University Of Colorado Denver

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY: With advancements in operative techniques and perioperative management, there is an increasing number of patients with Hypoplastic Left Heart Syndrome (HLHS) that are surviving into childhood and beyond. Due to the chronic pressure and volume load placed on the single systemic ventricle, these patients remain at constant risk for the development and progression of cardiac failure. Unfortunately, very little is known about how the failing HLHS heart differs from the failing adult biventricular heart, but the extrapolation of proven adult heart failure medications to the HLHS population has been unsuccessful, suggesting focused study is necessary to better understand the mechanisms underlying maladaptive remodeling and eventual heart failure in HLHS. Our preliminary data suggest glycosphingolipids (GSLs) play a role in modulating cardiomyocyte and immune cell function in HLHS, via both autocrine and paracrine signaling. Specifically, we show accumulation of LacCer in the myocardium of HLHS patients. To test the effect of LacCer on cardiac myocytes directly, we treated primary cardiomyocytes with liposomes carrying LacCer and determined that LacCer is sufficient to decrease mitochondrial OXPHOS, increase oxidative stress, and upregulate pro-inflammatory gene expression. Similarly, in vivo administration of LacCer induced right ventricular hypertrophy and decreased cardiac mitochondrial respiratory capacity. Further, we determined that PBMCs from HLHS patients display increased levels of LacCer, upregulated pro-inflammatory gene expression, and decreased mitochondrial OXPHOS. Similarly, treatment of healthy PBMCs with LacCer promotes decreased mitochondrial OXPHOS. Additionally, conditioned media from healthy PBMCs treated with LacCer is sufficient to promote mitochondrial dysfunction in primary cardiomyocytes, suggesting LacCer-induced crosstalk via paracrine mediated effects. Therefore, our central hypothesis is that aberrant glycosphingolipids drive maladaptive cardiac and immune cell responses and predispose HLHS patients to life-limiting complications including cardiac dysfunction. Aim 1 will investigate whether LacCer accumulation elicits a proinflammatory and metabolically dysfunctional phenotype in peripheral blood immune cells (PBMCs), and Aim 2 will elucidate the mechanisms by which GSL accumulation alters cardiac myocyte function. Our long-term goal is to ensure that HLHS patients benefit from rigorous pre-clinical studies in order to provide the necessary framework to identify novel therapies, influence clinical care, and improve outcomes. Successful completion of these aims will establish key mechanisms and modulators of heart failure progression in HLHS and will elucidate a potential therapeutic pathway to mitigate progressive cardiac dysfunction in this vulnerable population.

NIH Research Projects · FY 2025 · 2024-06

Project Summary This R03 proposal is to follow-up on recent findings from my SERCA K01 award, which focuses on elucidating the role of the type 1 diabetes (T1D) PTPN2 risk allele in loss of B cell anergy. Previously B cells bearing antigen receptors with high affinity for insulin were found only in the anergic B cell compartment of healthy individuals. Importantly, these cells leave this compartment in a proportion of first-degree relatives (FDRs), and in all autoantibody positive pre-diabetics and recent onset T1D individuals. Departure of these autoreactive anergic B cells in FDRs was shown to be associated with the high risk non-HLA allele, PTPN2 (rs1893217). PTPN2 has been previously shown to be a negative regulator of T cell signaling, but has not been studied in B cells. Recently we demonstrated that mice that lack Ptpn2 specifically in their B cells (Mb1Cre.Ptpn2fl/fl.C57BL/6) have a decrease in anergic B cells, similar to our findings in humans, an increase in autoimmune associated B cells (ABCs), increase in production of autoreactive antibodies, and a hyperresponsive phenotype. Despite these findings, these mice did not develop overt autoimmunity, likely driven by the fact these mice are on the autoimmune resistant genetic background, C57BL/6. Hence, in this study we aim to determine the effect of B cell specific deletion of Ptpn2 in the NOD mouse, which develops spontaneous diabetes by about 20 weeks of age. Specifically, we aim to study whether B cell deletion of Ptpn2 increases the rate and penetrance of diabetes incidence, the effect on frequency and activation status of insulin-reactive and non-reactive B cells, as well as its effect on B cell antigen presentation to T cells and the differentiation of pathogenic T cells. The potential impact of these studies will lie in understanding how risk alleles conspire to undermine maintenance of immune tolerance to autoantigens in T1D.

NIH Research Projects · FY 2026 · 2024-06

Project Summary Closed-loop VNS was recently approved by the FDA to restore upper limb mobility for patients with stroke. Moreover, preclinical and early clinical studies suggest that closed-loop VNS improves recovery from conditions such as spinal cord injury, traumatic brain injury, post-traumatic stress disorder and addiction. Despite the wide- ranging etiology of these conditions, the therapeutic model is similar; VNS is paired with a relevant rehabilitation protocol. The precise timing of stimulation is a key element to drive specific circuit plasticity and functional recovery. Yet, VNS activates widespread brain networks, raising the question of how closed-loop VNS can lead to circuit-specific alterations in plasticity and functional recovery. The goal of this proposal is to identify the mechanism by which closed-loop VNS drives circuit-specific plasticity during motor learning and stroke recovery. Our lab has shown, for the first time, that VNS elicits phasic cholinergic signaling from the basal forebrain (BF) and modulates inhibitory interneurons in motor cortex. Cholinergic signaling from BF is recognized to play a critical role in learning and plasticity and recovery from injury. The influence of cholinergic signaling on cortical plasticity is mediated through inhibitory interneurons, which gate synaptic plasticity within cortex. We hypothesize that closed-loop VNS drives recovery from stroke through cholinergic modulation of inhibitory interneurons to produce circuit-specific plasticity. The objectives of this proposal will be 1) to determine if closed-loop VNS drives synaptic plasticity on active circuits via cholinergic signaling; 2) to determine the influence of cholinergic signaling on inhibitory interneuron gating of synaptic plasticity and; 3) to explore the role of cholinergic activity and inhibitory interneuron gating of plasticity during stroke recovery. This data will provide a detailed mechanism by which closed-loop VNS can drive plasticity in the healthy and injured nervous system.

