University Of Colorado Denver

NIH Research Projects · FY 2025 · 2022-05

Abstract Allergic diseases including food allergies and allergic asthma represent a substantial health and economic burden, with the US Centers for Disease Control (CDC) estimating more than 50 million people within the US suffering from some form of allergic disease. The natural history of allergic diseases suggests immunological crosstalk between mucosal organs contributes to the co-occurrences of allergic diseases (e.g. food allergy and asthma) within the same individual, although the cellular and molecular mechanisms that underly this process have not been defined and represent a critical knowledge gap. Using in vivo mouse models we have shown that intragastric administration of allergen (ovalbumin, OVA) to OVA-sensitized mice not only elicits local eosinophilic inflammation within the allergen-exposed intestine, but also increases the frequency and alters the activation phenotype of tissue eosinophils within the allergen non-challenged, remote lung. Presence of inflammatory eosinophils within the remote lung are associated with mucous metaplasia and airway priming; the latter evidenced by generation of an exacerbated allergic airway inflammatory response upon subsequent inhalation of sub-optimal doses of an unrelated antigen (house dust mite). In contrast, intragastric OVA failed to enhance mucous metaplasia or airway priming in the remote lungs of eosinophil deficient mice. Collectively these prior data suggest intragastric challenge affects allergic susceptibility of the airways through dysregulation of lung tissue eosinophils. This proposal builds on these findings to investigate lung- and eosinophil-intrinsic mechanisms that underly intragastric allergen-driven dysregulation of remote lung eosinophils that lead to remote airway priming. We have found intragastric allergen challenge transiently alters the lung transcriptome, including induction of gene signatures implicated in eosinophil recruitment. Correlative analyses of serum proteins and blood eosinophil-expressed receptors further suggest systemic mediators and eosinophil-intrinsic factors contribute specifically to lung homing, including an eosinophil-derived subset of extracellular vesicles (EVs). Specific Aims test the central hypothesis: Intragastric allergen challenge activates 1) an IL-13:CCL11 axis in the remote lung via gut-derived type 2 innate lymphoid cells (ILC2s), and 2) IL-5/IL-33:ST2-dependent effects on circulating eosinophils that synergistically underly the dysregulation of tissue eosinophils within the remote lung. Our approach utilizes unique genetically modified mouse strains, including cytokine reporter mice and cell-targeted gene disruption, competitive adoptive transfer studies and innovative approaches to characterize eosinophil-derived extracellular vesicles. Translational approaches utilize human blood eosinophils. Completion of this proposal may offer important insights into immunological mechanisms that drive mucosal remote organ priming within the context of allergic inflammation, and as such is highly relevant to the development of better diagnostic and interventional approaches for patients with allergic diseases.

NIH Research Projects · FY 2026 · 2022-04

Research Summary The prevalence of alcohol use disorders (AUDs) in the U.S. is approximately 12.7% and has shown a marked increase in recent years (Grant et al., 2017). Individuals with AUD often use cannabis, which may impact attempts to reduce or quit drinking. In the last six years, the United States has witnessed enormous changes concerning the public acceptance and availability of cannabis. Considering that cannabis has historically been the drug most often used by individuals with an AUD and considering the skyrocketing increase in availability of cannabis products, it is imperative for health care providers to understand how best to approach cannabis use with individuals who have an AUD and who want treatment. The central premise of the proposed research is that products with low THC and high CBD may be less harmful to AUD individuals who want to quit drinking, as compared to products with high THC only. It is hypothesized that individuals who switch from a high THC product to a low THC and higher CBD product will demonstrate superior outcomes compared to those individuals who do not switch. If the hypotheses are supported, the dissemination of the research would have an immediate effect on public health impact by educating patients and treatment providers about how to address the use of cannabis among individuals who want to quit or reduce alcohol consumption. To that end, individuals who currently use a high THC product and want to reduce or quit drinking will be randomly assigned to either the control condition (continue to use your current cannabis product as you wish) or the intervention condition (instruction to switch to low THC/high CBD product). Participants in the intervention condition choose the product and use as little or as much as they want during the study. Participants will be tested in the Mobile Pharmacology Laboratory at 6 and 12 week timepoints to determine the impact of these products on cue-elicited anxiety, cue-elicited alcohol craving, drinking outcomes, and biological markers of systemic inflammation.

NIH Research Projects · FY 2026 · 2022-04

ABSTRACT: The overall goal of this proposal is to understand how the gain-of-function MUC5B promoter variant affects transcriptional regulation and gene expression that are common to clinically distinct types of usual interstitial pneumonia (UIP). UIP was initially described as a morphologic entity, however, recently has been defined using specific pathologic and radiographic criteria. While UIP is characteristic of idiopathic pulmonary fibrosis (IPF), these pathologic and radiographic patterns of chronic fibrosing interstitial pneumonia are also typical of chronic hypersensitivity pneumonitis (CHP), rheumatoid arthritis-associated interstitial lung disease (RA-ILD), asbestosis, and several drug-induced lung diseases. The gain-of-function promoter variant in MUC5B (rs35705950) is the dominant risk factor for IPF, is present in >50% of affected patients, and has also been reported to be the dominant genetic risk variant for the development of CHP and RA-ILD. However, while IPF is by definition idiopathic, CHP develops following repeated exposure to organic antigens, and RA-ILD is a complication of rheumatoid arthritis. We proposed by understanding the relationship between the MUC5B promoter variant, transcriptional regulation, and gene expression in IPF, CHP, and RA-ILD, we will be able to identify the common molecular elements that are critical to the development of UIP. The two critical questions that we plan to address are: 1) what are the common molecular features of UIP, irrespective of clinical context, and 2) does MUC5B promoter variant have a unique molecular signature common to UIP? The hypothesis we plan to test is that the gain-of-function MUC5B promoter variant drives cell-specific chromatin accessibility and gene expression that define UIP. In Aim 1, we will use single nucleus RNA sequencing (snRNA-seq) to identify the cell-specific transcriptional profiles for IPF, CHP, and RA-ILD, and determine the relationship of the common cell-specific transcriptional profiles of UIP to the MUC5B promoter variant. In Aim 2, we will use a combinatorial indexing single nucleus assay for transposase-accessible chromatin (snATAC-seq) to identify the cell-specific chromatin accessibility profiles for IPF, CHP, and RA-ILD, and determine the relationship of the common cell-specific chromatin accessibility profiles of UIP to the MUC5B promoter variant. In Aim 3, we will perform integrative analyses of the MUC5B promoter genotype, snRNA-seq, and snATAC-seq data using single-cell expression QTL, multi-omic and network inference methods to identify UIP-specific gene regulatory networks with key drivers of UIP in specific cell types. In Aim 4, we will validate the transcriptional features that are common to MUC5B-associated UIP by determining the relationship of these key genes to the pathogenic heterogeneity of UIP and relevant in vitro biology. In aggregate, we will characterize regulatory effects of MUC5B on cell-specific transcriptional profiles and networks in UIP, launching investigation of novel pathogenic mechanisms and drug targets for these incurable diseases.

