University Of Colorado Denver

NIH Research Projects · FY 2026 · 2022-08

Complications that arise secondary to an exaggerated innate immune response, such as multiple organ failure, are a major cause of late-stage mortality in trauma patients. My overall goal is to initiate an innovative and translational research program focused on elucidating mechanisms through which the vascular endothelium regulates the host inflammatory response to severe trauma. In particular, my research is focused on the immunomodulatory functions of the antithrombin (AT)-heparan sulfate system. AT elicits anti-inflammatory signaling upon binding to specific heparan sulfate proteoglycan (HSPG) receptors on the endothelial surface that contain a 3-0-sulfate (3-0S) modification. Our ongoing experiments demonstrate that dysregulation of the ATHSPG system is a novel mechanism driving inflammation and organ injury following severe trauma and hemorrhagic shock. However, the mechanisms that govern 3-0S HSPG expression and AT binding following trauma is a major knowledge gap in the field. Understanding these mechanisms will enable us to develop novel clinical tools to attenuate aberrant inflammation following trauma and treat or prevent subsequent organ failure. The next 5 years of my proposed research program will focus on 3 developing programmatic areas that seek to elucidate 1) mechanisms that mediate 3-0S HSPG degradation; 2) mechanisms that regulate 3-0S HSPG biosynthesis; and 3) the biological role and therapeutic potential of unique AT variants capable of regulating inflammation when 3-0S HSPG expression is reduced. Results of these investigations have broad-reaching implications for many conditions in which inflammation contributes to the pathogenesis, such as sepsis, transplantation, and COVID-19. Funding from this R35 award will 1) enable the establishment of my highly innovative, long-term research program that is guided by the NIGMS mission; 2) advance our basic understanding of the host inflammatory response to trauma; and 3) create novel therapeutics to improve longterm survival and quality of life for the critically ill.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY / ABSTRACT Spinal cord injury (SCI) is a devastating event in terms of a person’s health, physical function, costs (direct and indirect), and life expectancy. It has been conventionally thought that individuals with severe SCIs, with no motor function below the level of injury, will not recover the ability to functionally move their lower extremities or voluntarily walk. However, spinal cord stimulation (SCS) has emerged as a promising intervention challenging this long-held assumption. While spinal cord stimulation allows for restored voluntary movement after severe SCI, there is a lack of mechanistic understanding regarding how it works and why some individuals respond better than others. Thus, the overall objective of this proposal is to apply neuroimaging biomarkers to generate fundamental knowledge regarding responsiveness to spinal cord stimulation (SCS). Aim 1: Use neuroimaging biomarkers to understand responsiveness to epidural SCS in participants with severe SCI, during volitional movement and standing tasks. Using high-resolution MRI in a prospective design, the applicant hypothesizes that the laterality of cord damage, detected prior to surgical implantation, will predict ipsilateral lower extremity muscle responsiveness to epidural SCS prior to any training. Aim 2: Use neuroimaging biomarkers to understand responsiveness to transcutaneous SCS in participants with severe SCI, during volitional movement tasks and sensory examination. Using high- resolution MRI in a prospective design, the applicant hypothesizes that total spinal cord spared tissue will predict bilateral lower extremity muscle responsiveness to transcutaneous SCS prior to any training, and that posterior cord spared tissue will predict light touch sensory recovery prior to any training. Significance: Successful completion of these Aims will advance the NIH/NICHD NCMRR aim: “to enhance the health, productivity, independence, and quality of life of people with physical disabilities.” One important problem in the field of SCS is a lack of foundational knowledge on why the intervention works. Neuroimaging holds pronounced potential to address this problem. Neuroimaging biomarkers will not only improve the understanding of responsiveness to this intervention after SCI, but will also help drive individualized approaches for using SCS, selection of epidural versus transcutaneous SCS, prognosis for improvement using SCS, and the identification of who is likely to optimally respond before activity-based recovery training. Completion of the proposed aims will lead to the high likelihood of sustained, powerful influence on the SCS field, laying a vital foundation for using MRI biomarkers to guide SCS intervention, ultimately improving the clinical management of persons with SCI.

NIH Research Projects · FY 2025 · 2022-08

Project Abstract Glioblastoma (GBM) is the most devastating and aggressive brain tumor in adults, with patients surviving a median of only 14.6 months. Hidden behind the blood-brain and blood tumor-barrier (BBTB), the glioma cells migrating away from the tumor and invading surrounding brain tissue, are the source of recurrence. The invasive cells are not readily accessible to most drug therapeutics, and targeting these cells is an essential goal for achieving better treatment outcomes. Nanoparticles hold promise for drug delivery, but their penetration of the BBTB is limited, and the efficiency of targeting invasive cells remains unknown. We recently discovered that fluorescent indocarbocyanine lipids (ICLs) formulated in PEGylated Lipid Nanoparticles (PLNs) exhibit highly efficient glioma extravasation, with a single injection resulting in accumulation in ~60% of tumor cells and up to 30% of injected dose per gram of brain tumor. Furthermore, data in highly invasive models demonstrate PLNs reach invasive cells at the tumor/brain margin. These findings offer a unique opportunity to comprehensively understand the mechanism of accumulation of lipid nanoparticles and improve drug delivery to invasive gliomas. We will pursue the following specific aims: 1) Study the trafficking mechanism of ICLs across the BBTB and in tumors. In this aim, we will test the hypothesis that lipids migrate in tumors via extracellular vesicles; 2) Understand the role of lipid structure and formulation in targeting glioma cells and the tumor immune microenvironment. This aim will test if accumulation in invasive cells and immunosuppressive cells can be further improved through lipid chemistry, formulation and targeting to glioma marker IL13R2; 3) Exploit ICLs to understand and improve small molecule delivery to invasive gliomas. We will explore our previously developed chemistry to conjugate small molecules to ICLs to improve their delivery, drug release, and therapeutic efficacy of cyclin-dependent kinase inhibitor dinaciclib in invasive mouse and patient-derived glioma models. These studies will expand our understanding of the drug delivery process and guide treatment of invasive gliomas with brain tumor-penetrating nanomedicine.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Cisplatin (Cis)-based Neoadjuvant chemotherapy (NAC) is the standard of care prior to cystectomy, for patients with muscle-invasive bladder cancer (MIBC). Up to 30% of patients respond and show no residual tumor at cystectomy with >80% survival, but “non responders” have <30% chance of surviving 5 years. Thus, improving the effectiveness of Cis-based NAC will greatly improve outcomes in BC. Through whole-genome CRISPR-Cas9 synthetic lethal screens in Cis-resistant human BC cell lines, we discovered NPEPPS as a novel and druggable target whose expression determines sensitivity to Cis. NPEPPS was the only one of 13 M1 aminopeptidases found to be synthetic lethal with Cis. Depletion of NPEPPS enhanced Cis therapy and reduced growth in animal models. To find how NPEPPS drives these two phenotypes, we used mass spectrometry (MS) to identify the proteins that are in complex with NPEPPS in BC cells. We found NPEPPS in complex with subunits (LRRC8A- E) of the volume regulated anion channel (VRAC), a recently identified mechanism of platinum (Pt) cellular import. In BC cells, LRRC8A/D depletion increases resistance to both cisplatin and carboplatin, while NPEPPS depletion had the opposite effect. Supporting a role in human BC growth, NPEPPS expression is associated with poor patient outcome regardless of chemotherapy use. Leveraging our MS results, we developed an approach to prioritize candidate genes found in complex with NPEPPS that most likely affect growth, are associated with aggressive disease, and are prognostic markers. Thus, we propose the Guiding Hypothesis that NPEPPS drives Pt resistance and tumor growth in BC by inhibiting VRAC activity and interacting with genes regulating cell proliferation respectively. Specific Aims test this hypothesis with the Objective to lay the foundation for novel approaches to improve the outcomes for BC patients. In Aim 1 we will test the hypothesis that NPEPPS aminopeptidase activity is required for Pt resistance and growth using enzymatically dead NPEPPS mutants in vitro and in vivo. Next, we evaluate our top candidate gene CHD2, a chromatin regulator and putative tumor suppressor for its role in NPEPPS-driven tumor growth and the dependency of this role on NPEPPS enzymatic activity. In Aim 2 we will determine the role of LRRC8A/D in NPEPPS-mediated Pt resistance. We have used molecular modeling to identify residues on NPEPPS that interact with LRRC8A/D. Site-directed mutagenesis of these residues will test the hypothesis that direct interaction of NPEPPS with LRRC8A/D reduces the ability of VRACs to properly function and contributes to NPEPPS-mediated Pt resistance in vitro and in vivo. To establish the preclinical rationale for the effect of novel, anti-neoplastic agents that circumvent Pt resistance, Aim 3 will test the hypothesis that Tosedostat (Tose), a clinically well-tolerated aminopeptidase inhibitor, enhances the sensitivity of BC to Pt, and NPEPPS expression is required for this. The impact of Tose on BC cells ± NPEPPS expression will be examined for growth, Pt sensitivity and import in vivo in human BC models of localized and metastatic disease. Organoids derived from patient tumors before Cis-based NAC, will be tested with Pt ± Tose.