NIH Research Projects · FY 2026 · 2024-06

Project summary The meninges, located between the calvaria bones and brain, are composed of three layers; the pia, arach- noid, dura. Each meningeal layer contains specialized fibroblasts with unique expression profiles that facilitate specific functions. The meninges contain resident macrophages called BAMs (border associated macrophages), directly interact with bones of the skull, and contain extensive blood and lymphatic networks. The adult meninges provide physical protection, act as a selective barrier, function in neuro-immune surveillance, regulate cerebro- spinal fluid make-up, and are a site of CNS waste removal. During fetal development, meningeal fibroblasts are a key paracrine signaling source that regulates neurogenesis and cerebrovascular development via production of paracrine factors. Primary defects in meningeal fibroblasts cause congenital brain abnormalities in mice and humans. We have incomplete knowledge of the cellular and molecular mechanisms that control development of the meninges and its functions in fetal life. Our goal is to fill knowledge gaps on meninges formation and function and advance knowledge on how primary meninges defects may contribute to neurodevelopmental disorders. The transcription factor Foxc1 is required for meninges development and global Foxc1 mutants have been used for key discoveries of prenatal meningeal fibroblast function. To identify the cell-autonomous role of Foxc1 in fibroblast development and study meningeal fibroblast regulation of calvarial bone and local immune cell de- velopment, we generated conditional Foxc1 mouse mutants using Tbx18-CreERT that deletes Foxc1 expression in meningeal fibroblasts and mural cells but not other Foxc1-expressing cells like osteogenic cells. Dorsal fore- brain meningeal fibroblasts fail to specify into layer-specific subtypes in fetal Foxc1 conditional mutants, instead differentiating into an abnormal, myofibroblast-like cells. As a result of these primary defects in fibroblast devel- opment, we show resident meningeal macrophages (BAMs) do not develop properly and prenatal mutant mice have a severe calvarial bone growth defect. Aim 1 will identify how meningeal fibroblasts regulate BAM devel- opment, testing the function of meningeal-derived colony stimulating factor-1, and use a maternal infection model to identify prenatal functions of BAMs and fibroblasts in fetal neuroinflammatory response. Aim 2 will identify how signals from the meninges control calvarial bone growth over the brain, testing the prediction that meningeal derived ECM and secreted factors stimulate osteoblast cell migration. Aim 3 will employ single cell Multiome profiling and transcription factor-DNA binding analysis of Foxc1 and Foxc2 to identify the transcriptional mecha- nisms that drive meningeal fibroblasts to gain layer specific identities and the ability to produce paracrine factors that regulate brain, immune, vascular and bone development. Completion of experiments here will be impactful, uncovering new mechanisms into how the fetal meninges functions as a paracrine signaling center and generate potentially clinically relevant insights into how meninges defects contribute to neurodevelopmental disorders.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY. The mission of the NIH INCLUDE Project is to accelerate research that addresses the critical health and quality of life needs of individuals with Down syndrome (DS). A key aspect of this mission is to ensure that the many large and valuable multidimensional datasets that are being generated by INCLUDE-funded projects are available and amenable to a diverse community of researchers. Therefore, to increase diversity, equity, inclusion, and accessibility in the INCLUDE Project in particular and in the DS research community more broadly, we propose to develop an immersive summer course known as the Data Science for Diverse Scholars in Down Syndrome Research (DS3). This course will provide training in the generation, identification, and collection of high-content multidimensional datasets; their management, analysis, and visualization, including cloud-based analytics; and the development of professional skills required for the career advancement of diverse trainees. Led by a diverse teaching team, the DS3 will be developed along the following Specific Aims: Aim 1. To teach students how to perform FAIR research in the INCLUDE Data Hub and beyond. We will teach the FAIR principles (Findability, Accessibility, Interoperability, and Reusability) and demonstrate the FAIR use of datasets available in the INCLUDE Data Hub and other synergistic public repositories. We will tutor students about where to find datasets, how to frame answerable questions, considerations in using public data, and how to analyze these datasets in a cloud-based environment. Aim 2. To train students in the skills necessary for big data management, analysis, and visualization. Using short-read next-generation sequencing (NGS) data as a foundational data type, students will learn the basics of high-performance computing skills (e.g., Unix/Linux, cluster computing skills, file management and data analysis workflows). Students will be trained on the integration of -omics data types with clinical metadata, and also receive training in data visualization and biostatistical tools using Python and R Studio. Aim 3. To empower diverse trainees with professional skills necessary for career advancement. We will provide workshops on how to write a compelling Specific Aims page, how to develop clear presentations, and how to prepare an impactful curriculum vitae and NIH biosketch. Furthermore, we will hold networking events with prominent DS researchers and self-advocates who are invited as guest speakers. Aim 4. To develop a portfolio of DS3 training resources for dissemination at a global scale. We will develop a DS3 website connecting to public libraries of DS3 video tutorials, example analyses, presentations, slide decks, datasets, and code for sharing with the global community. Altogether, this training program will enhance diversity in the pool of young DS researchers, while also elevating data science literacy to accelerate clinical research that will benefit people with DS.

NIH Research Projects · FY 2025 · 2024-06

During animal development, signaling centers (a.k.a., ‘organizers’) play fundamental roles in the formation of organs, such as the CNS. Organizers consist of groups of cells that produce secreted factors (morphogens) that act on adjacent cells, often in the form of gradients, to induce distinct cell fates at different positions in the gradient. Dysfunction of organizers causes developmental disorders (e.g., agenesis of neural structures and limb defects). Hence, a key goal of developmental biology is to understand how organizers form and function. Several organizers have been implicated in CNS development (e.g., the anterior neural ridge organizer (ANR) and the isthmic organizer; IsO), but we do not understand how a population of neural progenitors is imparted with the ability to secrete specific morphogens and act as an organizer. The IsO is the most-studied CNS organizer as initial observations over a century ago defined a constriction (isthmus) forming in the neurectoderm at the junction of the midbrain and hindbrain primordia. The isthmus was later shown to contain a signaling center with the ability to produce Fgf8 and to control differentiation and positioning of adjacent neural structures, but we do not understand the cellular and molecular components required for IsO function. While the isthmic constriction appears to be positioned at the interface of the midbrain and hindbrain primordia – the IsO itself has been variably predicted to be derived from the midbrain or the hindbrain primordium, and we do not understand how the IsO is formed and positioned in the embryonic CNS. One key barrier to filling these knowledge gaps has been a lack of comprehensive molecular data for organizers during their formation in the CNS. A simple gene regulatory network acting at the isthmus was delineated decades ago, but it contains few gene. As a result, there is no clear molecular signature for the IsO, and we therefore cannot study its formation and function. Using scMultiome analysis – that enables simultaneous characterization of gene expression and epigenetic profiles of individual cells – we overcame this barrier and molecularly identified two cell populations that express known isthmic genes at zebrafish segmentation stages. One population co-expresses midbrain genes and one hindbrain genes, suggesting that they correspond to isthmic cells in the midbrain (IsMB) and hindbrain (IsHB), respectively. The IsHB also expresses fgf8 – indicating that it contains the IsO – but not all IsHB cells are fgf8-positive. At earlier stages (end of gastrulation), our analysis detects the IsMB, but the IsHB is not fully formed, and it is unclear if the IsO is associated with the IsMB or the IsHB at this stage. Our analyses represent the first molecular resolution of the isthmic region and includes earlier stages than prior work. These advances allow us to address key questions: What isthmic cell populations are required for IsO function? Does the IsO arise from the IsMB or the IsHB? We will answer these questions in the context of our hypothesis that the IsO consists of multiple essential cell types and derives from an early isthmic cell population (the IsMB) that possesses midbrain character.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Type 1 diabetes (T1D) results from the destruction of insulin producing β-cells upon immune infiltration and inflammation of the islets of Langerhans in the pancreas. Prior to T1D onset there is an asymptomatic phase of many years which presents a window for therapeutic intervention. However, there are limited means for detecting and tracking the development of insulitis in this ‘pre-symptomatic’ T1D window, to guide and monitor therapeutic interventions. Ultrasound imaging presents many advantages for clinical deployment, and gas- filled microbubble (MB) contrast agents are clinically approved, and used in detection and monitoring of a wide range of diseases. We previously used contrast-enhanced ultrasound to detect signatures of insulitis in pre- clinical models based on altered pancreas blood flow or inflammation-induced extravasation. While promising, a key limitation of pancreas imaging in general in T1D is that the islets of Langerhans represent only a few % of the pancreas volume. Any disease signatures will be ‘diluted’ by background signal, substantially reducing sensitivity. Thus high-resolution approaches to view the specific disease signatures in T1D are needed. Ultrasound localization microscopy (ULM) is a super-resolution microvascular imaging technique that provides microvessel-level spatial resolution noninvasively in vivo, thus potentially enabling the resolution of specific T1D disease associated changes in the pancreas. Our overall goal is to develop and apply novel ultrasound contrast agents in combination with ULM and signal processing to detect disease signatures associated with T1D, to provide image-based guidance and tracking of therapeutic interventions. We will conduct 3 specific aims: 1): Apply ultrasound localization microscopy to visualize vascular impairments in the pancreas. 2): Map the distribution of nanodrop accumulation and microvascular leakiness across the pancreas. 3): Apply molecularly targeted microbubbles and nanodrops to the inflamed and infiltrated islet microenvironment. Assessing inflammation and immune infiltration in the islet microenvironment using contrast-enhanced ultrasound imaging represents a fundamentally new paradigm for diagnosing and tracking T1D progression. The development and application of nanodrop agents and functionalized microbubbles, and use of super- resolution ULM techniques, represents further innovations which will have broader impact, aiding in diagnosing both the pace of T1D progression and the efficacy of any preventative treatment prior to clinical onset.