NIH Research Projects · FY 2026 · 2022-04

The ultimate goal of this proposal is to define composite biomarkers that can be used to improve outcomes in future type 1 diabetes (T1D) clinical trials. T1D is the major cause of diabetes in youth. It is characterized by life-long insulin insufficiency due to autoimmune mediated ß cell destruction. Despite considerable efforts over the past 30+ years, effective therapies are still lacking and there is an urgent need for a cure. Natural history studies indicate that the rate of T1D progression varies greatly between individuals, both before, and after onset. Indeed, the current paucity of validated mechanistic biomarkers that can accurately predict “slow” or “fast” progression is a major impediment to finding a cure. At least 40-60% of patients experience a period of partial remission (PRM) in the first 6 mo after they begin taking insulin. This “honeymoon period” is highly variable, ranging from a few weeks to several years. Like T1D, the factors that govern the onset and duration of PRM are not fully understood. Initially it was believed that PRM is solely a metabolic phenomenon, but there is increasing evidence that the immune system also plays an active part. This leads to the primary hypothesis that underpins our proposal: identification of immunological, metabolic, and demographic features that associate with PRM duration will enable the development of improved clinically actionable composite biomarkers for T1D. Our study has a single specific aim, namely, to define and validate one or more classifiers that can accurately predict fast or slow progression of T1D in the first 2y post-onset from baseline data. This will be achieved through an in depth multimodal analysis of peripheral blood drawn from a cohort of 100 subjects with a recent diagnosis of T1D. A single draw will be made at 3-6 months post diagnosis, and a range of assays performed with DNA, RNA, protein and functional readouts, and ranging in complexity from single analytes to single cell transcriptomes. PRM duration will be determined from clinical data collected over the following 1.5-2y. Subjects will be randomized to training and validation cohorts matched for age, gender, and content of “fast” and “slow” progressors. Features from the analytical data will be used to generate models that predict PRM duration using DIFAcTO, a machine learning algorithm that combines univariate filtering, hierarchical clustering, and LASSO regression, to select non-redundant features that result in an optimal model. Performance of the final models will be evaluated by applying them to the independent validation cohort. The features retained in the resulting models will be prime candidates as composite biomarkers to improve subject stratification at recruitment, and aid identification of responders and non-responders, in future clinical trials. Thus, if successful, our study should have significant impact on the field.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY Acetaminophen (APAP)is one of the most commonly used analgesics in the world, andoverwhelmingly perceived to be safe. This perception has contributed to ubiquitous exposures during gestation and among newborns in the neonatal intensive care unit. These exposures occur during critical windows of human development where pre-clinical and clinical data demonstrating safety are lacking. Alarmingly, clinical data support the hypothesis that the developing lung may be adversely affected by perinatal APAP exposures. The toxicity of APAP is dependent on its conversion by the xenobiotic-metabolizing enzyme CYP2E1 the mitochondrial toxin N-acetyl-para-benzo-quinone imine (NAPQI). Our preliminary data demonstrate that pulmonary CYP2E1 expression peaks during the saccular stage of development and is limited to the myofibroblast. Additionally, we show that postnatal APAP exposures induce Cyp2e1 expression in the late saccular/early alveolar stage lung. Consistent with CYP2E1 expression, we show that the developing lung is susceptible to APAP-induced injury. These preliminary data have led us to develop the following hypothesis: The saccular/early alveolar stage lung is susceptible to APAP-induced injury due to developmentally- regulated pulmonary CYP2E1 expression. We propose three specific aims to test this hypothesis. In Aim 1, we will test the hypothesis that in utero APAP exposures during the saccular stage of lung development injure pulmonary myofibroblasts and disrupt alveolarization. In Aim 2, we will test the hypothesis that postnatal APAP exposures during the late saccular/early alveolar stage induce lung CYP2E1 expression causing oxidative stress and increase sensitivity to injury. In Aim 3, we will test the hypothesis that inhibiting TLR9/NFκB innate immune signaling will attenuate APAP-induced newborn lung injury. Our collaborative team bridging developmental pulmonary biology, physiology, immunology and mitochondrial/oxidative stress biology is well positioned to fill critical gaps in our understanding of the developmentally-regulated, cell-type specific CYP2E1 expression and APAP-induced lung injury. These studies will help determine the safety profile of APAP to inform both maternal use and newborn exposures while identifying therapeutic targets to limit adverse effects. into

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY Pruning neuronal connections and eliminating superfluous neurons is required to generate optimized neuronal circuits in the mature brain. Although it is well established that neurons and glia coordinate the refinement of neural circuits, the molecular mechanisms underlying this process remain poorly defined. This K99/R00 proposal will help me advance my career as I investigate the cellular mechanisms by which glia help refine neuronal circuits during development. To generate a deep molecular understanding of neuronal remodeling, I will use the Drosophila larval nervous system, which goes through extensive neuronal remodeling during metamorphosis. In a preliminary screen for new model cells to study neuronal remodeling in vivo, I discovered several new markers for cells that exhibit novel types of remodeling events. In addition, I conducted two large-scale screens to identify new glial pathways that assist in pruning and the elimination of neuronal debris. First, using a single neuronal lineage, I performed an in vivo RNAi screen for glial genes required for glial pruning of neurons. Second, I transcriptionally profiled glia during pruning, identifying upregulated genes in glia, and also screened them for regulators of neuronal remodeling. These genes will serve as a molecular entry point for me to define how glia refine neural networks during development. During the mentored award phase, I will build a system that will allow me to monitor the dynamic cell biological changes that occur during glial phagocytosis and will use this new system to test novel molecules for their role remodeling. In Aim1, I will study the specific process of glial phagocytosis using genetically encoded cellular markers and explant live imaging. These assays will serve as the foundation for understanding how the molecules I identified play a role in glial phagocytosis. In Aim 2, I will use this new system to understand how Tweek, a highly conserved molecule, functions during glial phagocytosis of pruned neurons. Surprisingly, Tweek has no known protein domains or molecular function. I will use CRISPR/Cas9 genomic engineering to create human disease-associated mutations in the fly. This will potentially allow me to understand how mutations in Tweek's human homolog KIAA1109 cause a rare autosomal neurological disease. Finally, in Aim 3, which will be mostly carried out in my own lab, I will use the tools I build in this proposal to examine how a collection of newly identified phagocytic receptors drive neuronal remodeling of synapses, neurites, and in multiple types of lineages. I outlined a series of research and career development milestones that will be met during this award and allow me to grow as a scientist. To strengthen areas needed for a successful research career, the aims are combined with neuroscience coursework and training in areas such as scientific writing, mentoring, and project design. I have a committee made up of an excellent group of scientists who are dedicated to my success and eager to assist me in my professional development. My long- term career goal as an independent investigator is to understand the molecular mechanisms of neuronal remodeling and how abnormalities in this process are associated with neurodevelopmental disorders.