NIH Research Projects · FY 2026 · 2022-08

PROJECT SUMMARY This is a resubmission application for a research education program to support individuals from diverse backgrounds to pursue the study of bioengineering and a career in research. The overall objective is to recruit talented students from the Community College of Denver, a minority serving institution (MSI), into the undergraduate Bioengineering program at the University of Colorado Denver, also a MSI, and to provide these students with the skills necessary for postgraduate studies in bioengineering, biomedical sciences, and medicine. The rationale for the proposed research education program is that community college transfers are an important, but often neglected, part of the pipeline of students with diverse backgrounds that enter undergraduate bioengineering programs. In order to support this group of students we will pursue the following specific aims: (1) a pre-transfer advising program on academic, cultural, and financial issues to make for a successful transfer to a four-year degree program, (2) a summer bridge program in which students earn credit towards their degree and learn essential skills in design thinking and computational thinking while being oriented into campus culture, (3) academic year activities that includes paid engineering apprenticeships and a layered mentoring approach, (4) a summer research experience in bioengineering at an academic medical center that includes skills workshops and responsible conduct in research training, and (5) an honors and leadership program that provides additional research experience and leadership training in the junior and senior years as well as entry into a combined B.S./M.S. program. This approach is innovative because it targets a unique population of students with diverse backgrounds and will yield an educational research program that caters to their strengths and needs. The proposed education program is significant because it will provide both quantitative and qualitative data on how these approaches support science identity, academic efficacy, scholarship, and matriculation into postgraduate programs. If successful, this program could be expanded at CU Denver and replicated at other institutions to support community college transfers in bioengineering and related fields.

NIH Research Projects · FY 2025 · 2022-08

Project Summary Synaptic inhibition in the brain is mostly mediated by GABAergic inhibitory synapses, which are essential in controlling neuronal firing, neuron and circuit excitability, and synaptic plasticity. Inhibitory synapses undergo multiple modes of plasticity but the molecular mechanisms that drive persistent inhibitory post-synaptic plasticity remain poorly defined. As translation of synaptic proteins is essential for many forms of plasticity, synthesis of new inhibitory post-synaptic proteins likely plays a key role in sustaining changes in inhibitory synaptic strength. However, little is known about which inhibitory synaptic proteins are translated, where they are produced and how this process is regulated during activity. Our preliminary work suggests that multiple inhibitory synaptic proteins are locally translated in neuronal dendrites during inhibitory long-term potentiation. This process is tightly regulated by miRNAs, potent negative-regulators of translation. This proposal will examine the control of inhibitory synaptic plasticity by local miRNA-dependent translational regulation, using in vivo and in vitro advanced imaging and electrophysiological approaches. The proposed studies are significant as they have the potential to reveal key mechanisms that drive persistent and long-lasting changes in synaptic inhibition. Given the important roles of inhibitory synapses in the brain, this project will provide crucial insight into the underlying mechanisms of learning, memory and cognition.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY / ABSTRACT There is substantial interest in identifying intricate mechanisms of the morphine-mediated effects and opioid addiction in order to expand strategies for novel therapies. Although the role of nonspecific thalamus in regulation of consciousness is well documented, its possible role in the morphine-mediated effects and addiction is not well studied. Principal Investigator (PI) has recently discovered the role of CaV3.1 isoform of T-type calcium channels (T-channels) in the central medial nucleus of thalamus (CeM) in neuronal excitability regulation and thalamocortical oscillations during anesthesia. Furthermore, based on the new exciting preliminary data, the overarching goal of this proposal is to determine the functional role of CaV3.1 isoform of thalamic T-channels in the morphine effects and addiction. It is hoped that this may form foundation for novel approaches to treatment and/or prevention of opioid overuse. In Aim 1, I will use patch-clamp recordings and 2-photon calcium imaging from acute brain slices ex vivo and projection-specific targeting and manipulating CeM neurons to determine acute and lasting effects of morphine exposure (after repeated morphine exposure and morphine-induced conditioned place preference – CPP) on T-channels and excitability of CeM neurons. In Aim 2, I will use behavioral experiments relevant to the addiction: CPP and self-administartion (SA) models, in vivo viral silencing technologies (with small hairpin RNA (shRNA) and Cre-specific knocking-out (KO)) and chemogenetic method for transient behavioral modulation to investigate functional role of thalamic T-channels in addiction. Completion of these aims will generate new insights into the mechanisms of the morphine effects and opioid addiction that could identify new treatment options. The PI will receive mentorship and technical training in addictive behavior and viral technologies by experts in electrophysiology and viral technologies in motivated behaviors. University of Colorado and the Department of Anesthesiology provide exceptional facilities and resources for completing the proposed experiments, as well as having an exceptional reputation and record of accomplishment for mentoring and transitioning early-stage into independent investigators. The proposed training, education and research will provide the PI with the technical and professional training to become a successful, independent investigator.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Learning, cognition, and memory require dynamic remodeling of hippocampal synapses, which in turn requires Ca2+/calmodulin-dependent kinase II (CaMKII). CaMKII mediates two opposing modes of synaptic plasticity, long term potentiation (LTP) and depression (LTD), that are induced by distinct Ca2+ stimuli. Both low and high Ca2+ induce CaMKII autophosphorylation (p) at T286, that is required for both LTP and LTD. Additionally, LTP requires CaMKII binding to the NMDA receptor subunit, GluN2B, during high [Ca2+] while LTD requires CaMKII autophosphorylation at T305/306 during low [Ca2+]. Further, these three mechanisms can undergo complex cross-regulation which requires the CaMKII 12-meric holoenzyme. Interestingly, pT286 positively regulates both GluN2B binding and pT305/306 while GluN2B binding and pT305/306 are mutually exclusive. It is unknown how these reactions and interactions are spatiotemporally encoded within holoenzymes and thus how LTP versus LTD signal computation is accomplished by CaMKII. For example, it has been shown that pT286 must occur between two neighboring kinase domains in the holoenzyme. It is still unclear what determines a functional kinase domain neighbor. Moreover, in vitro binding studies have shown that CaMKII holoenzymes are required for binding to GluN2B, suggesting that this interaction may require multiple subunits. Still, it is unknown what is the required stoichiometry and subunit geometry required for CaMKII-GluN2B binding. Initial results suggest that the holoenzyme rules for pT286 and GluN2B binding are fundamentally different. Therefore, this proposal will investigate my hypothesis that LTP versus LTD mechanisms are regulated by structurally distinct features within CaMKII holoenzymes. The approach will utilize several CaMKII “structural mutants” that have disrupted holoenzyme structure (hexamers, dimers, and monomers). These mutants will be used as tools to define the spatiotemporal dynamics of autophosphorylation within holoenzymes and the subunit stoichiometry and geometry required for GluN2B binding. The results of this proposal will provide insight into how molecular signal computation underlying the LTP versus LTD decision is encoded within CaMKII holoenzymes.