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY The goal of this proposal is to identify the mechanism(s) by which programmed death ligand 1 (PD-L1) reverse or intracellular signaling acts as an immune regulator in the liver during both acute infection, chronic infection and chronic disease. Our studies have outlined a major role for PD-L1 reverse signaling in the control of dendritic cell (DC) migration, DC activation and in regulating T cell cytokine production. These findings are consistent with PD-L1 acting to mitigate type 1 interferon (IFN) signaling events. Using a murine model where just three amino acids in the cytoplasmic domain of PD-L1 are mutated to alanine, to limit reverse signaling, we find increased fibrosis in a murine model of Metabolic dysfunction-associated steatohepatitis (MASH) and decreased liver pathology in an acute systemic listeria infection model. Therefore, in this proposal we aim to better understand the different functions of PD-L1 reverse signaling (ie, DC migration vs activation) and how they contribute to T cell programming in listeria infection and MASH. We also aim to better understand which PD-L1 expressing cells in the liver contribute to effector T cell responses and liver fibrosis in the absence of PD-L1 reverse signaling using a murine model of MASH. Finally, we aim to address the importance of PD-L1 reverse signaling in the liver in the setting of acute versus chronic infection of LCMV. Together these Aims will define how a widely targeted immune regulatory molecule, PD-L1, functions in the liver through intracellular signaling events not previously recognized or appreciated.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Group B Streptococcus (GBS), is a Gram-positive pathobiont that commonly colonizes the gastrointestinal and lower female reproductive tracts but can cause adverse health outcomes in neonates and vulnerable adult populations. GBS has also been increasingly associated with diabetic wound infections and is the third leading cause of death from skin and subcutaneous infection. It is estimated that over 500 million people have diabetes today with one of the most common complications being chronic wounds that fail to heal due to other diabetic complications and their high susceptibility to bacterial infection. Our lab recently performed dual RNA-sequencing on GBS infected wound tissues from diabetic mice (Db). Results showed significant increased transcription of many known GBS metal transport systems in Db wounds compared to culture controls including zinc (adcABC,adcAII,lmb), manganese (mtsABC), and an unknown metal transport system (nikABCDE). Preliminary results show that the putative GBS Nik transporter has high protein similarity and structure to the known E. coli nickel ABC-type transporter and is turned on under metal stress indicating it may play a role in GBS metal homeostasis. We also observed significant increased transcription and concentration of host calprotectin (CP), a neutrophil-derived metal chelator, in infected Db wounds compared to uninfected controls. Metal transport systems are key players in maintaining metal homeostasis of divalent cations, nutrients that are needed for all living organisms, while CP is a hallmark of inflammation and is involved in nutritional immunity where the host starves invading pathogens of nutrients necessary for survival. The role of the putative nickel transport system, NikABCDE, in GBS and during Db wound infection is unknown. Neutrophils also make a toxic byproduct of glycolysis called methylglyoxal (MGX) that can be broken down by a two-step pathway catalyzed by glyoxalase A and B. MGX is also increased in Db individuals and is a precursor to advanced glycation end products (AGEs) which further exacerbate Db complications. GBS contains homologous glyoxalase enzymes and preliminary data shows glyoxalase A contributes to GBS growth in the presence of MGX. This proposal seeks to characterize the putative nickel transporter in GBS and investigate the role of GBS MGX detoxification during Db wound infection. I hypothesize that the putative nickel transporter and MGX detoxification pathway are important for overcoming metal and MGX stress during infection. These hypotheses will be addressed with both in vitro biophysical and growth assays and in vivo models of Db wound infection in the following specific aims: (1) characterize the putative metal ATP binding cassette (ABC) transporter, NikABCDE, and (2) characterize the putative MGX detoxification system in GBS and its role during diabetic wound infection.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Alcohol-associated Liver Disease (ALD)-related mortality rates have increased 23% in the United States since 2019. While there remain concurrent increases in both the prevalence and mortality from alcohol misuse, patients have no therapeutic options outside from liver transplantation. Despite what is known about the pathogenesis of ALD, the mechanisms underlying the acceleration across the spectrum of liver disease remain poorly understood. It is known that patients with progressive ALD have progressive scarring of the liver and is the main predictor of patient outcomes. Excessive tissue scarring is characterized by highly cross-linked extracellular matrix (ECM) in response to injury. Hepatic stellate cells (HSC), the tissue resident fibroblast, play a key role in the initiation, progression, and resolution of liver fibrosis by transdifferentiating into activated myofibroblasts that secrete ECM in response to chronic injury, yet the mechanisms involved in HSC transdifferentiation are not well characterized. The goal of this proposal is to elucidate novel mechanisms of alcohol-induced fibrotic gene reprogramming in the liver and identify specific factors amplifying HSC activation in the diseased liver. While the uncontrolled regulation of Wnt/-catenin signaling is a hallmark of various hepatic pathologies including liver cancer, there are limited studies investigating alcohol exposure on canonical -catenin activation in specifically HSCs. Importantly, in canonical -catenin-dependent pathways, the association and release of the yes- associated protein-1 (YAP)/transcriptional coactivator (TAZ) (YAP/TAZ) from the -catenin destruction complex occurs in human embryonic kidney cells, yet it remains unknown if the convergence of these pathways occur in HSCs to drive their transdifferentiation in models of ALD. Our preliminary in vitro data show increased expression of -catenin and the transducers of Wnts, Frizzled (Fzd) receptors 1, 2, and 7, during spontaneous activation in primary mouse HSCs. Despite this knowledge, the exact role of Wnts driving the pathogenesis of fibrosis in models of ALD remain unclear. Our working hypothesis is that alcohol increases the expression of Wnts, leading to Wnt-dependent canonical -catenin activation, via YAP/TAZ, to promote hepatic stellate cell activation and production of ECM during the early pathogenesis of liver fibrosis in ALD. To test this, in Aim 1 we will determine if canonical Wnt ligands promote HSC transdifferentiation, in ex vivo models of alcohol exposure. Following treatment with ethanol and exogenous Wnt ligands, we will characterize HSC activation and fibrotic gene expression. We will then define if ethanol exposure impacts the sequestration and association of YAP/TAZ in the -catenin destruction complex to promote HSC transdifferentiation. In Aim 2, we will characterize the role of canonical Wnt signaling in HSCs using wild-type (C57BL/6J) and LRP6 knockout mice after ethanol exposure to measure primary outcomes related to hepatic fibrosis by gene and protein expression.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY/ABSTRACT This project addresses the NEI Audacious Goal Initiative (AGI) of restoring vision through regeneration of photoreceptor cells and their connections. Among degenerative diseases of the outer retina amenable to regenerative therapies, dry age-related macular degeneration (AMD) is a major target. AMD is the leading cause of irreversible blindness in developed countries, with the dry form accounting for nearly 90% of patients affected by this condition. Patients with end-stage dry-AMD present macular photoreceptor and retinal pigment epithelium (RPE) atrophy, and there is no available treatment that can restore their lost vision. To address this important unmet need we need to address two critical gaps: devising novel cell-based transplantation strategies to jointly regenerate photoreceptor and RPE cells, and identifying mechanisms supporting structural and functional integration of the transplanted cells. The goal of this project is to directly address these two critical gaps by establishing feasibility and mechanisms of structural and functional integration of a novel stem cell-derived 3D retinal transplant containing photoreceptors and RPE (3DNR/RPE) in a minipig model of end- stage macular atrophy. Our preliminary results showed survival of our 3DNR/RPE transplants within the subretinal space of our minipig model. This was accompanied by increased light response six weeks post- transplantation. These results strongly support our overarching hypothesis: that human induced pluripotent stem cells (hiPSC)-derived 3DNR/RPE transplants achieve structural and functional integration. In these studies, we will use an array of complementary multimodal in vivo ocular evaluation, ex vivo assessments, and non-invasive in vivo functional imagining to determine the extent of structural and functional integration of our hiPSC-derived 3DNR/RPE transplants. We will specifically test the following hypotheses: i) Co-transplanted photoreceptors and RPE re-establish physiological and functional interactions; ii) Transplanted photoreceptors synapse with both, co-transplanted and host bipolar cells, which in turn communicate with host amacrine and/or ganglion cells; iii) 3DNR/RPE transplants lead to increased light response from all photoreceptor cell types; and iv) 3DNR/RPE transplants lead to light-triggered response in bipolar and ganglion cells. This is a highly innovative strategy grounded in our ability to generate retinal tissue transplants derived from hiPSC that contain functional light-sensitive photoreceptors and RPE cells organized as in the native retina. Use of novel high-resolution technology for non-invasive functional assessment of transplant integration is an additional innovative aspect of our proposed studies. The potential impact of our studies is highly significant as it could lead to critical preclinical evidence supporting the feasibility of hiPSC-derived 3DNR/RPE transplants for cell- replacement therapy. In turn, this transplant strategy could become the first available solution for restoring vision in patients affected by end-stage dry-AMD.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY The Wind River Conference on Prokaryotic Biology (WRC) is held each year over four days in the Rockies. There are two features of this meeting that are unique, major strengths. First, the WRC is a small meeting that mingles investigators working on a wide variety of microbial questions. Second, interactions between trainees and senior investigators at the WRC take place on a level playing field, a situation one has to experience to appreciate. A critical goal of the WRC is to foster collaborations among and between junior and senior researchers who may only cross paths at a broad meeting such as the WRC. This enables junior scientists to build networks and identify opportunities for postdoctoral research and/or careers in government, industry or science writing, for instance. Our scientific program changes annually based on recent developments in the field, but each year we aim for strong representation in the areas of bacterial pathogenesis, physiology, and environmental microbiology. Many of our meeting participants, increasing so in recent years, direct or are supported by NIH-sponsored research programs studying diverse aspects of microbiology. We draw researchers from across the nation, with the aim of stimulating new ideas for collaborations on unexplored areas of scientific research. The active participation of principal investigators, including the keynote speakers, ensures the presence of an expert audience for the presentations given by the students and postdoctoral fellows. 1