NIH Research Projects · FY 2026 · 2022-04

Project Abstract End-stage heart failure is a leading cause of death worldwide and costs the United States over $30 billion annually. Heart transplant remains the only curative therapy but its success is threatened when the donor heart does not work properly. Current immunosuppressive treatments focus on targeting immune cells in the recipient's heart. These therapies confer a significant risk of infections and malignancy and are only modestly effective. Targeting the donor-recipient interaction may provide a novel therapeutic pathway. Several distinct immune cell types reside within the donor heart with macrophages comprising the majority of myeloid cells. The mouse and human heart contain at least two populations of macrophages that can be distinguished based on the expression of C-C chemokine receptor 2 (CCR2). CCR2+ macrophages participate in inflammatory responses whereas CCR2- macrophages promote tissue repair. The PI has established that donor CCR2- and donor CCR2+ macrophages have opposing roles after heart transplant. Depletion of donor CCR2- macrophages accelerates rejection while depletion of donor CCR2+ macrophages prolong allograft survival. In this proposal, the PI will decipher the mechanisms by which donor CCR2- macrophages prevent rejection through their modulation of other immune cell populations and signaling pathways. The scientific goals of this research award are to identify novel immune populations that can be exploited to understand the mechanisms of rejection and can be therapeutically targeted to improve outcomes. By the end of the award period, the PI will understand how donor CCR2- macrophages effect donor CCR2+ macrophage activation and whether this activation is necessary for allograft rejection (SA1). The PI will investigate how CCR2- macrophages become activated and whether this activation is required for allograft protection (SA2). In completing these aims, the PI will gain technical expertise in flow cytometry, intravital imaging, bulk and single cell RNA sequencing, antigen presentation, phagocytosis, and alloreactivity assays. The career development goal of this proposal is to develop the PI into an independent physician-scientist in the field of cardiovascular research. The PI has previously obtained a PhD training in biology and has obtained additional training in basic and translational cardiovascular research during his post-doctoral fellowship. The PI has completed clinical training in Internal Medicine, Clinical Cardiology, and Advanced Heart Failure and Cardiac Transplantation Fellowship. The proposed 5-year career development plan will provide the PI with formal training in immunology and bioinformatics and ongoing laboratory training in the study of cardiac transplantation and macrophage biology. The PI will meet regularly with his mentors and advisory committee which is composed of senior scientists who are experts in immunology, transplantation, cellular imaging, bioinformatics, and career development. At the conclusion of this award period, the PI will have acquired the skills necessary to become an independent and successful physician-scientist.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY Cancer cell genomic plasticity can enable resistance to cancer therapy for both solid tumors and hematologic malignancy. Escape from cytotoxic or molecularly targeted therapy through an inherent capacity to reprogram differentiation state or lineage has now been described following adoptive cell therapy or immune checkpoint blockade in adult epithelial tumors. Transfer of T cells genetically modified to express chimeric antigen receptors (CAR T cells) targeting the B cell surface antigen CD19 induces remission in 70-90% of patients with relapsed/refractory B cell acute lymphocytic leukemia (B-ALL) resulting in FDA-approval for this indication. However, a large fraction of those patients relapse within one year of treatment. This occurs with two main patterns, 1) early antigen-positive (CD19pos) relapse, attributed to poor CAR T expansion or lack of persistence, and 2) later antigen-negative relapse. Evasion of CD19-targeted immunotherapy can result from loss of all B lineage phenotypic markers with acquisition of stable, alternative phenotypes in MLL-rearranged (MLL-r), BCR-ABL driven, TCF3-ZNF384 and other subtypes of ALL. Remarkably, emergence of phenotypic switch can occur years after CD19-targeted immunotherapy. Understanding the mechanisms of immunotherapeutic resistance and identifying strategies to overcome these will be critical in improving remission depth and durability of response. Our proposal will address two major deficits in cancer models to identify factors contributing to relapse from immunotherapy: the lack of immune-intact model systems that recapitulate the lineage switching phenomenon observed using CD19-targeted immunotherapy and the lack of faithful mouse models recapitulating infant/childhood MLL-r B-ALL. This collaborative proposal brings together the extensive expertise of the Ernst group in the biology of MLL-r leukemia and hematopoiesis with the CAR T cell expertise of the Fry/Kohler groups to develop innovative new models systems to study evasion from CAR T cell therapy through lineage reprogramming. Our preliminary CAR T cell data employs immune-intact mouse models to illustrate that CD19neg relapse includes cells that exhibit gain of myeloid antigens and a myeloid transcriptional profile. On the pediatric B-ALL front, we develop a retroviral system to produce B-ALL that captures the inherent plasticity of MLL-r leukemias and switches to AML in vivo. Our proposal assesses the ability for both leukemia-intrinsic as well as extrinsic host- environmental components to influence escape from CAR T killing through lineage reprogramming. The findings of our studies, including the discovery of novel strategies to block lineage reprogramming have the potential to inform the development of similar approaches in other forms of cancer treated with cellular therapy and, potentially, immune checkpoint inhibitors. In addition, these studies may lead to a better understanding of lineage plasticity and the extent to which epigenetic heterogeneity contributes to relapse, which can directly inform the design of curative therapies.

NIH Research Projects · FY 2025 · 2022-03

Artificial Intelligence (AI) - computer-based algorithms capable of learning from enormous data sets, including electronic health records and chart notes, in order to carry out tasks typically reserved for humans – is poised to dramatically affect medical research and practice, including end-of-life and palliative care (EOLPC). Recent AI-based algorithms seem capable of accurately predicting a patient’s prognosis or probability of death years in advance. These algorithms can do so in an automated fashion, without the input of clinicians, and they are starting to move from research into practice. For the millions of Americans who experience the physical, psychological, and social effects of severe and chronic illness, knowing a prognosis could promote earlier access to palliative care and to support medical decision-making that is consistent with patients’ and families’ goals and preferences. However, AI also raises concerns about loss of autonomy in patient or clinician decision-making, depersonalized or unempathetic care, distrust of “black box” machines, and an overemphasis on survival statistics in decision-making. Studies consistently show that patients and caregivers may be unaware of their prognosis, that physicians are often inaccurate in predictions, and that some patients may be less aware of their prognosis; however, the need for an accurate prognosis may vary by disease state, individual preference, or other sociocultural factors. Thus, how AI-based prognostication will affect our basic scientific understanding of the role of prognostic awareness in medical decision-making in support of high quality, goal concordant EOLPC is a critical knowledge gap. Before AI becomes more widely used in EOLPC, spreads to other uses (e.g., virtual nurse assistants and caregiver robots), or becomes necessary as proof of eligibility for services (e.g., hospice), there is an urgent need to understand its potential impact on patient- and family-centered care and to develop practical ethics guidance for its use. The goal of this project is to ensure AI is developed and implemented in ways that support high quality EOLPC. With a unique team of experts in palliative care, artificial intelligence, bioethics, and patient engagement, we will: (1) use semi-structured interviews to obtain rich insights into the experiences and beliefs of all EOLPC team members, patients, and family caregivers regarding AI-based prognostication at 4 purposefully chosen sites across the United States; (2) conduct a national survey of palliative care physicians regarding the anticipated benefits and challenges of using AI-based prognostication; and (3) convene a Delphi panel of experts to create practical recommendations for the use of AI in EOLPC. The project will be supported within the Palliative Care Research Cooperative Group (PCRC) (U2C NR014637), a robust interdisciplinary research community comprised of more than 500 members at more than 180 sites