NIH Research Projects · FY 2025 · 2022-08

SUMMARY Cellular responses to external stimuli require dynamic gene regulation to facilitate rapid activation and effective resolution. Although transcriptional responses are a textbook mechanism for stimulus-induced responses, post- transcriptional regulation of mRNA translation and decay are the other side of the coin and are essential for rapid and adaptable stimulus-induced responses. For example, immune activation generates a prototypical temporal expression pattern for cytokine mRNAs known as an impulse response, which is a rapid pulse-like increase and decrease in expression. The destabilization of cytokine mRNAs is required for proper resolution of the impulse response and prevention of excess cytokine production. It is increasingly recognized that RNA-binding proteins (RBPs) play critical roles in achieving an appropriate stimulus-induced gene expression response by regulating RNA processing, decay, and translation of target RNAs. The over-arching goal of our research program is to understand how RBPs temporally coordinate stimulus-induced gene expression and cellular phenotypes. The model system we will use to study how dynamic RBP-RNA regulatory interactions temporally coordinate stimulus-induced gene expression is human adrenal steroidogenesis. This is a classic ligand-induced system in which the small peptide Angiotensin II (AngII) binds to its receptor in adrenal zona glomerulosa cells to stimulate the production of aldosterone, the master regulator of blood pressure. Thus, tight control of regulatory timing is required since it must be rapidly produced de novo from cholesterol in response to AngII. We have recently demonstrated that AngII treatment of immortalized and primary adrenocortical cells results in an impulse response. Furthermore, we found specific RBPs and regulated RNA decay facilitates the rapid implementation and resolution of the AngII impulse response. In this proposal, we will test the hypothesis that MSI2 and ZFP36L2 are counteracting forces that, respectively, potentiate and attenuate the kinetics of stimulus-induced mRNA levels by modulating target mRNA decay and translation, to ensure the proper timing and amplitude of the impulse response. We will utilize cutting edge methods to gain temporal and quantitative insights into how stimulus-induced changes in RBP binding determine changes in target mRNA expression responses and ultimately cellular phenotype. The use of innovative approaches we use will help shift the field from a static to a dynamic view of RBP-mRNA interactions and achieve mechanistic insight on how regulatory RBP-mRNA interactions control mRNA fate decisions to govern cellular responses.

NIH Research Projects · FY 2025 · 2022-08

Project Summary The project’s overall goal is to determine important biological characteristics and investigate therapeutic options for pediatric high-grade gliomas (PHGG), the most aggressive of childhood central nervous system tumors and most common cause of childhood cancer mortality. PHGG survival rates are less than 5% for the subtype diffuse midline glioma and 20% for hemispheric histone 3-wild type (H3-wt) PHGG. PHGG are highly invasive and often grow diffusely among normal cells, limiting surgery as a therapeutic option. Radiation therapy (RT) is transiently effective, but the tumors nearly always recur. Despite hundreds of clinical trials, no chemotherapy has shown a definitive survival benefit in PHGG. Effective PHGG therapies are critically needed. PHGG likely originates from stemlike tumor initiating cells (PICs). PHGG tumors comprise several distinct cell types of glial origin, in varying proportions. This tumor heterogeneity complicates understanding PHGG tumor biology and designing therapies. Aim 1 will investigate how each distinct cell type in PHGG contributes to overall tumorigenesis in a mouse model. Single-cell RNA-Seq (scRNA-Seq) analysis of orthotopic patient derived PHGG xenografts (PDX) will be used to define the cell types present and identify differentially regulated oncogenic pathways that drive their growth. Pathway expression will be knocked down by targeting key effector genes with shRNA using stable lentiviral transduction. The effect on tumor growth will be evaluated using survival, histology and single-cell RNA-Seq. Aim 2 will perform lineage tracing to determine whether a single PIC cell type produces all of the proliferating cell types that comprise PHGG. Lineage tracing will be performed in a mouse PDX model. Singlecell genomic DNA sequencing will be performed on PDX tumors. Mutational signatures consisting of single and multiple nucleotide variations as well as copy number variation will be used to define each cell type. Conserved patterns of mutation among cell types will be used to determine the hierarchical relationships among cell types. Once the lineage relationships are worked out, resistance to RT will be studied in the PDX model. RT is the most consistently effective therapy against PHGG but works only temporarily before cells regrow. RT resistance by cell type will be determined based upon differential survival of cell types versus control following RT. Drug screening of resistant cell types to identify radiation sensitizers will be performed. The candidate drugs will be combined with RT to investigate their effectiveness at increasing the duration of the RT effect.