NIH Research Projects · FY 2025 · 2024-05

Project Summary: RNA motifs can be assigned based on their sequence and/or structure, and are central to many biological processes. Examples of structural motifs include hairpins, internal loops, or bulges, which are known to serve several functions; similarly, examples of sequence motifs, can be those where a particular sequence is repeated, or conserved across different organisms/species, and also play important roles in biology. The Resendiz lab recently found that the palindromic RNA sequence 5'-GUAC- hybridizes with its complement, with an inherited disorder, when present in various duplex contexts, i.e., within GUAC repeats, double stranded chains of varying size, hairpin stem and loops, and internal loops. Preliminary data suggests that this `disorder' can be assigned to regions adopting a Z-type conformation. Importantly, this motif is found in various types of RNA that include viral RNA, micro-RNA, and transfer-RNA; thus the outcomes of this study will have broader implications in different biological systems. This work will establish the nature of the disorder found in the 5'-GUAC- RNA motif and explore its potential relevance in various biological constructs. The proposed work represents the first study that considers this sequence, as a motif, and that will establish the nature of the observed structural changes; which will be of potential importance, broadly. Structural aspects will be established using a combination of standard biophysical techniques that include circular dichroism, X-ray crystallography, NMR, electrophoresis, calorimetry, and organic synthesis of models that will probe various conformational spaces. The information will then be used to understand its impact on biologically relevant RNA. To this end, the structural impact of the 5'-GUAC sequence will be explored in two models for viral RNA [Human Rhino virus (HRV) and Severe Acute Respiratory Syndrome Coronavirus isotype-2 (SARS-CoV-2); and one for micro-RNA (miR-486). Understanding the elements that are necessary to exhibit the observed behavior, will allow researchers to treat/use this motif as an important structural element, within various constructs of RNA. The PI's group hypothesizes that this motif can become a potential druggable target, due to its unique structure and likelihood to interact with metabolites in a distinct manner; as well as a sequence with unique capabilities to interact with other biopolymers, i.e., RNA, DNA, or proteins. The proposed research has the potential to be of high impact, and of interest to various fields.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY The objective of this research is to understand the role and mechanism by which chemoattractant and integrin signaling plays in the bone marrow selection of B cells and in shaping the peripheral primary B cell repertoire and its degree of self-reactivity. The fact that 30% of the millions of B cells produced daily express a B cell receptor with high-avidity for self-antigen suggests we may not be healthy and live a full life in the absence of central B cell tolerance. Intriguingly, individuals affected by common autoimmune diseases display defects of central B cell tolerance manifesting with much higher numbers of bone marrow autoreactive B cells entering the peripheral tissue. However, these individuals also have defects in peripheral tolerance. A better understanding of how newly generated high-avidity autoreactive B cells are controlled by or escape tolerance can have profound implications for the development of therapies that may reduce the self-reactive burden of the circulating B cell population in individuals predisposed to autoimmunity. We have recently documented that the chemokine receptor CXCR4 is expressed at higher levels by autoreactive B cells undergoing central tolerance than non-self-reactive B cells licensed to leave the bone marrow. Moreover, in vivo treatment of mice with a CXCR4 antagonist, releases high-avidity autoreactive B cells from the bone marrow into the circulation from where they relocate into the spleen. The mechanisms by which CXCR4 signaling retains self-reactive B cells within the bone marrow is not clear. It is known that CXCR4 signaling can modulate the function of integrins, molecules that regulate cell adhesion to tissues. Chemokine receptors and integrins can also be regulated by PI3K signaling or can themselves promote the activity of PI3K, which is important for the positive selection of immature B cells out of the bone marrow. The goal of this grant is to define the contribution of CXCR4 and integrin receptors to central B cell tolerance and the impact of central B cell tolerance on peripheral tolerance and autoimmune responses. Experiments in Aim 1 will establish whether CXCR4 signaling controls the bone marrow retention of autoreactive B cells in human beings. Aim 2 will resolve the contribution of integrins in bone marrow B cell selection and the crosstalk between CXCR4, integrins and PI3K. Experiments in Aim 3 will test the hypothesis that the CXCR4-mediated central B cell tolerance process is necessary to reduce the chances of systemic autoantibody and autoimmune responses. Overall, these studies are significant because they generate a deeper mechanistic understanding of how B cell tolerance can be breached during the development of autoreactive B cells to raise the autoreactive capacity of the primary circulating B cell repertoire and the general risk for autoimmunity.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY. Individuals with Down syndrome (DS), the genetic condition caused by trisomy 21 (T21), display widespread auditory dysfunction, with current estimates indicating that ~80% of this population may have one or more forms of hearing loss. Despite the obvious importance of hearing for neurodevelopment, speech development, learning, cognition, social integration, inclusion in the workforce, and overall well-being, the mechanisms by which an extra copy of chromosome 21 causes hearing loss are unknown. Therefore, additional research in this area has potential for transformative discoveries that could improve multiple dimensions of development, health, and quality of life in this population. To fulfill this unmet need and transform the field, we have assembled a multi- disciplinary team of synergistic expertise to develop the following Specific Aims: 1. To complete deep phenotyping of peripheral auditory and vestibular function in DS. We will complete a deep characterization of auditory and vestibular function in a cohort of 300 participants via behavioral and physiologic assessments including air and bone conduction behavioral audiometry, otoacoustic emissions, standard tympanometry, wideband acoustic immittance, and auditory brainstem response testing. We will also administer a battery of vestibular function assessments and complete a detailed annotation of health histories, with an emphasis on co-occurring conditions and previous use of ototoxic medicines. 2. To identify dysregulated biological processes underlying hearing loss in DS. Paired to efforts in Aim 1, we will collect biospecimens, including blood samples and nasopharyngeal swabs, to complete both hypothesis- driven and hypotheses-generating analyses of underlying mechanisms, including analyses of the transcriptome, proteome, metabolome, neurodegeneration and neuroinflammation markers, and variations in the microbiome. These datasets will enable the identification of novel biosignatures associated with different etiologies of hearing loss and vestibular deficits. 3. To define cause-effect relationships between gene triplication and hearing loss in a mouse model. We will complete deep phenotyping of hearing loss in a mouse model of DS that displays this phenotype, including imaging studies to monitor for structural differences, wideband acoustic immittance measurements, distortion product otoacoustic emissions, tests of vestibular function, and measurements of otitis media. These studies will be complemented by assays not possible or very difficult in humans, such as single cell transcriptomics and cochlear endolymphatic potentials. Lastly, we will define the preclinical value of potential pharmacological interventions and narrow down the genomic region(s) driving hearing loss in mice. Funding of this proposal will enable a multidisciplinary team of experts to address a major knowledge gap in the field, with clear potential for transformative discoveries that would benefit those with DS.