safety-net, urban healthcare system. NFP-Heart will promote intergenerational CVH with programming delivered from 20 weeks gestation to 24 months postpartum by home visiting nurses. Our specific aims are 1) develop the local NFP-Heart and common ENRICH protocols, and conduct pilot studies to confirm feasibility and acceptability; 2) evaluate the effect of NFP-Heart on maternal and offspring CVH metrics; 3) assess the degree to which SDOH modify the effect of the NFP-Heart intervention on maternal and offspring CVH outcomes; and 4) examine the implementation of the NFP-Heart intervention and potential for sustainability using a mixed- methods approach.

NIH Research Projects · FY 2025 · 2022-05

Abstract. Obesity induces a chronic systemic inflammatory state characterized by impaired adipokine signaling, increased pro-inflammatory cytokine expression, inflammatory cell activation, enhanced generation of oxidizing species and pathogenic shifts in metabolic intermediates and microbial profiles. This impacts pulmonary function and increases the incidence of asthma and its exacerbations that are resistant to conventional asthma therapies. Unsaturated fatty acid nitration products (NO2-FA), generated by metabolic and inflammatory reactions, can orchestrate diverse adaptive signaling responses. When administered as pure synthetic homologs, NO2-FA mediate post-translational protein modifications that modulate activities of multiple enzymes, receptors and transcription factors regulating metabolism and inflammation. Oral administration of synthetic NO2-FA 10-nitro- octadec-9-cis-enoic acid (termed NO2-OA or CXA-10) is a safe, novel pleiotropic drug candidate that is a synthetic homolog of an endogenous mediator. In murine models of metabolic syndrome, obesity-associated allergic airway disease and pulmonary inflammation affirms that CXA-10 induces anti-inflammatory responses and normalizes airway function. We will evaluate the promising pharmacology of this new drug class via Phase 2 evaluation of the therapeutic effects of CXA-10 in subjects with late onset obesity-associated asthma. We will a) define changes in pre bronchodilator FEV1, asthma control, and methacholine responsiveness following daily oral CXA-10 administration to obese subjects (BMI >30) having airway hyperreactivity, via a blinded, placebo- controlled, double cross-over study design and b) evaluate the impact of CXA-10 administration on study subject nasal and pulmonary airway cell gene expression, urine, plasma and bronchoalveolar lavage inflammatory biomarkers and gut-lung axis microbiome responses. These mechanistic studies will reveal how CXA-10 directs the electrophilic NO2-FA-sensitive genome and microbiome to modulate systemic and airway metabolic and inflammatory intermediates that contribute to the obese asthmatic phenotype. We hypothesize that nitro-fatty acid-induced signaling and metabolic responses will improve lung function, asthma control and alleviate obesity-related airway hyperreactivity. To test this hypothesis, Aim #1 evaluates the clinical responses of obesity-associated asthma patients to the orally-administered nitro-fatty acid, CXA-10 and Aim #2 identifies the downstream host and microbial gene expression and metabolic responses of subjects before and after oral CXA-10 administration. Current data encourages that, in the setting of obesity, CXA-10 will limit lung dysfunction, promote adaptive signaling responses and shift gut bacterial populations and metabolic intermediates so as to beneficially impact the gut-lung axis.

NIH Research Projects · FY 2025 · 2022-05

Project Summary/Abstract This application for a K08 Award includes training and research activities that support my transition to an independent mental health services researcher focused on improving services for parents with mental illness. I am a PhD-trained clinical social worker and tenure-track Assistant Professor at Portland State University, with a background in providing mental health services to parents and families. I am completing an Institutional K12 Award to study the mental health needs of parents in the neonatal intensive care unit (NICU). This K08 Award will build off my qualitative research and advance my research career by providing protected time, mentorship, and structured support to 1) learn implementation science methods, 2) increase my understanding of integrated mental health services, 3) obtain skills to conduct rigorous randomized control trials, including statistical methods necessary to analyze results, and 4) gain the skills necessary to become a leader in the field of parent mental health services research. My multidisciplinary mentorship team brings expertise in deployment-focused and participatory methods, health services research, implementation science, parent mental health research, randomized control trials, health economics, health informatics, rural health, and biostatistics. Parents of NICU-hospitalized preterm infants are at extremely high risk for mental health conditions with potentially devastating consequences. Yet, perinatal systems of care do not systematically support NICU parent mental health. The U.S. Preventive Services Task Force and state-level Maternal Mortality Review Committees have called upon perinatal systems of care to prioritize parent mental health. The objectives of this proposal are to develop and pilot test a telehealth-based mental health screening and engagement program that supports parents as they transition from the NICU to home. The program will use a stepped-care approach to screen parents for depression, anxiety, and PTSD; provide a brief behavioral intervention to those who screen as having at least a low risk of these conditions; and provide a warm hand-off to community mental health services for those at medium to high risk. We will achieve the study objectives by meeting 3 specific aims: 1) to use a RE-AIM framework to develop the program in collaboration with former NICU parents and NICU and primary care stakeholders; 2) to conduct an iterative case series to work out practical implementation details and logistics, refine the mental health screening and engagement program, and qualitatively assess purported mechanisms of action; and 3) to conduct a pilot randomized control trial to assess the program's acceptability and feasibility; to test its preliminary efficacy in targeting mental health self- efficacy and other potential mechanisms of action; and to better understand facilitators to adoption, implementation, and maintenance. The proposed activities will provide preliminary data and allow for the refinement of data collection procedures necessary for an R01 application to conduct a Type 1 hybrid effectiveness/implementation trial in multiple NICUs across the U.S.