NIH Research Projects · FY 2025 · 2022-08

Project summary Steroid receptors are a subset of nuclear receptor (NR) transcription factors that are found only in vertebrates and regulate essential functions such as organismal development and reproduction, while also impacting aging, tumorigenesis, and cancer progression. Small lipophilic hormones bind to steroid receptors for estrogens (ER- alpha and ER-beta), progesterone (PR), glucocorticoids (GR), and androgens (AR) and transmit their signal through gene regulation. Since these small molecules can be synthetically modified and manufactured, a variety of pharmaceutical drugs provide critical medicines for diseases including metabolic disorders, reproduction, and cancer treatment. NR mechanism of action has been mostly studied at RNA polymerase II (Pol II) transcribed genes including protein-coding mRNAs and small/long non-coding RNAs. Multiple NRs act in complexes on DNA to activate or repress Pol II transcription. However, NR controlled cell transcriptomes are often not always tightly correlated with the proteome due to post-transcriptional regulation that is not completely understood. We have uncovered a second layer of coordinated NR activity through regulation of RNA Polymerase III (Pol III) transcribed genes. Pol III transcribes small RNAs essential for translation of mRNAs into protein including tRNAs and 5S rRNA and is a major node for controlling cell growth, stem cells, aging, and cancer. Negative regulation of Pol III is commonly through the conserved repressor Maf1. Very little is known concerning how NRs regulate Pol III in more complex mammalian cells and organisms. We discovered using genome-wide analyses of PR chromatin binding in ER+PR+ breast cancer cell lines and tumors that PR localizes at multiple tRNA genes. PR associates with the Pol III complex and decreases tRNA levels and protein synthesis. Progesterone recruits PR and retinoic acid receptor alpha (RARα) to tRNA genes near a conserved DNA sequence resembling an NR binding half site. Our hypothesis is that PR regulates Pol III transcription of tRNA genes through crosstalk with RARα and recruitment of Maf1 resulting in decreased levels of target tRNA genes and selective translation. Aim 1 will determine how PR associates at tRNA genes, the role of RARα and other steroid receptors, and the NR binding half site. Aim 2 will determine the role of Maf1 in PR modulation of Pol III transcription. Aim 3 will define hormone-induced changes in the tRNA pool and the impact on translational efficiency. Regulation of Pol III is vastly understudied compared to Pol II and crucial for normal and oncogenic cell phenotypes. Results of this study will define a novel mechanism of NR action at Pol III genes that will help explain i) an additional layer of hormone regulation that contributes to selective mRNA translation, and ii) how NRs converge on multiple cell polymerases to impact cell growth, differentiation, stemness, and tumor progression.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Cardiovascular disease (CVD) and diabetic kidney disease (DKD) are the leading causes of morbidity and premature death in youth and adults with type 1 diabetes (T1D). Recent advances in continuous glucose monitoring (CGM) and automated insulin delivery systems have facilitated improved glycemic control, but the residual risk of CVD and DKD remains high. Obesity and insulin resistance (IR) have also accompanied intensive glycemic therapy and may accentuate arterial stiffness and endothelial dysfunction, each of which is known to predict CVD in T1D. Thus, studies are needed to explore the cardio-renal impact of new adjunctive therapies in T1D informed by the transformative cardiovascular outcome trials in type 2 diabetes (T2D). Glucagon-like peptide-1 receptor agonists (GLP-1RAs) mitigate major adverse cardiac events in adults with T2D and support weight loss. Our group has characterized subclinical cardiorenal disease in young persons with T1D, reporting abnormalities in cardiac function, central arterial stiffness, endothelial function, kidney function, and insulin sensitivity. These cardiorenal abnormalities were also worse with increasing BMI. Our group has also documented attenuated subclinical cardiac dysfunction and aortic stiffness with GLP-1RA without increased risk of hypoglycemia or diabetic ketoacidosis, in both adults with T2D and in animal models of diabetes. To date, limited data exist regarding CVD, IR or DKD-related outcomes in young adults with T1D in response to GLP-1RA. Indeed in T1D, studies with GLP-1RA have focused primarily on weight and glucose lowering. Thus, there is a gap in our understanding of the cardiorenal impact of these agents in T1D. To evaluate the effects and underlying mechanisms of GLP-1RA on cardiovascular and kidney function as well as insulin sensitivity in T1D, we propose a 6-month randomized, placebo-controlled, double-blind study in 52 young adults with T1D (ages 18-40 years) using once weekly subcutaneous semaglutide as a mechanistic probe. The primary outcomes will be change in central and peripheral pulse wave velocity (PWV) by aortic MRI and SphygmoCor. Additional outcomes will include subclinical cardiac function by cardiac MRI, endothelial function by flow mediated vasodilatation (FMDBA), insulin sensitivity by hyperinsulinemic euglycemic clamps, intraglomerular hemodynamic function by iohexol and p-aminohippurate clearance, albuminuria by urine albumin-to-creatinine ratio, and glycemic variability by CGM. Innovative translational assessments of nitric oxide (NO) bioavailability, endothelial NO synthase (eNOS) activation, reactive oxygen species (ROS)/oxidative stress from endovascular J-wire biopsies and endothelial glycocalyx from sublingual assessments will provide mechanistic insight.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT Background: Sexual dysfunction (SD) is estimated to occur in up to 50% of childhood cancer survivors (CCS) and has important implications for quality of life and mental health. Effective management of SD in AYA-CCS is substantially limited due to lack of screening and detection of SD in cancer survivorship settings. Adolescent and young adult-aged CCS (AYA-CCS) may be particularly vulnerable to SD and its under-recognition due to developmental challenges. This study addresses this critical barrier and serves as the first step in progressing toward the long-term goal of improving health-related quality of life in AYA CCS through improved screening, recognition and treatment of SD among CCS. Candidate: The proposed award will enable the candidate, Dr. Sopfe, to develop the skills necessary to progress toward her career as an independent researcher leading high quality pragmatic trials. Training: Specifically, Dr. Sopfe will gain experience in Dissemination & Implementation (D&I) science, pragmatic clinical trials, and mixed methods, which will propel her toward a research career aimed at developing and disseminating interventions to improve quality of life for CCS. In addition to experiential training through the research plan, Dr. Sopfe will benefit from a multidisciplinary mentorship team: Primary Mentor, Dr. Peterson (hybrid trials, qualitative research, patient-reported outcomes); Co-Mentor, Dr. Chow (pediatric cancer survivorship, clinical trials); and Methods Mentor, Dr. Studts (D&I, mixed methods). Dr. Sopfe’s training plan includes formal didactic coursework, workshops, and career development programs. Research Plan: This proposal’s overall objective is to use patient and provider stakeholder input to refine and test an approach to SD screening for AYA CCS in a clinic setting. Informed by D&I frameworks, this research proposal will used mixed methods and iterative testing and modification to refine an SD screening approach (Aim 1). This approach will be then tested in a pilot type 1 hybrid effectiveness-implementation study (Aims 2 and 3), employing mixed methods to assess patient and provider outcomes. Environment: The environment for this project is exceptional with a strong academic Section of Pediatric Hematology/Oncology/BMT at the University of Colorado, the resources of the University of Colorado Adult and Child Consortium for Health Outcomes Research and Delivery Science (ACCORDS) (D&I Science Program, Qualitative & Mixed Methods Research Core). The clinical trial will occur through the 1) Children’s Hospital Colorado (CHCO) HOPE Survivorship Program; CHCO Neuro-Oncology group; and 3) Seattle Children’s Hospital Cancer Survivorship Program. This multiclinic design will ensure successful recruitment as well as increase generalizability of findings. Impact: The proposed study will serve as a necessary first step in improving screening, detection, and treatment of SD in AYA-CCS; future work will include establishing broader effectiveness our screening approach through a multicenter randomized trial, as well as development, testing, and dissemination of multidisciplinary interventions for SD in this population.