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY Perinatal mental health disorders are the most prevalent perinatal comorbidity and are associated with the primary cause of maternal mortality in the United States (US) – suicide. Diagnosis of a high-risk pregnancy and cesarean delivery (CD) are both associated with increased risk for perinatal mood, anxiety, and trauma symptoms (PMATS). There are over 1 million CDs annually in the US with one-third of these patients reporting PMATS. Directing prevention efforts toward high-risk pregnant patients requiring CD addresses risk for PMATS in a large patient population. There is a deficit in resources and access to mental health treatment for pregnant patients, with some treatments being cost prohibitive and requiring multiple sessions. Additionally, current approaches to addressing PMATS are reactive rather than preventive. There is evidence in the non-obstetric population that single-session cognitive behavioral therapy interventions targeting anxiety sensitivity (fear of fear) can prevent the development of anxiety and trauma symptoms when individuals are exposed to a trauma. We developed a low cost, 1-hour, single-session prevention intervention that included psychoeducation about anxiety sensitivity, coupled with a brief exposure to the operating room environment and CD procedures. We previously tested this Brief Exposure intervention in the Operating Room (BE-OR) among a small group of pregnant patients anticipating CD at a fetal care center and found a significant reduction in PMATS in the short (1-4 weeks) and the long term (6-8 weeks). These results are incredibly promising, but scalability to a large labor and delivery (L&D) unit with a more diverse patient population has not been established. To revise implementation plan and BE-OR for use in large L&D units with a broader population of patients, a fully powered multisite randomized control trial (RCT) is needed. Prior to initiating such a trial, Stage 1A/1B work needs to be done to modify the intervention and implementation through a process of iterative refinement to enhance acceptability, appropriateness, and feasibility of implementation in L&D units across the country, as well as its efficacy at engaging with the target mechanism (anxiety sensitivity). Using a logic model to guide the iterative refinement process through fast feedback loops and an atmosphere of co-creation, study investigators will gather critical input from stakeholders (individuals with lived experiences, community partners, front-line clinicians, hospital staff) via 12 workgroups, 12 user-testing design sessions, and repeated engagement with a steering council. Following this refinement process, a treatment development pilot RCT (N=80) at a large L&D unit will assess the efficacy of BE-OR by probing engagement with the target mechanism, as well as assess feasibility of implementation. Finally, the updated logic model and pilot trial results will inform the development of a protocol for a multisite RCT through engagement with expert consultants in a community engagement studio and further feedback from the steering council. This study will be the first in a long line of research broadening to additional risk groups, perinatal care settings, and intervention modalities to reduce PMATS.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY/ ABSTRACT Heart replacement therapies (HRT) (i.e., heart transplants and left ventricular assist devices [LVADs]) can improve quality of life and survival in patients dying of advanced HF. The patient selection pathway for receiving HRT is heterogeneous between centers with no universal consensus, increasing inequities and limiting access to potentially life-saving therapies. The medical evaluation for HRT is standardized, but the psychosocial assessment is different at each center, carries significant weight for candidacy, and is a frequent cause for patients’ ineligibility for HRT. During the psychosocial evaluation, inadequate duration of caregiving post-HRT is the most common perceived contraindication to candidacy. Caregiving requirements in the U.S. currently have clusters of centers requiring up to 1 month, 1-3 months, or 3+ months. No evidence exists to identify an ideal duration of caregiving to optimize patient outcomes. Variability in caregiver requirements creates healthcare inequities, particularly among vulnerable populations where socioeconomics can affect the ability for caregivers to devote longer durations of time away from work and/or child rearing. However, these variations are also understandable since there are tradeoffs associated with a decrease in caregiving requirements, including increased clinical staff burden after HRT. Per consensus, longer caregiving durations create institutionalized structures that exacerbate inequitable access to life- saving therapies, and 3+ months of continuous caregiving is considered “excessive” because centers with lower requirements still report optimal clinical outcomes. Given a lack of true empirical basis for caregiving standards and the willingness of some centers to create flexible, inclusive, creative solutions for patients whose social resources are limited, our ultimate goal is to conduct an R01 type 1 hybrid effectiveness- implementation trial that will: (1) test non-inferiority of shorter duration caregiving (1-month) at centers that currently require 3+ months of caregiving and (2) create an evidence base to define associations between caregiving and outcomes. However, changing center-specific caregiver rules will require significant planning, resources, and adaptations prior to implementation. This grant proposal uses the PRISM implementation science framework to conduct critical pre-implementation work. We will assess best practices and organizational processes needed to decrease caregiving requirements using the positive deviance approach (Aim 1), and will use implementation mapping to identify and adapt interventions that will allow centers to shorten caregiving requirements while simultaneously limiting clinical staff burden and maintaining optimal clinical outcomes (Aim 2). The goals of this research are consistent with the NHLBI’s strategic goals: (1) to investigate factors that account for differences in health among populations and (2) to optimize clinical and implementation research to improve health and reduce disease and inequities in care. The pre- implementation work proposed is essential for center-level changes that will be adaptable, acceptable, feasible, and sustainable as preliminary data for an R01 application.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY/ABSTRACT The rise of type 2 diabetes (T2D) in U.S. youth is an alarming public health issue. Although obesity is a strong risk factor for pediatric T2D, at the same time, only a subset of youth with obesity develops T2D, suggesting disease pathogenesis is driven by factors beyond total adiposity. It is well-established that altered body fat trafficking into abdominal and hepatic fats is strongly correlated with metabolic dysfunction in youth, independent of obesity. Recently, pancreatic traits assessed by imaging, particularly pancreatic fat and volume, have also emerged as potential risk factors for dysglycemia in youth. However, research examining the metabolic effects of these traits in youth has been limited to smaller cross-sectional studies, primarily conducted in clinical populations (i.e., children with existing obesity, fatty liver, or dysglycemia). Also, few, if any, human studies have explored the influence of the early nutritional environment on these pancreatic traits. In the proposed K99/R00 award, we will address these knowledge gaps by leveraging data from the Healthy Start Study, a longitudinal, pre-birth, cohort study in Colorado that has comprehensively characterized ~1,400 mother-child pairs since pregnancy. The applicant, Dr. Catherine Cohen, PhD, RDN, will add quantitative magnetic resonance imaging (MRI) sequences to scans performed at the next wave of visits when offspring are in adolescence (10-15 years) to measure pancreatic fat and volume. This data will then be used to pursue the following specific aims: Aim 1. Assess pancreatic fat and volume in a general risk cohort of adolescents and examine interindividual variation in relation to child characteristics and adiposity measures (K99). Aim 2. Test prospective associations of early nutrition/growth with pancreatic fat and volume in adolescence (K99/R00). Aim 3. Examine associations of pancreatic fat and volume trajectories across adolescence with trajectories of glucose-insulin markers across adolescence, and the role of pancreatic traits in mediating associations of early nutrition and growth with glycemic markers (R00).Taken together, the findings of this proposal will improve knowledge of the role of pancreatic traits as risk markers for early onset dysglycemia during puberty and generate insights that will enhance our ability to design and target T2D prevention strategies more effectively. The proposed training goals for the K99 phase will complement this research and focus on gaining: (1) experience applying lifecourse epidemiology concepts, (2) expertise in assessing body composition, especially adiposity measures, and (3) knowledge of pediatric T2D pathophysiology and dynamic glucose-insulin assessments, and will be achieved through individual mentorship, practical experiences, and didactic training, and supplemented with opportunities to expand professional development skills. Collectively, the proposed research and training activities will strategically equip the applicant with the knowledge/skills needed to transition from mentored (K99) phase to independent (R00) phase.