NIH Research Projects · FY 2026 · 2022-03

Project Summary/Abstract Astrocytes are non-neuronal cells widely distributed in the brain and astrocyte-neuron communication play critical roles in modulation of behavior. Sensory stimuli like chemical signals of odors activate neurons to induce gene expression changes that facilitate sensory processing. While these are well understood in neurons, whether similar transcriptomic changes also occur in astrocytes are unknown. Here, using in vivo chemogenetic models of neuronal activation, we show astrocytes indeed undergo robust gene expression changes after neuronal activation. A screen through these changes identified a neuromodulator transporter Slc22a3 in olfactory bulb astrocytes. Since preliminary studies revealed odor-evoked neuronal activation also increased astrocytic Slc22a3 in the olfactory bulb, we first asked how Slc22a3 affect astrocyte function in the olfactory bulb? We show that overexpression of astrocyte-specific Slc22a3 led to increased sensitivity to odors implying that astrocytic Slc22a3 affects astrocyte-neuron communication. This leads to the hypothesis that transcriptional activation of astrocytic Slc22a3 is essential for mediating astrocyte-neuron communication during olfactory processing. To test this hypothesis, we propose to use Slc22a3 gain-of-function and loss-of- function mouse models to investigate how astrocytic Slc22a3 affect behaviors and cellular properties of astrocytes (Aim1). Since Slc22a3 transports neuromodulators like serotonin, we next asked how Slc22a3- mediated serotonin transport affect molecular properties of olfactory bulb astrocytes? Since recent studies have shown that serotonin can be directly incorporated into histones to activate transcription, we focused on this epigenetic modification of histone serotonoylation. We show that in olfactory bulb astrocytes, overexpression of Slc22a3 controls histone serotonoylation levels. Therefore, we propose to genetically manipulate astrocytic Slc22a3 expression to determine Slc22a3-mediated histone serotonoylation dynamics in the olfactory bulb (Aim2). Furthermore, preliminary data also revealed that histone serotonoylation levels are increased in astrocytes after odor-evoked neuronal activation. Therefore, using experimental approaches established in Aims1-2, I will directly investigate the function of astrocytic histone serotonoylation in the olfactory bulb (Aim3). Taken together, these results will reveal genetic and epigenetic mechanisms of how astrocytes contribute to olfactory processing. For my career development these studies will provide training in olfactory bulb biology under my mentor Dr. Deneen (expert in astrocyte biology) and co-mentor Dr. Arenkiel (expert in olfactory bulb circuits and behaviors) at Baylor College of Medicine. Since astrocytes are intimately connected with neurons and are dysregulated in all neurological disorders, the broader goal of this proposal is to uncover how astrocytes contribute to healthy processing of the chemical senses of smell. Towards this goal, the proposed research will apply new approaches of astrocyte biology to delineate genetic and epigenetic mechanisms involved in normal function of the smell system.

NIH Research Projects · FY 2025 · 2022-03

Abstract: Controlling the autoimmune inflammation in type 1 diabetes (T1D) has proven difficult. Currently there are clinical trials to attempt controlling T1D that are having only marginal impact. Desired primary outcomes include increasing of C-peptide, as a marker for beta cell restoration, decrease insulin requirements, decreases in HbA1c, as a measure of systemic inflammation and maintenance of peripheral blood lymphocyte counts. Current therapies have slowed C-peptide loss marginally but not halted or reversed loss. None of the other primary goals have yet been achieved in any of the current clinical trials. We created a novel approach to control pathogenesis in T1D using a small peptide to modulate CD40 mediated inflammation. We completed pre-clinical studies in mice that include toxicology/pharmacodynamic/pharmacokinetic studies on a peptide that prevents diabetes onset in NOD mice and reverses hyperglycemia in 60% of diabetic NOD mice. We have begun a veterinary clinical trial generating data that show, unlike current clinical trial treatments, this approach increases C-peptide over time, reduces, by up to 90%, insulin requirements, reduces glycated fructosamine, the veterinary equivalent of HbA1c, and does not cause immune suppression or lymphocyte loss. We were granted a “Safe-to-Proceed” notice from the FDA to begin Phase 1a/1b clinical trials in humans using this drug. This grant application is to perform mechanism of action studies on this drug. We hypothesize that the drug, KGYY15 (OPT101 for FDA) binds to beta cell CD40 to prevent beta cell damage. We further hypothesize that KGYY15 binds to peripheral blood T cells (TH40 cells specifically defined by us), B cells and macrophages/dendritic cells to tolerize thus preventing further damage. We also will explore developing this drug approach for islet transplants. Successful completion of this grant will provide important information about how this drug works specifically during T1D.

NIH Research Projects · FY 2026 · 2022-03

Abstract Despite major gains in smoking cessation treatment, over half of recently quit smokers will relapse within the first year. Two systematic reviews of relapse prevention studies came to different conclusions on effectiveness of behavioral interventions. Existing evidence in relapse prevention is limited by study designs, methodology, and conceptual approaches to behavioral interventions. Different approaches to relapse prevention studies, and to the interventions themselves are needed to advance the long-term understanding and outcomes of smoking relapse prevention. To date, relapse prevention interventions have focused on the newly abstinent smoker (“abstainer”), and not attempted to directly or indirectly influence the abstainer’s personal network (PN), e.g. by helping the abstainer influence others in their PN to quit. Personal networks exert powerful effects on initiating and maintaining smoking behavior, and can facilitate maintaining abstinence or trigger relapse. A “help others” intervention that seeks to increase the abstainer’s ability to influence smokers in their PN to quit – thereby creating a PN social environment more supportive of long-term abstinence - may have a beneficial effect on relapse. The Helpers SQ intervention encourages abstainers to reinforce their own abstinence through helping others quit, and to proactively influence their PN to be more conducive to long-term smoking abstinence. Framing relapse as a dynamic and complex process, the Helpers Stay Quit (Helpers SQ) intervention is a conceptually novel approach to relapse prevention that integrates different behavioral theories into a multifaceted intervention model presented as an on-line tobacco cessation brief intervention training. Helpers SQ teaches abstainers how to encourage other tobacco users to quit and avoid relapse through a non- confrontational “helping conversation” (HC) that encourages quitting and use of evidence-based cessation aids (e.g. quitlines, cessation medications) without confrontation and nagging. Our pilot feasibility study of Helpers SQ (N=104) with abstainers from Arizona’s state quitline compared 30-day abstinence at 7-months with a propensity score matched sample from quitline clients not exposed to Helpers SQ. Preliminary results: Helpers SQ participants reported higher 30-day abstinence than non-participants (82% vs. 36%, Difference = 46% [95% CI: 37%, 56%, p<0.0001]). Encouraged by these results, we hypothesize that quitline abstainers exposed to Helpers SQ will have higher 30-day and 7-day point prevalence abstinence than those receiving quitline follow-up usual care, and that the effect of Helpers SQ may be mediated by PN characteristics. To test this hypothesis, we propose a pragmatic randomized controlled trial (N=940) with embedded mixed-methods PN study to assess the effect of Helpers SQ training on proportion and duration of abstainers’ abstinence over time, and on abstainer’s PN interactions related to smoking and smoking cessation. Metrics derived from the PN study will be used for mediational analyses of overall, and gender-based effects of Helpers SQ on smoking relapse.