NIH Research Projects · FY 2025 · 2022-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. This application describes a new Training Program at the University of Colorado (CU) entitled “Interdisciplinary Training in Musculoskeletal Research”. The overarching mission of this new Program is to provide comprehensive interdisciplinary training in musculoskeletal science with the aim of developing the next generation of outstanding musculoskeletal investigators. The Program will be administratively anchored in the Colorado Program for Musculoskeletal Research (CPMR) and the Department of Orthopedics on the Anschutz Medical Campus. It includes 32 senior and junior preceptors from 18 basic science and clinical departments across all 4 CU campuses - Anschutz, Boulder, Denver, and Colorado Springs. Preceptors and their labs are coordinated in the Program to provide research, didactic education and mentoring to 4 predoctoral and 2 postdoctoral trainees in conjunction with a broader cross section of unseated trainees pursuing advanced education within the CPMR. To accomplish this mission, we have assembled a Program that evaluates and recruits top talent from trainee pools and provides a series of formalized education components that are conjoined with top-notch research training with world class preceptors. Beyond laboratory research, formal programming includes a five-lecture series Musculoskeletal Science Curriculum, the Mack Clayton Seminars (preeminent visiting scientists), Work-in-Progress Meetings and Journal Clubs, the Annual CPMR Symposium and D’Ambrosia Lectureship, a weekly Specific Aims Development Meeting, and preferential and subsidized access to Research Core Services for trainees and their preceptors. University-level education opportunities constructed and supported by the Colorado Clinical and Translational Sciences Institute are layered in, including training in mentoring, biostatistics, and research management. Program activities will be developed, implemented, executed, and supervised by a hierarchy of oversight Committees that collaborate with Program leadership to administer all aspects of activity. These Committees include Trainee Selection and Progress, Curriculum, Clayton Seminars, Annual Symposium and Trainee Feedback. The Program Directors will coordinate in real time with these Committees for overall Program management, collectively reporting to an Internal and an External advisory board and an Executive Committee to facilitate overall Program oversight, evaluation, problem solving, and vision setting. Overall, the training enabled through this new Program, which includes a focus on mentoring trainees towards independent funding, will be a springboard to development of the next generation of collaborative and innovative musculoskeletal investigators aiming to translate basic discoveries into novel human therapies.

NIH Research Projects · FY 2026 · 2022-07

PROJECT SUMMARY Our proposal to establish the Colorado Resource Center for Tribal Epidemiology Centers capitalizes on the Centers for American Indian and Alaska Native Health’s 30+ year, nationwide experience providing high quality, scientifically grounded, culturally informed training and technical assistance to hundreds of Native programs through resource centers that addressed needs similar to those highlighted in RFA-MD-21-003. We marry this programmatic focus with an equally superior record of preparing Native investigators to compete for NIH- sponsored research and, thereby, enhance the nature, extent, and quality of the science undertaken by Tribal Epidemiology Centers (TECs). Accordingly, the specific aims of our Resource Center-TEC are to: 1) Jointly plan, issue, monitor, and evaluate subawards of varying degrees of support to each TEC consistent with local priorities and needs that progressively expand their capacity to engage in the full spectrum of research activities ranging from data acquisition, management, and analysis to data visualization, interpretation, and reporting. 2) Provide technical assistance to each TEC to design, implement, and analyze both primary data acquisition as well as secondary data analytic studies consistent with local priorities, and to prepare as well as disseminate lay, program, policy, and scientific reports of findings to a wide array of audiences. 3) Enhance the research capacity of TEC personnel across all levels of preparation by drawing upon an extensive body of existing instructional materials and experience to offer: a. bimonthly interactive, 2-hour didactic sessions on timely data science topics; b. on-line, short-course offerings that target foundational and intermediary knowledge and skills regarding data collection, linkage, management, and analysis; c. once annually, a 6-month, mentored grant-writing course for senior TEC personnel—tailored to the interests and priorities of their respective key stakeholders—to increase the number of submitted and funded NIH R-series grant applications, and d. a year-long research career development program for early-stage investigators to add to and retain them in the health disparities scientific workforce. 4) Promote collaborative research between the TECs and external investigators by disseminating information about sponsored research opportunities and related resources and building upon our current Satellite Center research network led by senior, NIH-funded American Indian and Alaska Native health disparities scientists at 7 adjacent major institutions, and other organizational partnerships. Recent calls to action by advocates underscore the important contribution of research to address the health priorities of Native peoples. This Resource Center will enable TECs to bring this vision to fruition.

NIH Research Projects · FY 2025 · 2022-07

Project Summary/Abstract Candidate Goals and Mission Relevance: The applicant’s broad, long-term objective is to investigate how high- (circuit/behavioral) and low- (subcellular/molecular) level organizational principles of the brain cooperate to drive learning. The proposed research activities will build a foundation for this long-term goal and, in so doing, will promote BRAIN 2025 Report goals by integrating new technological and conceptual approaches to causally link intracellular Ca2+ release (ICR) from endoplasmic reticulum (ER) to neural activity dynamics and behavior. Project description: Dendritic Ca2+ is central to neural plasticity mechanisms allowing animals to adapt to the environment. ICR has long been thought to shape these mechanisms. The applicant recently carried out the first investigation of ICR in mammalian neurons in vivo to uncover how this subcellular phenomenon shapes experience-dependent feature selectivity across the dendritic arbor of pyramidal neurons (PNs) in mouse hippocampal area CA1. This work raises important questions regarding when, where, and how ICR is engaged to support learning. The applicant will address these questions in the following Aims: Aim 1. Characterize plasticity-associated ER Ca2+ dynamics in dendrites in vivo (K99): To achieve this Aim, the applicant will perform simultaneous dual-color, dual-plane in vivo 2-photon imaging of cytosolic and ER- resident Ca2+ in dendrites of single CA1 PNs during head-fixed spatial navigation of novel virtual environments. Aim 2. Define the synaptic logic tying intracellular Ca2+ release to in vivo synaptic plasticity (K99/R00): The applicant will first create a novel molecular tool to optogenetically induce ICR (Aim 2.1; K99). The applicant will then combine this precise interventional tool with single-cell imaging, inducible blockade of presynaptic release, and optogenetic dampening of ICR to dissect the synaptic logic by which ICR participates in plasticity induction in behaving mice. (Aim 2.2; R00). Aim 3. Dissect excitatory circuit-molecular mechanisms driving intracellular Ca2+ release in vivo (R00): The candidate will optogenetically activate specific excitatory projections onto distinct dendritic compartments of single CA1PNs while monitoring ER Ca2+ dynamics in behaving mice. Local pharmacological manipulations will dissect contributions of the two canonical pathways that convert presynaptic excitatory input to postsynaptic ICR. Career development plan: The applicant will extend a highly complementary Co-Mentorship arrangement between Drs. Franck Polleux and Attila Losonczy who possess deep expertise in cellular/molecular/genetic and in vivo/behavioral approaches, respectively. The applicant will receive robust consultative support from Dr. Stefano Fusi of Columbia’s Center for Theoretical Neuroscience and Dr. Darcy Peterka, Director of Cellular Imaging at Columbia’s Zuckerman Institute. The applicant’s research and transition to independence will benefit from this strong mentorship team, state-of-the-art facilities, all necessary equipment, and numerous Professional Development resources offered through the Columbia Office of Postdoctoral Affairs, the Zuckerman Institute, and the BRAIN Initiative.