NIH Research Projects · FY 2026 · 2024-05

ABSTRACT In the United States (US), women account for an estimated 19% of new HIV diagnoses. The best way to reduce HIV incidence in all people (including women) is to link them to HIV pre-exposure prophylaxis (PrEP), which can reduce HIV transmission by up to 86%, with optimal adherence. The FDA approved the first long-acting form of PrEP, injectable cabotegravir (CAB-LA), in late 2021, which has the potential to decrease HIV transmission, increase PrEP adherence, and promote choice about HIV prevention for women. The addition of CAB-LA to available PrEP options necessitates women and PrEP service providers select the best method (oral vs injectable) for each person. However, women have unique concerns about PrEP (e.g., concerns about pregnancy/lactation) and report that patient/provider discussions on this medication are suboptimal. Thus, to inform this shared decision process, the proposed study builds on formative work by developing and pilot-testing “WePrEP,” a PrEP-focused bilingual digital shared decision-making tool (SDMT), tailored to women in the US and PrEP service providers. WePrEP will facilitate autonomy in HIV prevention choice by supporting communication between women and PrEP service providers as they identify their ideal PrEP product and discuss associated adherence strategies. This will occur by cuing conversations on women’s unique PrEP needs/concerns and presenting pertinent information that is relevant and appropriate. To develop and test WePrEP, we will partner with the Mile High Behavioral Healthcare, a Denver-based mental health and HIV prevention organization. We will use McNulty et al.’s adapted Shared Decision-Making Model to guide the iterative participatory design process we will use with a group of women and PrEP service providers to develop WePrEP; we will begin this process using prototypes created from preliminary data (Aim 1a). We will then rigorously assess the usability of WePrEP via simulated patient/provider discussions (Aim 1b). Next, we will pilot test WePrEP in a randomized controlled trial (RCT; N=69) with 2:1 randomization. In the RCT, PrEP service providers (N=4, of which n=1 is bilingual) will use WePrEP with intervention women to select CAB-LA or oral PrEP and discuss adherence. Other providers (N=2, of which n=1 is bilingual) will give control participants a standard of care explanation of PrEP (CDC recommendations) to help them make their PrEP choice. Women will be referred to Sheridan Health Services to start PrEP (if they wish; we acknowledge and respect that some participants will decide that PrEP is not right for them). We will assess primary (feasibility; acceptability) and secondary (potential mechanisms of action of WePrEP; preliminary impact) outcome measures using validated scales and rigorous qualitative methods (Aims 2, 3). By creating a bilingual digital SDMT to facilitate patient autonomy by enhancing communication between women and PrEP service providers as participants choose their ideal PrEP modality, our project is likely to make a widespread and lasting impact on uptake and adherence to PrEP, and HIV prevention independence. Our product is a rigorously designed, pilot-tested, bilingual, digital SDMT, suitable for use with all women in the US, that will test for efficacy in a larger, longer R01-funded RCT.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY Hypertension (HTN) is a primary risk factor for cardiovascular disease (CVD). The Systolic Blood Pressure Intervention Trial (SPRINT) was performed in hypertensive individuals with high CVD risk. The study showed that blood pressure (BP) target of <120mm Hg compared with <140mm Hg significantly reduced CVD events and all-cause deaths [1]. However, almost 15% of all SPRINT participants experienced serious adverse events (SAEs) including hypotension, syncope, bradycardia, acute kidney injury (AKI), and electrolyte abnormalities [1]. Our data point towards the contribution of the endocannabinoid system (ECS) in this process. EC system is an ancient, preserved, but only recently discovered biological system shown to play a pivotal role in regulation of BP [2]. The ECS is composed of endocannabinoids, including anandamide (AEA), the enzymes that produce and degrade endocannabinoids and their receptors. Interestingly, similarly to subjects that reported SAEs within the SPRINT trial, acute cannabis users present with bradycardia, hypotension, syncope and electrolyte abnormalities [3, 4]. Supporting studies in hypertensive animals have indicated the essential role of AEA-mediated signaling in the development of bradycardia and hypotension [5-10]. Interestingly, renin- angiotensin system, which is the main target of antihypertensive therapy, seems to be reciprocally regulated with the ECS e.g. increase in AEA reduces vasoconstrictive and hypertensive effects of angiotensin II [11, 12]. In our previous data, we showed reduced plasma levels of AEA in hypertensive non-CKD subjects and CKD as well as autosomal dominant polycystic kidney disease (ADPKD) patients as compared to healthy subjects [13]. Furthermore, intensive BP control in ADPKD patients (SBP <110mm Hg) as performed within the HALT-PKD trial, led to an increase of plasma levels of AEA and its congeners [13]. Based on these data, we hypothesize that intensive (<120mm Hg) as compared to standard (<140mm Hg) BP control in SPRINT participants will be accompanied by higher levels of AEA and its congeners. We also hypothesize that subjects with a greater rate of AEA increase will present with a higher incidence of SEAs including hypotension, syncope and bradycardia. The knowledge gained from this first-of-its-kind study will allow to better understand the role of the EC system in regulation of the blood pressure. With the increased use of cannabis and its active ingredients among general population, the understanding of the interplay between the EC system and the cardiovascular function will be of utmost importance for current patient management and future drug development studies.