NIH Research Projects · FY 2026 · 2022-03

Project Summary/Abstract Inflammatory bowel disease (IBD) is a common disorder that causes inflammation predominantly in the digestive tract. Although treatment options have expanded in the past two decades, a significant portion of patients still suffer from poor treatment response and chronic clinical courses. Various responses to currently available therapy are in part due to the heterogeneity of pathogenesis, and novel mechanistic insight that could lead to new therapeutic options is critically needed. Intestinal intraepithelial lymphocytes (IELs) are a heterogeneous group of tissue-resident lymphocytes, and pathological activation of IELs is implicated in the exacerbation of IBD. However, the regulatory mechanisms of IELs during activation are not well understood. Aiolos, a transcription factor encoded by Ikzf3, is expressed exclusively in lymphocytes, and contributes to B cell activation and NK cell maturation. Aiolos mutations are implicated in various autoimmune diseases including IBD, but the roles of Aiolos in IELs or gut homeostasis have not been studied in the past. I discovered that Aiolos is highly expressed in IELs and that Aiolos-deficient IELs are more activated and express higher levels of activation markers (NK receptors) and cytokines (interferon-g and granzymes) compared to wild-type cells. These preliminary results indicate that Aiolos strongly suppresses IEL activation. Since Aiolos is a transcription factor modulating chromatin remodeling, I hypothesize that Aiolos binds to the enhancer regions of genes associated with IEL activation and controls their expression by altering epigenetic modifications. Additionally, since Aiolos potently regulates IEL activation, I hypothesize that Aiolos-deficient IELs modulate the host response to colitogenic infection and intestinal inflammation. In this proposed project, I will address these hypotheses with the following aims: (Aim 1) to identify Aiolos binding sites and define the contribution of Aiolos to epigenetic landscape in IELs, and (Aim 2) to investigate the impact of Aiolos-deficient IELs during mucosal inflammation. In Aim 1, I will utilize state-of- the-art epigenetic techniques to examine Aiolos binding sites in wild-type IELs and investigate how Aiolos deficiency alters epigenetic modifications in IELs. I plan to examine differences of epigenetic modifications between Aiolos-deficient IELs and wild-type IELs by comparing open chromatin regions and histone modifications. In Aim 2, I will challenge mice with Aiolos-deficient IELs and mice with wild-type IELs with Salmonella typhimurium and dextran sodium salt-induced colitis to delineate the impact of Aiolos-deficient IELs on host responses to intestinal infection and inflammation. Taken together, this project will contribute to profiling cis-regulatory circuits of IELs and could advance our understanding of the pathogenesis of IBD, which could eventually lead to the identification of new therapeutic targets.

NIH Research Projects · FY 2026 · 2022-03

SUMMARY CAR-T immunotherapy has great promise as a salvage regimen for patients who will no longer respond to conventional therapies. Five CAR-T cell therapies (four targeting CD19 on cancer cells) have been approved by FDA for treatment of relapsed and/or refractory (r/r) B-lineage malignancies, including ALL, Non-Hodgkin Lymphoma, CLL and multiple myeloma. However, about 50% of B cell leukemia and lymphoma patients treated with CD19 CAR-T therapy relapse within a year after CAR-T therapy. The success of CAR-T therapy has been associated with several factors, including: 1) the initial expansion of CAR-T cells after transfer into the patients in an IL-2-deprived environment, 2) maintenance of the CAR-T cell effector function 3) long-term survival of CAR-T cells. These T cell biology aspects are highly influence by their metabolic stage, and CAR-T cell outcome is known to be influenced by their metabolism. Metabolism is now considered as a major regulatory factor of the function of immune cells and influences the course of an immune response. We have identified MCJ (Methylation-Controlled J protein) as a protein localized in the inner membrane of mitochondria that acts as an endogenous negative regulator of Complex I and mitochondrial respiration (mitochondrial ATP production). We have shown that loss of MCJ in CD8 cells enhances cytokine secretion as well as cytotoxic activity. Memory MCJ KO CD8 cells have superior protective activity against influenza virus. MCJ deficient CD8 cells are also more efficient in killing tumor cells. Thus, we hypothesize that eliminating MCJ as a metabolic brake in CD8 cells will result in enhanced cytokine production, tumor killing activity and survival of CAR-T cells and that MCJ could be an attractive target to improve the success of CAR-T immunotherapy. To test this hypothesis and show its clinical relevance, we propose: 1) to evaluate the role of MCJ as a regulator of mitochondrial metabolism and effector function in human CD8 cells; 2) to evaluate the in vitro and vivo potency and efficacy of mouse MCJ-deficient CAR T cells, 3) to develop an MCJ-deficient human CD8 CAR-T with improved survival, expansion and cytotoxic activity.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY The development of cardiovascular disease (CVD) is a complex process, with obesity being a major risk factor. Weight loss among overweight/obese individuals is a key component of the primary prevention of CVD, but there is marked variability across individuals in response to lifestyle weight loss interventions. It is critical to improve our understanding of drivers of weight loss responsiveness as this may help identify causal factors in obesity and may ultimately lead to new or more personalized prevention or treatment opportunities. This study will examine how genetics, gut microbiota (GM), and metabolites contribute towards measures of responsiveness during an ongoing 1-year lifestyle weight loss trial of the standard weight loss approach, daily caloric restriction (DCR), versus a novel alternative, intermittent fasting (IMF). This understanding of the causal relationships between omic profiles and weight loss/health improvements in an interventional setting will target an investigation into GM and metabolites as mediators of the association between genetic obesity risk and obesity-related phenotypes in larger epidemiological cohorts. Preliminary data from a participant subset in the intervention that provides evidence of feasibility and informs the hypotheses for the following innovative aims: Aim 1: Assess longitudinal changes in GM and metabolites, as well as the predominant drivers of these changes, during a lifestyle weight loss intervention of DCR versus IMF. Aim 2: Evaluate associations of a measure of genetic risk for obesity and GM/metabolites with responsiveness during a lifestyle weight loss intervention. Aim 3: Investigate whether GM/metabolites mediate the association between a multiethnic obesity polygenic risk score and measures of obesity in two epidemiological cohorts. This Career Development Award will also provide the Candidate with the opportunity to gain important skills and expertise. She has a clear plan to gain training related to: 1) processing shotgun metagenomic GM and metabolomic data; 2) causal inference and other relevant methods for the analysis of multi-omics data; and 3) multiethnic polygenic risk scores. The training plan involves coursework, conference attendance, and in-person training with leading scholars, in conjunction with guidance from a mentorship team of renowned leaders in their respective fields of genetic epidemiology (Dr. Leslie Lange; Primary Mentor), bioinformatics and the human gut microbiome (Dr. Catherine Lozupone), biostatistics (Dr. Katerina Kechris), and clinical weight loss interventions (Dr. Victoria Catenacci). This training plan combines the exceptional research environment at the University of Colorado Anschutz Medical Campus with generous institutional support and ample resources, as well as an outstanding network of advisors. This work will facilitate the candidate's successful transition to an independent career in molecular epidemiology with the expertise to integrate complex types of omics data in order to improve our understanding of disease pathophysiology and to inform innovative prevention and intervention efforts for obesity and CVD.

NIH Research Projects · FY 2025 · 2022-02

Enrollment in high deductible health plans (HDHPs) has increased 50% in the past 5 years, with more than 45% of privately-insured Americans enrolled in such plans. HDHPs require patients to pay 100% of the costs of most care (excluding some primary or secondary preventive care) out of pocket until a (high) deductible is met. Services provided for chronic disease management are subject to this deductible. Therefore, it is unclear whether this form of insurance supports healthy aging, especially for the 70% of Americans aged 50-64 (an NIA priority population) with chronic illness. High patient cost-sharing has caused decreases in use of both necessary and unnecessary health care. However, it is not known whether reductions in healthcare use mean that patients are not receiving the types of healthcare required to manage their chronic diseases. The overarching goal of this study is to ascertain whether the high cost-sharing design of HDHPs results in chronically-ill patients not receiving the evidence-based, high-quality care recommended for management their illnesses and secondarily, whether the cost barriers built into their design result in poorer health outcomes for HDHP enrollees. We also estimate the magnitude of patient spending required to receive high quality care in HDHPs versus non-HDHPs and investigate any race-, gender-, or income-based disparities in care experienced by HDHP patients. We focus our HDHP investigations on patients who have one or more of the following common chronic illnesses: diabetes, coronary artery disease, heart failure, asthma, hypertension, or major depression. Lastly, we use a temporary benefit design change enacted during the COVID-19 pandemic as an exogenous shock to understand the impact of HDHPs on receipt of recommended medical care. Previously HDHP research has not focused on impacts on populations most in need of services (i.e., those with chronic conditions) and the impact of HDHPs on disparities. We evaluate, for the substantial proportion of mid-life adults with high healthcare needs, whether this common benefit design results in lower utilization of care (due to high patient out-of-pocket costs) and adverse health outcomes. Results from this work are policy-relevant. Findings can inform regulations regarding the exemption of chronic disease management from insurance deductibles and identify the best way to support value-based insurance design. To ensure findings reach decision makers, we will disseminate results through RAND Research Briefs to insurance purchasers, leaders at insurance companies, state departments of insurance, and members of Congress, in addition to traditional mechanisms of dissemination to the scientific community.