NIH Research Projects · FY 2025 · 2022-07

Summary Growing evidence points towards the contribution of altered brain microcirculation to cognitive impairment and dementia observed in Alzheimer’s disease (AD) and AD-related dementia (ADRD). Yet, the lack of approaches to image the small cerebrovasculature and investigate its function has hampered our progress in understanding the pathological sequence of vascular cognitive impairment and dementia (VCID). The earliest signs of AD and VCID in patients and mouse models typically involve deficits in spatial and short-term memory—cognitive functions that are critically sustained by synaptic plasticity in the hippocampus. Neurons have limited energy reserves and thus rely on a “just-in-time” neurovascular coupling (NVC) strategy in which active regions signal to the microvasculature to locally dilate and increase local blood flow. Patients and mouse models of AD or CADASIL, a monogenic archetypal form of VCID, show an early deterioration in NVC. Our previous studies have identified a molecular defect at play in capillary endothelial cells and developed a therapeutic approach that acutely restores NVC in the mouse model of AD and CADASIL. Specifically, we found that systemic injection of phospholipid PIP2 is sufficient to rescue neurovascular deficits by enabling Kir2.1 channels to act as sensors of increases in external K+—a product of neuronal activity—and transduce this into a vasodilator electrical signal that rapidly propagates to upstream arterioles, driving vasodilation to produce local hyperemia. Our multidisciplinary team, with complementary expertise in cutting-edge imaging of brain microcirculation and synaptic plasticity underlying learning and memory processes, will test the hypothesis that NVC restoration will mitigate the synaptic plasticity deterioration in the hippocampus, and its behavioral consequences, observed in AD. We further propose to investigate and compare these functions in CADASIL, a vascular driven form of ADRD. To attain this goal, we will advance our PIP2-based strategy to chronically restore NVC in AD and CADASIL models, and assess the treatment efficiency by developing innovative imaging approaches ex vivo, with a novel intact capillary-arteriolar (CaPA) preparation established by our group, and in vivo using implanted graded-index (GRIN) lenses combined with 2-photon microscopy to investigate NVC in the hippocampus. Ultimately, we will measure the effect of NVC rescue on hippocampal synaptic plasticity deterioration caused by AD and CADASIL conditions, and use contextual fear conditioning as a behavioral readout. Completing this study will help elucidate the mechanisms linking NVC dysfunction to dementia in AD/ADRDs, and NVC restoration as a potential therapy. The proposed work has the potential to provide a paradigm-shifting view on how brain microcirculation sustains learning and memory processes.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT There has been more than a 30% increase in opioid overdose deaths in the last year. Bystander access to naloxone medication effectively reverses opioid overdose deaths with limited adverse events. However, current naloxone distribution strategies are missing individuals at high risk of opioid overdose. Vending machines are an innovative strategy shown to increase reach of harm reduction services in Europe. Young adults who witnessed or experienced an opioid overdose expressed a desire for naloxone vending machines but identified features of the built environment (including safety and location) as perceived factors contributing to use. There is widespread interest in naloxone vending machines in the U.S. Yet, implementation challenges have slowed down or limited adoption including navigating stakeholder approvals and identifying appropriate locations for placement. The aim of this proposal is to study a community-initiated, stakeholder engaged adaptation of naloxone distribution, VEnding machine Naloxone Distribution for Your community (VENDY), to increase the reach of naloxone in underserved populations at-risk of opioid overdose. Study Aims include: Aim 1: To refine the VENDY program in 3 underserved communities (2 urban and 1 rural) using an iterative user centered design (UCD) implementation strategy and stakeholder engagement to increase reach, implementation, and sustainability. We will use iterative usability and message testing to refine the program with community members who misuse opioids. We will conduct surveys and qualitative interviews with stakeholder to refine the UCD implementation strategy for future use. Aim 2: Conduct a 6 month pilot assessment of the VENDY program in 2 urban and 1 rural community. The pilot includes a midcourse adaptation and evaluation of reach, effectiveness, adoption, implementation and maintenance (RE-AIM) outcomes. Aim 3: Identify factors in the social and built environment contributing to reach and implementation of the VENDY program using photovoice qualitative evaluations with organization implementers and community members currently using opioids. Photovoice evaluations will occur in potential locations in Aim 1 to inform VENDY placement and in the VENDY location in Aim 2 to inform stakeholder adaptations. Community-based approaches such as VENDY are particularly important to reach opioid users not currently engaged with health systems. To accomplish these aims, Dr. Wagner will pursue training in 1) user centered design, an innovative implementation strategy, 2) conduct of pragmatic trials in diverse settings, and 3) assessment of the built and social environment and its effect on reach and implementation. This mentored research project and career development plan are designed to prepare Dr. Wagner to become a leader in the adaptation of effective interventions addressing substance use to better reach underserved populations and to produce sustainable implementation models.

NIH Research Projects · FY 2024 · 2022-07

PROJECT SUMMARY/ ABSTRACT Despite extensive efforts aimed toward the development of improved molecular therapies targeting acute myeloid leukemia (AML), clinical outcomes remain poor. Of particular interest, is the necessary and selective therapeutic targeting of disease initiating leukemia stem cells (LSC). The Jordan laboratory has reported that LSC are functionally reliant upon BCL2 for cellular oxidative phosphorylation (OXPHOS) requirements. Targeting BCL2 with venetoclax (Ven) in combination with azacitidine (Aza) has clinically delivered significant responses in newly diagnosed AML patients, however both upfront refractory and relapsed diseases are still a major obstacle. Notably, we show that Ven/Aza resistant AML express elevated MCL1 protein and OXPHOS levels. Moreover, the Jordan laboratory have recently reported that pharmacologic perturbation of MCL1 in resistant specimens leads to a selective decrease in OXPHOS output as well as reduced LSC functional ability as measured by engraftment of immune deficient mice. Continued analysis of Ven/Aza resistant AML highlighted a significant increase in mitochondrial fission promoting DRP1 phosphorylation as well as in metabolomic enrichment of fatty acid oxidation. Thus, we hypothesize that MCL1 specifically drives Ven/Aza resistance by promoting mitochondrial fission and β-oxidation. As this proposal aims to define the mechanisms through which MCL1 uniquely influences therapy resistance in AML, our studies will largely utilize Ven/Aza resistant primary AML specimens to interrogate the specific role of MCL1 in regulating mitochondrial function through fission and β-oxidation. Successful completion of these studies will generate a detailed and mechanistic understanding of the non-canonical roles for MCL1 in regulating mitochondrial morphology and lipid metabolism, while also providing alternative approaches for therapeutic intervention in therapy resistant AML.