NIH Research Projects · FY 2025 · 2024-05

Project Summary Hearing loss is one of the most common problems associated with normal aging. Sensorineural hearing loss, caused by damage or loss of inner and outer hair cells and characterized by raised hearing thresholds in older subjects, is well known and can be diagnosed through audiograms or otoacoustic emissions (OAEs). However, there is a subtype of hearing loss known as central or “hidden” hearing loss that similarly affects aged individuals. Hidden hearing loss is so termed because those afflicted present with hearing deficiencies, but have normal audiograms; their hearing problems are therefore “hidden” from normal diagnostic measures. Although subjects with hidden hearing loss have normal hearing sensitivity, they struggle to detect and understand sounds presented above a background noise, and struggle with localizing sounds in space. The underlying cause of hidden hearing loss is not currently well known, but because of the common symptoms of decreased sound localization abilities with spared hearing thresholds, it is thought in part to arise from dysfunction in the auditory brainstem. We hypothesize that hidden hearing loss in aging subjects is a result of dysfunction in the auditory brainstem, specifically a mistiming of the inhibition arising from the medial nucleus of the trapezoid body (MNTB). To test this hypothesis, we will perform auditory measurements in Mongolian Gerbils, an animal model commonly used in binaural hearing research because of its hearing range similar to humans and their ability to utilize interaural time differences (ITDs) and interaural level differences (ILDs) as humans do. We will perform auditory measurements such as OAEs, audiograms, and monoaural and binaural auditory brainstem responses (ABRs) and envelope following response (EFRs) to assess both the peripheral hearing and binaural processing abilities of young and aged gerbils. We will also perform behavior tasks to determine the spatial hearing abilities of these cohorts and correlate changes in behavior with potential biomarkers of spatial hearing from our ABR results. Additionally, we will investigate if age-related dysfunction arises from age-related changes directly to the auditory brainstem, a reduction of input from peripheral areas of the auditory pathway, or both. To assess this, we will perform in vivo recordings from auditory brainstem nuclei to directly measure the firing properties of neurons from the MNTB. We will also examine levels of cochlear synaptopathy, a reduction in the synapses in the cochlea to the auditory nerve, in young and aging animals as a metric of peripheral hearing ability. We expect to see a decrease of temporal fidelity of inhibitory neurons from the MNTB in aging animals, which would correlate with an increase of cochlear synaptopathy, a reduction of ABR waveforms confirming synaptopathy, and a decrease of spatial hearing performance in these animals. Taken together, our electrophysiological, immunohistochemical and behavioral measurements will provide a comprehensive understanding of the mechanism that drive age-related hidden hearing loss.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY Cannabis use during pregnancy is increasing with its legalization in many states and the belief by many that it is a safe and effective treatment for pregnancy-related nausea. This proposal investigates to what extent prenatal maternal supplementation with phosphatidylcholine (PChol) mitigates the effects of prenatal cannabis exposure on the fetus and subsequent child behavior. Higher maternal levels of choline have been shown to improve brain development. Choline in the amniotic fluid activates fetal alpha7-nicotinic receptors, before cerebral cholinergic innervation fully develops, late in gestation. Alpha7-nicotinic receptors and cannabinoid CB1 receptors are expressed on the same cerebral interneurons. Choline activation promotes interneuron development, whereas THC, by interfering with endogenous signaling, retards their development. Consistent with this translational model, a preliminary clinical observational investigation found that mothers with higher gestational choline levels have newborns with more normal fetal development of cerebral inhibition, even if mothers continued moderate cannabis use during pregnancy despite clinical advice. The child’s subsequent ability to pay attention is increased. We propose a randomized, double-blind, placebo-controlled trial of phosphatidylcholine supplementation in 120 pregnant women who use cannabis during pregnancy. Pregnant women will be randomly assigned to receive (1) 7200mg PChol, equivalent to 1028mg choline (Treatment Group, n=60) or (2) corn oil placebo (Placebo Group, n=60). The first aim of this project tests the hypothesis that maternal choline supplementation will interact with maternal cannabis use on the P50 auditory evoked response inhibition, an electrophysiological marker of the development of inhibitory neurotransmission, one month after birth. Aim 2 tests the hypothesis that maternal choline supplementation will interact with maternal cannabis use to mitigate its adverse effects on offspring’s development of attention, temperament, and cognitive behaviors. Prenatal cannabis use, assessed from mothers’ self-report and urine and serum metabolite levels, returned pill count, estimates of choline intake from the mother’s recall of her diet, and plasma levels of choline and its metabolites will be obtained at 14, 16, 22, 28, 34 and 40 weeks of gestation. Cannabis use in the postnatal period and breast milk cannabinoid and choline levels will also be measured. Neonate and infant hair will be analyzed for cannabinoids to detect 3rd trimester and postnatal exposure. The two aims together will document the possible effectiveness of prenatal choline supplementation to protect the fetus’s brain development from exposure to cannabis during gestation. The outcomes of this proposal could have an innovative public health effect to protect the brain development of fetuses whose mothers continue to use cannabis despite medical advice to abstain. These effects may result in decreased risk of mental problems in the children.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY / ABSTRACT The purpose of this K23 Mentored Patient-Oriented Research Career Development Award is to provide the applicant with advanced training to launch an independent research program focused on improving the effectiveness of parenting interventions for early childhood populations. Parenting interventions are the first line of treatment for early childhood mental health concerns due to their benefits on parenting stress and practices as well as most child emotional and behavioral problems. However, parenting interventions have substantial engagement issues, and many children show limited to no benefit. The objectives of this K23 are to (a) identify predictors of suboptimal response to a parenting intervention with well-established efficacy (i.e., behavioral parent training [BPT]) and (b) to identify ways to modify parenting interventions to benefit those parents, guided by their input. The central hypothesis is that greater parental executive functioning (EF) difficulties will be linked to more adverse social determinants of health (SDoHs), and both will diminish parent and child responses to a brief parenting intervention delivered through telehealth. Parents of children 3 to 7 years with externalizing concerns will be recruited from two sites – a children’s hospital and a group of Head Start programs – to receive BPT from trained clinicians. Multiple methods will be used at baseline to measure candidate predictors (i.e., parental EF, adverse SDoHs) in ecologically valid ways. A mixed methods approach will integrate qualitative and quantitative results and guide identification of potential adaptations to parenting interventions needed to achieve their intended benefits. The research project aims to (1) develop and test an ecologically valid measure of parental EF during parent-child interactions, (2) characterize the roles that parental EF and SDoH have on treatment response for parent (i.e., parenting stress, harsh parenting) and child outcomes (i.e., externalizing concerns), and (3) identify potential adaptations to improve intervention effects among non- responders. Under the mentorship of a high-quality team including Drs. Laura Anthony, Tina Studts, and Susan Mikulich, the applicant will acquire advanced training in the following areas: 1) acquire skills in developing and adapting real-world measures to use in clinical trials, 2) advance knowledge of how SDoHs influence parenting interventions, 3) gain competencies in mixed methods research, and 4) learn clinical trial methodologies. This project and training will enable the PI to become an independent scientist focused on discovering ways to improve the effectiveness of parenting interventions. The proposed training plan and research aims will provide the PI a foundation to conduct a large-scale trial (R01) testing if tailoring delivery methods (e.g., individual or group format; in-person or telehealth setting), dose (e.g., lengthen sessions), or content (e.g., target parental EF) improves the effectiveness of parenting interventions for non-responders. Findings will have significant implications for interventions that involve parents while treating youth mental health concerns.