NIH Research Projects · FY 2026 · 2022-02

Abstract Early events in HIV-1 lifecycle, such as post-fusion trafficking of viral cores across the cytoplasm, through the nuclear pore complex (NPC) and into the nucleus, remain poorly understood due to limited information about virus-host interactions. Interactions of the core’s surface, which is composed of the capsid protein (CA) arranged into large hexameric lattices and exactly 12 pentamers, with a variety of host cell proteins that aid infection (dependency factors) are crucial for this journey. However, their full identity and molecular mechanisms of action remain largely unknown. Our preliminary studies resulted in the following two principal discoveries. 1) We have identified SEC24C as a new, crucial HIV-1 host dependency factor. SEC24C is a predominantly cytoplasmic protein that employs a phenylalanine-glycine (FG)-motif to specifically interact with the hexameric CA lattice at the hydrophobic pocket comprised of two adjoining subunits. These novel findings, coupled with the known roles of other CA-binding FG-motif containing cellular factors NUP153 and CPSF6 in nuclear import and integration site selection of HIV-1, suggest that these proteins collectively provide a dependable platform for continuous HIV-1 journey throughout different cellular compartments during the virus ingress to ensure productive infection. 2) We discovered a prion- like domain (PrLD)-mediated mechanism for avid binding of SEC24C, NUP153 and CPSF6 to hexameric CA lattices. Our preliminary cryo-EM and HDX-MS studies provide novel structural information indicating that, in addition to known CPSF6 FG peptide binding to the cognate CA hydrophobic pocket, PrLD-PrLD interactions enable polyvalent assembly of CPSF6 molecules along the extended lattices of adjoining CA hexamers. Our virology experiments further support an essential role of CPSF6 PrLD in functional virus- host interactions in infected cells. To extend these exciting, paradigm-shifting preliminary studies we propose the following three specific aims: Aim 1 will define a role of SEC24C in HIV-1 infection; Aim 2 will elucidate interplay between SEC24C and other CA-binding host factors; Aim 3 will determine the structural basis for avid recognition of hexameric HIV-1 CA lattices by FG-motif and PrLD containing cellular factors SEC24C, NUP153 and CPSF6. To accomplish these aims we have assembled a highly collaborative team with complementary expertise in virology, proteomics, biochemistry and X-ray crystallography (Kvaratskhelia), cryo-EM (Asturias), live cell microscopy (Melikian) and HDX-MS (Griffin). These studies are expected to uncover novel virus-host interaction mechanisms crucial for HIV-1 infection. Furthermore, elucidating structural determinants for functionally relevant interactions of SEC24C, NUP153 and CPSF6 to hexameric CA lattices will provide a new frontier in HIV-1 structural biology and improve our understanding of these virus-host interactions as an important therapeutic target.

NIH Research Projects · FY 2026 · 2022-02

The goal of the University of Colorado Anschutz Medical Campus (UC AMC) Preparation in Interdisciplinary Knowledge to Excel (PIKE) PREP is to offer a multi-dimensional mentoring and research training experience to inspire URM postbaccalaureate students to enroll and succeed in a PhD or MD-PhD program and commit to a career in biomedical research. PIKE PREP will address barriers URM students are known to face in entering and completing graduate programs. The PIKE PREP is a structured, yet highly customizable program that offers a variety of activities that address the individual Scholar's research weaknesses as well as identify and reinforce their strengths. We will couple evidence-based practices with metric-based evaluations to gauge and improve efficacy of activities. The PIKE will focus on key attributes which we and others have identified as necessary for graduate school success, developing these attributes through programmatic and mentorship activities, including mentorship in Research, Mental Health, Career, Leadership, and Role Model. Our main purpose is to advance at least 80% of PREP Scholars to PhD or MD-PhD biomedical research programs on our campus or at another top-tier PhD program. Aim 1 is to engage the Scholars in research while building their technical skills through an exceptional biomedical research-focused experience. This aim will focus on providing Scholars in an in-depth research experience by taking advantage of a wide range of resources as well as highly productive and dedicated mentors with research expertise across a wide spectrum of biomedical-related research. Aim 2 is to improve the Scholar's competitiveness for PhD programs by providing analytical skills enhancement and substantial guidance on the graduate school application process. Activities in this aim will help build a strong foundation in scientific reasoning, rigorous research design and data analysis and interpretation. We will also guide students to prepare exceptional graduate school applications, refine interview skills, and lead them to the most appropriate graduate programs that match their interests and needs. Aim 3 will provide an assortment of high-quality professional and career development activities to complement each PREP Scholar's research experience. We work closely with the Scholars and mentors to develop, monitor, and refine IDPs for each Scholar based on individual career needs. Routine evaluations of IDPs by our PREP Leadership Team will ensure that the PREP Scholar is making excellent progress and that the training objectives of the IDP are met. Aim 4 will provide a continuous and dynamic mentoring and community-building research-centered experience. PREP Scholars will have various opportunities to network with URM and non-URM mentors, graduate students, postdocs and faculty and different forums to become integrated into the social, research, and academic environments. The successful implementation of these programmatic initiatives will increase the number of highly qualified and competitive URM students that enter and succeed in a PhD program.

NIH Research Projects · FY 2026 · 2022-02

Project Summary/Abstract The production of messenger RNA (mRNA) is the primary event in gene expression where genetic information is transfered from the gene’s DNA into a disposable RNA copy. Corruption of this process is a hallmark of many diseases including cancer. mRNA synthesis requires not only synthesis of an RNA transcript but maturation of that transcript by 5’ capping, excision of introns and splicing of exons and 3’ end formation by cleavage and polyA tail addition. The mRNA processing steps that radically transform the primary transcript occur largely co-transcriptionally; that is to say the substrate of mRNA processing is the growing nascent RNA that is extruded by an RNA polymerase II (pol II) molecule at rates of 500-5000 bases/min. Our working model is that synthesis and processing of a mRNA precursor are carried out in an integrated fashion within a dynamic 'mRNA factory' complex that includes both RNA polymerase and processing factors some of which make direct contacts with the pol II C-terminal domain (CTD). The goal of our work is to understand how growth of the RNA chain by transcription is coordinated with its folding into RNA secondary structures, its association with RNA binding proteins, and its maturation by splicing and 3’ end formation. These important features of nascent RNA metabolism are all affected by how fast the RNA chain grows. Therefore it is important to discover how the speed of pol II is controlled as it travels along genes. When pol II completes its journey to the end of the gene, the highly stable transcription elongation complex must be actively disassembled to recycle the enzyme and prevent it from invading neighboring genes. We will investigate how the process of transcription termination is achieved in carefully controlled ways at the 3’ ends of genes and also within genes where termination can occur “prematurely”. We will use genetic and genomic approaches in human cells to investigate these three Key Challenges: I. What is the relation between pre-mRNA processing, nascent RNA folding, RNA binding protein (RBP) binding, and transcription elongation? II. How is the speed of transcription elongation regulated? III. What mechanisms terminate pol II transcription within genes and downstream of genes?