NIH Research Projects · FY 2025 · 2022-05

Abstract The canonical functions attributed to telomeres in textbooks are to protect chromosome ends from degradation and fusion, both of which are confirmed drivers of genome instability and tumorigenesis. We have discovered two new and unforeseen roles for telomeres that are crucial for safeguarding the genome. First, we found that by interacting during meiotic prophase with the LINC complex (linker of nucleo- and cyto-skeleton), which spans the nuclear envelope, telomeres promote the nuclear envelope breakdown needed for spindle formation and the meiotic nuclear divisions. Remarkably, centromeres perform this function analogously in mitotic cell cycles, and indeed sporadic contacts between centromeres and LINC during meiotic prophase can rescue the loss of telomere-LINC contacts, indicating a surprising instance of telomere-centromere interchangeability. What features of telomeres and centromeres endow them with the capacity to control nuclear envelope breakdown and therefore cell cycle progression? Here we propose a series of experiments to answer this question. Second, we found that by providing a nuclear microdomain conducive to centromere assembly, telomeres rescue a surprising tendency of centromeres to become dismantled upon meiotic onset by the very factors (the meiotic endonuclease Spo11 and the meiosis-specific cohesin Rec8) that define meiosis. Indeed, we found that expression of Spo11 or Rec8 (which are normally meiosis-specific) in proliferating cells induces centromere dismantlement and chromosome missegregation. Here we propose to determine the mechanisms of Spo11- and Rec8-mediated centromere dismantlement, how telomeres promote the reassembly of dismantled centromeres, and whether these observations are relevant to the growing list of human cancers that mis-express meiotic proteins. These studies will open up new frontiers by defining at the molecular level unanticipated features of two key lynchpins of chromosome stability, telomeres and centromeres.

NIH Research Projects · FY 2026 · 2022-05

PROJECT SUMMARY Cortical subregions are often implicated in a variety of behavioral functions, but it is not well understood how these areas encode such diverse information. The anterior cingulate cortex (ACC) is necessary for emotional processing and social cognition, but how it encodes stimuli relevant to both processes is unknown. The goal of this proposal is to understand the functional heterogeneity of ACC circuits, and how this impacts encoding of diverse stimuli and cognitive function. In Aim 1, we will determine the functional heterogeneity of ACC inhibitory circuits during social and anxiety-like behaviors. To monitor interneuron activity, we will first inject an adeno- associated virus (AAV) that expresses GCaMP6f, a fluorescent Ca2+ indicator, in a Cre-dependent manner into the ACC of somatostatin (SST)- or parvalbumin (PV)-cre transgenic mice. Next, we will use the 3D-printed miniature miniscopes to perform in vivo single-cell resolution calcium imaging in either somatostatin or parvalbumin interneurons in the ACC to investigate their functional heterogeneity while mice performed tasks to assay anxiety-like behaviors, general sociability and social novelty. In Aim 2, we will determine whether specific populations of excitatory pyramidal cells (Pyr) in the ACC encode particular behavioral stimuli. First, to target specific populations of Pyr in the ACC, we will inject an AAV that expresses Cre recombinase and can retrogradely label cells into either the contralateral ACC, anterior thalamic nucleus, or the retrosplenial cortex. Next, we will inject an AAV that expresses GCaMP6 in a cre-dependent manner to label Pyr in the ACC and project to either the contralateral ACC, anterior thalamic nucleus or retrosplenial cortex. Three weeks later, we will use approaches described in Aim 1 to monitor the activity of specific populations of Pyr during particular behaviors. We will also determine the laminar sources of synaptic input to VIP interneurons in the ACC. To test this, we will first cross VIP-cre mice to a cre-dependent fluorescent reporter line. Next, we will inject an AAV expressing ChETA, a light-activated channel, into the contralateral ACC, anterior thalamic nucleus or retrosplenial cortex. Four weeks after viral injections, we will combine electrophysiological recordings in acute brain slices with optogenetic stimulation to characterize the laminar organization of specific projections unto VIP interneurons in the ACC. In addition to determining the connectivity probability of these inputs, we will correlate the cortical depth of the soma to morphological, and electrophysiological properties and molecular markers. While reductionist and simplified models of cortical networks have established a framework to understand their function, it is now evident that to understand the role of cortical networks in complex, adaptive behavior, optimized models will need to incorporate the functional heterogeneity of these circuits. Completion of these aims will dissect the functional heterogeneity of inhibitory circuits in the ACC.