NIH Research Projects · FY 2026 · 2024-05

Project Summary/Abstract Patients with hospital-onset sepsis (HOS) frequently receive delayed treatment and have nearly 3-fold greater risk of acute respiratory failure (ARF) and death than patients with community-onset sepsis, yet remain an understudied population. The current absence of data characterizing heterogeneity in clinical presentation, ARF risk, and treatment benefit, presents a critical barrier to designing interventions that improve treatment timing and prevent ARF for the most vulnerable patients. The candidate’s long-term career goal is to build an independently funded research program advancing precision approaches to HOS and ARF treatment. As an essential next step, the overall objective of this project is to create new knowledge about HOS heterogeneity and progression to ARF to inform the future development of treatment strategies tailored to patients’ individual organ failure risks and anticipated benefits from specific interventions. In preliminary studies, the candidate has built a retrospective cohort of HOS patients admitted to five organizationally diverse hospitals of an academic health system, and preliminary analyses have demonstrated an association between timing of antimicrobial administration for HOS and progression to ARF. This project will expand this cohort in order to complete the following aims: 1) identify latent phenotypes of HOS illness presentation using consensus k-means clustering accounting for informative missingness and including baseline patient factors, measures of organ dysfunction and severity of illness, and measures of degree and rapidity of clinical deterioration, 2) develop a novel model to predict ARF among patients with HOS using super learner machine learning predictive modeling including patient factors and physiologic data at sepsis onset; and 3) assess the association of antimicrobial treatment delays with progression to ARF among patients with HOS using inverse probability of treatment weighting to account for confounding by indication, and estimate heterogeneity of this effect by underlying ARF risk. To successfully complete these aims and acquire the necessary skills to transition to research independence, the candidate has developed a training plan that includes formal didactic training, research seminars, works in progress, and scientific meeting presentations focused on methods in machine learning, predictive modeling, and causal inference. Her career development will be supported by close guidance from her mentorship team who have deep content and methodologic expertise in relevant fields for this work.

NIH Research Projects · FY 2026 · 2024-04

Abstract: Heart failure (HF) is an expanding health problem, affecting >6.2M Americans, causing >80K deaths and >800K hospitalizations annually, and costing >$30B each year. HF is genetically complex as there are single- gene Mendelian (one mutation → causes disease) and disease susceptibility (several variants → influence risk of disease) models. The broad, long-term objective of our project is to use an integrated analysis approach to precisely quantify the contributions of rare Mendelian and more common disease susceptibility variants in dilated cardiomyopathy (DCM). Our project is responding to NOT-HL-23-067 (reissue of NOT-HL-21-017), Integrated Omics Analysis of NHLBI TOPMed Data, and targets the following key knowledge gaps: 1) how do rare Mendelian-disease and common complex-risk gene HF variants overlap and interact in the pathogenesis of HF, 2) which complex risk-variants are most likely to be functional (i.e. manifest specific biological mechanisms), and 3) what additional HF genes remain to be discovered (~50% of genetic HF lacks a pathogenic variant(s)). Our approach will leverage a unique whole-genome and whole-transcriptome human heart NHLBI- sequenced TOPMed dataset (~750 paired DNA/RNA sequenced human samples) using an integrated analysis approach to dissect the intersection of Mendelian and complex genes and variants. Building on these resources and approaches we will 1) validate which HR GWAS variants actually affect gene expression in the human heart, 2) discover novel human heart expression Quantitative Trait Loci (eQTL) from the cardiac transcriptome in health and disease, and 3) discover novel HR genes and pathways. Our hypotheses are 1) a subset of published GWAS HR variants exert direct eQTL mechanistic effects on gene expression in human cardiac tissue, 2) key genes and pathways of the HR transcriptome are mediated by novel genetic variants that have heretofore not been identified by GWAS approaches, and 3) integration of published Mendelian and GWAS HR variants in combination with cardiac tissue eQTL variants will reveal important, novel HR genes. Three Specific Aims to test these hypotheses are: 1) Map and validate published GWAS findings onto the human cardiac transcriptome, 2) Discover novel eQTL DNA variants from HR transcriptomes, and 3) Discover novel HR genes and perform functional validation studies in human heart tissue and hiPSC- cardiomyocytes. The impact of these studies will be a comprehensive genetic and RNA expression atlas of GWAS and eQTL biology in the human non-failing and failing heart models. This work will reveal and validate novel HR genes and pathways for the development of HR biomarkers and new treatment approaches.