NIH Research Projects · FY 2025 · 2022-02

PROJECT SUMMARY/ ABSTRACT Hemolytic syndromes including sickle cell disease (SCD) are devastating illnesses that affect over 100,000 people in the United States. Each of these patients suffers a broad spectrum of cardiopulmonary complications and exercise intolerance caused by red blood cell hemolysis, high plasma levels of cell free hemoglobin (Hb), endothelial cell dysfunction, and tissue hypoxia. Pulmonary vascular disease in the form of pulmonary hypertension, is significantly increased in this population, without any adequate treatment options. This grant application and projects herein focus on elucidating the mechanistic un- derpinnings of macrophage activation and their contribution to the progression of SCD PH. Our data shows a unique macrophage (Mϕ) phenotype occurs in both deceased adult SCD patients di- agnosed with PH as well rodent models of SCD PH. This phenotype is described by intracellular accu- mulation of Hb, expression of HO-1, and mitogenic, inflammatory and vasoconstrictor mediators that are also associated with hypoxia (HX) induced PH. Furthermore, like hypoxia driven PH, SCD patient lung macrophages accumulate in the pulmonary adventitial regions surrounding remodeled pre-capillary pulmonary arterioles that show plexiform lesions and re-canalization of small pulmonary arterioles. The phenotypic similarities between rodent models and human SCD with PH indicate a novel maladaptive immune response to concomitant bouts of Hb and HX exposure. Moreover, by knocking out circulating mϕs in a rat model of group 5 PH, the response to combined Hb and hypobaric HX was significantly at- tenuated in rats, suggesting a critical role for mϕs in the exacerbation of SCD PH. We hypotheses that persistent bouts of hypoxia-induced erythrocyte sickling are a critical process that drives Mϕ removal of damaged RBCs causing accumulation of Mϕ iron, loss of Mϕ iron homeostasis and progressive SCD-PH. We further hypothesize that impaired iron homeostasis facilitates in- tracellular oxidation and exposes the local pulmonary vascular micro-environments to labile iron mediated oxidation and accelerated lung peripheral vascular remodeling. To test this hypoth- esis we propose the following specific aims. Aim 1 will determine how circulating monocytes with a high iron content are metabolically reprogramed in patients with differing severity of SCD. Aim 2 in vivo and in vitro will determine the contribution and mechanism by which iron loaded Mϕ contribute to SCD; and Aim 3 will test the effectiveness of trans- ferrin, ferroportin inhibitor, and iron chelators alone or in combination as a therapeutic intervention to halt Mϕ contributions to SCD PH. An in-depth understanding of these relationships will allow us to identify new therapeutic targets to pulmonary hypertension concomitant with SCD.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY Epigenetic gene repression by heterochromatin is necessary for multi-cellular organismal development and genome stability. Mis-regulation of heterochromatin is a cause of multiple diseases through perturbed gene expression patterns. The long-term goals of our research program are to determine at a molecular level how noncoding RNAs (ncRNAs) participate in heterochromatin formation and function. Many ncRNAs act in the nucleus to regulate gene expression through association with chromatin regulatory machinery. We and others have shown that certain RNA-mediated heterochromatin pathways require intermolecular RNA-RNA interactions between ncRNAs and nascent RNA that serve as the trigger to form heterochromatin. However, many systems where RNA is implicated have unexplored molecular mechanisms of action. We will address three high-level major outstanding questions in the field: 1) Which heterochromatin systems are controlled through RNA-RNA interactions? 2) How is heterochromatin built around nascent RNA? 3) How are RNA binding proteins re- purposed from mRNA processing functions to contribute to RNA-mediated heterochromatin formation? Our research program focuses on multiple heterochromatin systems that incorporate different species of noncoding RNAs. We study long noncoding RNAs (lncRNAs), such as HOTAIR and pericentromeric transcripts, which can inhibit heterochromatic histone modifiers to control their activity. In addition, we have developed the first biochemical system to study the human nuclear piRNA pathway, which uses base-pairing of the small piRNA to target nascent transcripts of repeats that then are suppressed via heterochromatin. Finally, we address how RBPs such as the N6-methyladenosine reader YTHDC1 can work with ncRNAs to promote gene repression. We use biochemical, structural, cell biological, and genomic approaches to study these models of RNA-regulated heterochromatin. Mechanistic insight into these pathways will provide a fuller understanding of how they work normally and in disease, which will prove useful in targeting for pharmacological intervention. Relevance to public health Noncoding RNAs are produced from regions of the human genome originally thought to be "junk" DNA. Many ncRNAs participate in epigenetic mechanisms of gene regulation and mis-regulation can lead to diseases such as cancer. ncRNAs are therefore clear candidates to provide a missing link to understanding the molecular mechanisms of many human diseases for which there is a "hidden heritability" factor that has not yet been identified.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY Acute asthma exacerbations are the primary cause of morbidity and mortality in children with asthma. Current treatment for acute asthma exacerbations in the pediatric Emergency Department (ED) follows an one- size-fits all approach including inhaled bronchodilators and systemic corticosteroids. However, treatment response to initial protocolized therapies is variable and unpredictable presenting a significant management challenge for ED clinicians. Unfortunately, the pathobiologic mechanisms driving treatment response remain unclear, and an effective method to predict treatment response does not exist. Thus, ED clinicians frequently struggle with treatment and disposition decisions leading to over-utilization of therapies, prolonged ED length of stay, and hospitalizations in responders, and delays in appropriate therapy for non-responders. The extensive variation and inefficiency in care highlights the critical need for tools to inform more precise and effective ED management strategies for acute asthma exacerbations. The nasal transcriptome and airway oscillometry (AOS) are novel biologic and physiologic markers with strong potential to address this unmet knowledge and practice gap. This proposal aims to apply an innovative biomarker-directed, individualized approach to ED asthma management by leveraging these novel markers to pursue the following specific aims: 1) determine the utility of AOS as an objective measure of ED treatment responsiveness; 2) identify airway endotypes of ED treatment responsiveness using nasal transcriptomics, and 3) derive and internally validate a clinical prediction rule incorporating biologic and physiologic markers to determine ED treatment responsiveness in children with acute asthma exacerbations. To achieve these aims, the candidate, Nidhya Navanandan, MD, will leverage an existing study infrastructure for enrolling children with acute asthma exacerbations in the ED, developed in conjunction with her mentors during her institutional career development award. As a pediatric emergency medicine physician, Dr. Navanandan is uniquely positioned to accomplish the proposed K23 research and training aims. Her long-term goal is to become an expert in clinical and translational research methods to improve the effectiveness of emergency care for pediatric asthma. Dr. Navanandan has developed a detailed career development plan consisting of mentorship, didactic coursework, and hands-on laboratory and research conduct experience in order to expand her knowledge and skills in leadership of prospective studies, discovery and application of novel markers for clinical practice, and predictive analytics. Dr. Navanandan has assembled a multidisciplinary team of mentors with extensive clinical and translational research experience and topical expertise in the above realms to ensure her success in achieving the stated specific aims and career goals. This proposal will allow Dr. Navanandan to transition to an independent physician-scientist and prepare her for future R01-funding.