NIH Research Projects · FY 2025 · 2022-07

ABSTRACT Human leukocyte antigens (HLA) and killer cell immunoglobulin-like receptors (KIR) are critical facets of the human immune system. Interactions of KIR, expressed by natural killer cells (NK cells), with HLA class I, expressed by most tissue cells, modulate immune cell functions. Variations in the highly polymorphic KIR and HLA genes are linked directly to NK cell functions and have profound impact on human health, including associations with autoimmunity and neurological disease, severity of infectious disease, pregnancy syndromes, and risk of cancer. Our ability to resolve the mechanisms of immune-mediated disease, to develop personalized medicines, including immunotherapies, and to match organ donors with recipients relies on our ability to accurately characterize KIR and HLA class I diversity. Despite this crucial importance, there is a lack of knowledge concerning the extent and nature of KIR and HLA class I diversity worldwide. This deficit includes the Eastern Hemisphere, which encompasses half the world's population, and multiple underrepresented groups in the USA. During this project we will fill these gaps in this knowledge through determining the characteristics and functional consequences of KIR and HLA class I diversity across the entire Eastern Hemisphere. We will examine 15,612 individuals representing 51 discrete populations, including indigenous populations from East Asia, South Asia, multiple Pacific Islands and Oceania. To overcome difficulties in analyzing these complex genomic regions, we developed a targeted sequencing and bioinformatics approach to analyze KIR and HLA class I genes at high throughput and resolution. To analyze an additional 22,905 individuals, we will develop an imputation algorithm to determine high-resolution KIR alleles from whole-genome SNP data. We will construct imputation panels specifically for these populations who have been neglected in previous analyses. This goal will be made possible through the extensive training data generated. We will then characterize the distribution of KIR and HLA haplotypes across the Eastern Hemisphere. We will examine how the geographic patterns of KIR and HLA diversity have been shaped by natural selection and investigate the impact of adaptive introgression and admixture specifically focused to the KIR and HLA loci in our study populations. We will determine the functional properties of those variants we have identified as targeted by natural selection. We will pursue these aims implementing innovative laboratory and analytical tools. These developments include CRISPR/cas9 targeting of long-read sequencing to examine KIR structural diversity, and methods both to identify alleles subject to natural selection, and the mode of selection acting on them. The expected outcome of this work is the genetic and functional characterization of HLA and KIR across multiple human populations, and a comprehensive understanding of how this variation is geographically distributed and shaped by natural selection. This work will benefit investigations of immune-mediated and infectious disease and in establishment of personalized treatments for individuals both in the USA and worldwide.

NIH Research Projects · FY 2024 · 2022-07

Project Summary Craniofacial development involves complex signaling to coordinate tissue organization to form the head and face, and disruptions in this process result in common congenital malformations. A key question in this field is how external stimuli lead to gene expression changes required to form fully developed craniofacial structures. Signaling through the Platelet-derived growth factor receptor alpha (PDGFRα) plays a critical role in this process, as mutations in PDGFRΑ are associated with cleft lip/palate in humans. Relatedly, Pdgfra mutant mouse models develop a range of phenotypes from cleft palate to complete facial clefting. Phosphatidylinositol 3-kinase (PI3K) is the primary effector of PDGFRα signaling during skeletal development in the mouse, leading to the activation of the kinase Akt. A previous phosphoproteomic screen demonstrated that Akt phosphorylates the RNA-binding protein (RBP) Serine/arginine-rich splicing factor 3 (Srsf3) downstream of PI3K-mediated PDGFRα signaling in mouse embryonic palatal mesenchyme (MEPM) cells, leading to translocation of phosphorylated Srsf3 into the nucleus. Srsf3 is ubiquitously expressed with enhanced expression in the neural crest-derived mesenchyme and overlying ectoderm of mouse facial processes at mid-gestation. Additionally, ablation of Srsf3 in the murine neural crest cell lineage (cKO) results in a severe midline facial clefting phenotype due to defects in proliferation and survival of cranial neural crest cells. Further, RNA-sequencing of Srsf3 cKO facial process mesenchyme identified alternative RNA splicing events that were enriched for transcripts encoding protein serine/threonine kinases, suggesting that alternative splicing may serve as a novel feedback mechanism for intracellular kinase signaling. The goal of this proposal is to test the hypothesis that PI3K/Akt-mediated PDGFRα signaling regulates Srsf3 protein and RNA interactions to affect the alternative RNA splicing of transcripts necessary for craniofacial development. First, Srsf3 will be immunoprecipitated from MEPM cells in the absence or presence of PDGF-AA ligand and analyzed by mass spectrometry to comprehensively map phosphorylation changes in response to PDGFRα signaling. Further, craniofacial phenotypes will be analyzed in a Srsf3 phosphomutant knock-in mouse model to determine the role of Akt- mediated phosphorylation of Srsf3 in craniofacial development. Next, BioID2 proximity labeling and mass spectrometry will be used to identify Srsf3 protein interacting partners in response to PDGFRα signaling in MEPM cells. Finally, Srsf3-RNA interactions will be purified and sequenced in response to PDGFRα signaling in MEPM cells through enhanced crosslinking and immunoprecipitation analysis to identify direct targets of Srsf3 and determine if RNA binding and/or sequence specificity changes upon Srsf3 phosphorylation. This project will determine the molecular mechanisms by which Srsf3 activity is controlled in response to PDGFRa signaling in the facial mesenchyme, thus providing considerable insight into mechanisms underlying gene expression regulation during mammalian craniofacial development.

NIH Research Projects · FY 2025 · 2022-07

Project Summary/Abstract The rapidly emerging modalities of long-acting antiretroviral therapy (ART) for treatment and prevention are based on a one-size-fits-all dosing strategy despite these being our first experiences in real-world settings, where patient management challenges are common. To address this issue, we propose a novel pharmacologic monitoring platform for long-acting ART in real-world settings. We aim to develop and validate at-home self- collections and point-of-care (POC) testing for quantitative cabotegravir/rilpivirine (CAB/RPV) concentrations to support patient management and inform optimal use of long-acting ART and PrEP, with the vision that this platform will be widely implementable and able to accommodate next in line long-acting HIV therapies in development. CAB/RPV concentrations were influential in tightly controlled trials, even with a 1% virologic breakthrough rate. Among 1,039 participants, CAB/RPV concentrations varied by more than 10-fold at a single 4-week post-injection time point. Of 13 observed HIV breakthroughs, none occurred when concentrations of both drugs were above the median, no matter what other risk factors were present –including archived resistance mutations. On the other end of the spectrum, some patients achieved CAB/RPV concentrations well-above expected levels and could comfortably extend dosing beyond Q8W, but this possibility was not adequately investigated. In practice, CAB/RPV concentrations will be more variable than in tightly controlled trials because real-world patients miss appointments, change providers, and leave/reenter care. Additionally, within-person variability from injection-to-injection is unknown. To address these gaps and needs, we propose the following specific aims: Aim 1. Validate at-home self-collections and POC testing. Building on our preliminary data using at-home self-collections and POC testing via miniature mass spectrometry, we will develop and optimize suitable assays using samples from 30 persons with HIV (PWH) receiving Q4W or Q8W long-acting injectable CAB/RPV. Assays will be validated using FDA bioanalytical guidance. Aim 2. Classify CAB/RPV concentrations in a real-world longitudinal cohort. Through longitudinal blood collections in 50 PWH receiving Q4W or Q8W long-acting injectable CAB/RPV, we plan to classify drug concentrations into low, expected, and high categories and define specific pharmacokinetic (PK) parameters, including within- and between-person variability. Aim 3. Establish pharmacokinetic strategies for personalized dose intervals. We will identify the ideal drug concentration testing strategy to determine patient-specific PK, and in turn, the optimal dose frequency to achieve goal concentrations. We envision an easy-to-use interface where clinicians will input drug concentrations to calculate personalized dosing. This application is timely and provides innovations to keep pace with the rapidly emerging long-acting era.