NIH Research Projects · FY 2026 · 2022-01

ABSTRACT The Colorado Summer Institute for Biostatistics (CoSIBS) has extended the reach of the SIBS network into the western US. In its first five years 26/77 (34%) of participants came from western states. In keeping with program objectives CoSIBS will continue to inspire the next generation of biostatisticians and data scientists to pursue advanced study and research careers in the biomedical, behavioral, and quantitative health sciences. We will recruit an enthusiastic and diverse group of advanced undergraduate and early-stage graduate students who are interested in exploring the frontiers of data sciences in the development and application of new methods for using big data to guide the diagnosis, care, and prevention of disease. The proposal is structured to provide innovative educational programs using the established research in biomedical data sciences with NHLBI and NIAID-related research programs that cut across departments at University of Colorado Denver and affiliated institutions. The proposal draws together a range of faculty including faculty from the Department of Biostatistics and Informatics, the Department of Mathematical and Statistical Sciences, and faculty from other departments with research interests in bioinformatics and data science. CoSIBS will be a 6-week resident summer institute for approximately 20 advanced undergraduate students or recent graduates. The institute will consist of approximately 4 hours per day of class and laboratory instruction. The didactic portion will be at a standard that would be typical of a one-year course in basic statistics as often required in undergraduate curricula. In addition, students will have mentored projects that are tied to ongoing research groups at UCD and affiliated institutions. Projects will be co-mentored by faculty with research interests in biostatistics, statistical genetics, or bioinformatics, and a companion biomedical investigator from a leading NHLBI-related research group at UCD. The projects will reinforce course content in study design, data analysis, software applications, and the preparation of results for publication or presentation to biomedical researchers. An innovative mentored summer fellowship opportunity will be launched with an institutionally supported stipend for students from historically underrepresented populations to work with DBI faculty on collaborative research projects during the weeks preceding and following the 6-week CoSIBS program.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY/ABSTRACT Human Papillomaviruses (HPV) infect >90% of the population and cause >30,000 cervical, anogenital, and oropharyngeal cancers annually. HPV vaccines have been available in the US since 2006 but only 54% of 11- 12 years-olds are vaccinated - well below national goals of 80%. As the title of the PAR to which this applica- tion responds suggests (PAR-19-360), “Linking the Provider Recommendation to Adolescent HPV Vaccine Up- take” is a key strategy for increasing adolescent HPV vaccination. Many studies show that a primary driver of low adolescent HPV vaccination is a poor quality HPV vaccine recommendation from providers. While many interventions to address this have been developed, few have been found in rigorous trials to increase vaccina- tion and also be feasible to implement in busy clinical settings. An exception is the Physician Communication, or PCOM, intervention developed by our group. PCOM focuses on teaching providers to use a 2-step verbal communication process for their recommendation:1) start the vaccine discussion using a “presumptive” format (i.e. “Let's get the HPV vaccine done today”) and 2) use motivational interviewing (MI) techniques to address parental vaccine hesitancy, if needed. In a large, cluster-randomized trial, PCOM increased adolescent HPV vaccine initiation rates among 11-12 year olds by 8 percentage points (PP) more than controls– a relatively large effect size for a vaccination intervention. Limiting broad dissemination of PCOM is the significant level of research team facilitation needed to teach practices how to use the PCOM components. To address this, we propose to develop a “Virtual” version of the PCOM intervention (“PCOM-Virtual”) and test it for non-inferiority to the original PCOM intervention (“PCOM-Standard”) for increasing adolescent HPV vaccination. By using Dissemination & Implementation (D&I) science principles throughout the process and collecting information on how contextual practice, provider and patient factors influence PCOM use, we anticipate creating a “shelf ready” version of PCOM with an associated “User Manual” to foster dissemination. Our Aims are to: 1. Develop the “PCOM-Virtual” intervention using principles of D&I Science from existing prototypes. 2. Compare the efficacy of “PCOM-Virtual” vs. “PCOM-Standard” in improving adolescent HPV vac- cine utilization. A cluster-randomized non-inferiority trial will be conducted in 30 primary care practices in Kansas – a locale with some of the lowest adolescent HPV vaccination rates nationally. 3. Examine whether practice, patient and provider characteristics are associated with variability in the efficacy of PCOM-Virtual and PCOM-Standard. Mixed methods and the PRISM framework will facili- tate understanding how context influences the interventions' implementation to inform a User Manual. By creating an easily implementable version of the PCOM intervention, and disseminating it widely we believe we can have a substantial impact on adolescent HPV vaccination levels.

NIH Research Projects · FY 2025 · 2022-01

ABSTRACT Food insecurity is known to be associated with metabolic and cardiovascular disease in adults. However, there is a gap in our knowledge; specifically a lack of studies investigating factors that influence the emergence of cardiometabolic risks from food insecurity during childhood and adolescence. An improved understanding of the role of timing and severity of food insecurity on children’s cardiometabolic health (CMH), the mechanisms/pathways between food insecurity and CMH, and the factors that buffer against or exacerbate this risk are needed to inform intervention development to prevent later cardiometabolic disease amongst children who are food insecure. The main objective of this prospective study is to understand how cardiometabolic risk arises from food insecurity during childhood and identify malleable factors for intervention to buffer the negative health impact of food insecurity on adolescent’s CMH. We will leverage an established cohort of racially/ethnically diverse children from low-income households (n=627) participating in a prospective, longitudinal study examining childhood obesity disparities (HL126171: Berge PI). Survey and ecological momentary assessment (EMA) data have already been collected, including measures of food insecurity, household environment (e.g., food environment, family functioning), child health behavior (e.g., diet quality, sleep, physical activity) and parent behaviors (e.g., feeding practices, stress, coping) at two time points (~24 months apart) at ages 5-10. New measures proposed at ages 11-16 will include: (1) measures of CMH (Life’s Simple 7 and state-of-the-science measures of body composition, metabolic and cardiovascular function, oxidative stress and inflammation (i.e., cytokines)), (2) Geographical Information Services (GIS) data, and (3) focus groups with families, school representatives, community organizations, and healthcare providers to identify intervention targets and refine policies to reduce harmful health effects of food insecurity. Specifically, this state-of-the art study will: (1) determine the impact of exposure to household food insecurity (i.e., severity, timing) on child/adolescent CMH; (2) identify mechanisms/pathways between food insecurity and CMH; (3) evaluate the extent to which child behaviors, parent factors, and access to resources impact the relationship between food insecurity and CMH; and (4) using mixed-methodologies (biomarkers, focus groups, EMA, GIS), prospectively examine the dynamics (e.g., severity, timing) of household food insecurity throughout childhood and adolescence, and identify risk and protective factors at the level of the family, neighborhood, school, and community to inform intervention and policy development. This approach is innovative because it represents a departure from the status quo by focusing on when and how cardiometabolic risk emerges from food insecurity during childhood and the multi-level factors which buffer against or exacerbate this risk, and significant because it is expected to have implications for intervention development to prevent future cardiometabolic diseases among adolescents from food insecure households.