NIH Research Projects · FY 2025 · 2022-07

Project Summary: Overall BRIDGE Center The promise of using Big Data for precision medicine, drug discovery, and a host of other challenges has remained elusive. Social and technical barriers have limited our ability to leverage our collective data assets to address biomedical and behavioral health challenges; Artificial Intelligence (AI) solutions present an opportunity to overcome these barriers. The NIH has established the Bridge to Artificial Intelligence (Bridge2AI, or B2AI) program to catalyze AI solutions to a set of community-defined “Grand Challenges.” This concerted effort will lay the groundwork to promote the widespread adoption of AI and to ensure that it leads to trustworthy, inclusive research innovations that have a significant, positive impact on human health. The BRIDGE Center proposal includes Administrative, Teaming, Skills and Workforce Development, and Standards Cores, chosen to coincide with the Core teams’ unique and complementary expertise in large-scale team science, standards development and dissemination, innovative training approaches, and community building. The vision for the BRIDGE Center is to engage all participants within and beyond the B2AI community through carefully considered social and technical mechanisms and operational excellence, with the goal of creating a dynamic, productive, and inclusive community that builds upon each other’s work in a deeply collaborative manner. The mission of the BRIDGE Center is to complement the Bridge2AI Data Generation Projects (DGPs) by supporting the integration, dissemination, and evaluation of Bridge2AI work products and teams. We propose to achieve that mission through three aims. Aim 1 focuses on integrating across the Bridge2AI Program. Specifically, we will foster group identity to create opportunities and incentives for transdisciplinary learning; establish human & machine-understandable standards, practices, and vocabularies across B2AI; and deploy technology to promote transparency and scalability across the B2AI program. Aim 2 creates and promotes opportunities to evaluate and improve B2AI products and activities including (1) employing community-based evaluation within and across DGPs, (2) deploying data-driven evaluation and improvement of B2AI products and processes, and (3) establishing and maintaining diversity in B2AI data, people, and team structures through a continuous process of evaluation and refinement. Aim 3 focuses on sustainable dissemination of products, knowledge, best practices and “lessons learned” from B2AI thereby ensuring broad, long-lasting distribution and impact. Taken together, these Aims will create a BRIDGE Center that fosters broad community engagement, inclusivity, and trust to successfully integrate activities and knowledge across the B2AI Program; disseminate products, best practices, and skill development materials/activities; and continually assess and improve all aspects of the Bridge2AI program with input from external stakeholder communities.

NIH Research Projects · FY 2024 · 2022-07

PROJECT SUMMARY / ABSTRACT Diabetes is a global epidemic that is expected to become even more severe. Although there are treatment options, such as exogenous insulin, one of the most promising long-term treatments is transplanting donor islets into diabetic patients. While this treatment can allow patients to maintain normal blood glucose levels for extended periods of time, there is an extreme lack in the amount of viable donor islets for transplantation. To combat the lack of suitable donor islets, researchers have begun to differentiate human pluripotent stem cells (hPSCs) into β-like cells that respond to glucose by secreting insulin, with the hopes of creating a limitless supply of transplantable pancreatic β cells. This approach has been somewhat successful; however, the current differentiation protocols lack the ability to make pure populations of β cells and often result in immature β cells and polyhormonal cell populations. More complete knowledge of the transcriptional networks and associated cofactors required in the differentiation and function of pancreatic β cells is needed to supplement the current hPSC differentiation protocols and provide the information required for better diabetes treatment options. One such cofactor is the chromodomain helicase DNA-binding protein 4 (CHD4). CHD4 is the motor protein of the nucleosome remodeling and deacetylase (NuRD) complex and, along with histone deacetylase 1 and 2 (HDAC1 and 2), CHD4 creates condensed chromatin states, thereby repressing genes. Preliminary data has shown that CHD4 interacts with essential transcription factors in pancreatic β cells. Furthermore, my preliminary data shows that the loss of CHD4 in the β cells of mice causes hyperglycemia along with glucose intolerance. This suggests that CHD4 is a necessary transcriptional cofactor in the function of β cells. A better understanding of the role CHD4 plays in the development and function of pancreatic β cells could provide additional information to facilitate the directed differentiation of β cells from hPSCs in vitro. In this proposal, using both in vivo and in vitro experiments, I will determine the role and mechanism of CHD4 in the development and function of pancreatic β cells.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT As a vascular and interventional radiologist and a health services researcher, my long-term goal is to achieve transformative improvements in vascular procedural care through national quality of care assessments and implementing pragmatic inventions to improve outcomes. This research and training proposal will allow me to gain critical skills needed to establish independence and launch a portfolio of scientific contributions focused on systems-level quality improvement in venous thromboembolism (VTE) related care. The goal of this research proposal is to increase timely retrieval of inferior vena cava filters (IVCF) and improve patient outcomes in the United States. Inferior vena cava filters are very commonly placed in patients with blood clots in the leg (deep venous thrombosis; DVT) or lungs (pulmonary embolism; PE), to prevent propagation to the heart, which can be fatal. Unfortunately, national retrieval rates remain low and prolonged IVCF implantation has resulted in considerable, avoidable morbidity. Multi-society guidelines recently emphasized the need to improve timely IVCF retrieval rates through institution of structured follow up programs and to better understand which program components are effective in real world settings. What is needed next is identification and testing of pragmatic strategies to improve timely IVCF retrieval in real-world settings. We propose to do this via three complimentary aims: 1) quantification of facility-level variation in IVCF retrieval across the United States; 2) qualitative characterization of high-performing institutions to identify best practices and characterize facilitators and barriers to their implementation; 3) design, pilot testing and iterative adaptation of a pragmatic intervention package to improve IVCF retrieval, informed by the existing literature and adapted with findings from aim 2. The pragmatic intervention will then be tested in multi-center hybrid implementation-effectiveness trials to follow, with the goal of improving timely IVCF retrieval and reducing complications across hospitals in the United States. My training aims closely parallel the proposed research methodology through focused education in: a) Bayesian hospital profiling, b) qualitative interviewing and analysis, and c) intervention design using an implementation science framework. This training will be accomplished through an intentional mix of structured coursework, formal workshop experience and one-on-one education with topical research experts. My mentorship team and research environment at the University of Colorado Denver (UCD) are ideally suited to this proposal. My primary mentor, P. Michael Ho MD PhD, is a national leader in cardiovascular quality assessment and pragmatic intervention design. I will additionally leverage the extensive data and methodologic core resources of Adult and Child Consortium for Outcomes Research and Delivery Science (ACCORDS) and Data2Value (D2V) initiatives at the University of Colorado to accomplish these aims.