University Of Colorado Denver

NIH Research Projects · FY 2024 · 2020-09

Project Summary/Abstract Obesity – with its many comorbid conditions – has now surpassed smoking as the leading cause of preventable death in the United States. Despite the fact that obesity is both treatable and preventable, treating the comorbidities, rather than obesity per se remains the mainstay of therapy. Importantly, obesity is being increasingly recognized not only as a risk factor for disease, but a disease unto itself. Despite this fact, <1% of people with any degree of overweight or obesity are offered anything other than lifestyle advice. Reasons for lack of weight management prioritization are extensive and complex. Effectively, the major stakeholders, including people with obesity, their clinicians and insurers, have yielded to the epidemic. Thus, there is a critical need to systematically address these diverse barriers with pragmatic approaches and evidence that facilitates the practice of weight management. To address this gap, the University of Colorado Health Family Medicine clinic in Westminster, Colorado developed PATHWEIGH. PATHWEIGH prioritizes weight management in primary care by: 1) using a designated, time-efficient flowsheet built into EPIC (the most widely utilized electronic medical record (EMR) system in the U.S.) that guides and captures history around weight gain and recommendations for weight loss, as well as practical issues related to diagnosis and billing, and 2) clinician and team training on a) use of PATHWEIGH as a tool, and b) education on current effective practices for weight management. Our pilot study demonstrated the ability to effectively implement PATHWEIGH in a primary care clinic that lead to significant improvement in patient weight loss over 18 months. The overall objective for the proposed work is to test the effectiveness of PATHWEIGH vs. standard of care in 57 primary care clinics that span diverse settings, patient populations and community contexts, as well as to examine methods to optimize implementation. Our specific aims include: 1) Evaluate the implementation of PATHWEIGH and determine its effectiveness versus control clinics using standard of care (SOC) on patient weight loss at 6 months (primary) and weight loss maintenance at 12 and 18 months (secondary) for weight- prioritized visits in primary care, 2) Identify predictors of patient weight loss and weight loss maintenance using mediator and moderator analysis, including relevant patient, provider and clinic-level variables, and 3) Examine contextual factors affecting the adoption, implementation and sustainability of PATHWEIGH using the Practical, Robust, Implementation and Sustainability Model (PRISM). To complete these aims, we will utilize a stepped wedge cluster randomized trial. Data collection and analysis methods include clinical data, surveys, observations and interviews using statistical, qualitative and mixed methods. PATHWEIGH has the potential to be a scalable, low-cost, pragmatic approach to obesity. The rationale that underlies the proposed research is that the ultimate success of PATHWEIGH relies on understanding how to implement evidence- based weight management interventions and make them most effective, used and maintained in practice.

NIH Research Projects · FY 2025 · 2020-09

PROJECT SUMMARY Even though cone beam computed tomography (CBCT) is the most commonly used volumetric image guidance modality, its role has been severely limited in the context of treatment monitoring and patient-specific treatment modifications in radiation therapy. Due to CBCT’s poor image quality, clinicians cannot clearly visualize soft tissues to assess anatomical changes, thus affecting their clinical decision-making. Moreover, tools for treatment monitoring, such as deformable registration and dose calculation, do not function robustly with today’s CBCT images due to the lack of CT number accuracy. Scattered radiation remains to be the fundamental problem in improving CBCT image quality. Thus, in this project, we propose the two-dimensional antiscatter grid (2D Grid) as a novel device to address the scatter problem and achieve high-quality CBCT images that are suitable for treatment monitoring. Our device has fundamentally different architecture and fabrication than existing antiscatter grids for CBCT. Due to its optimized grid structure, our 2D Grid provides both higher primary transmission and better scatter rejection performance than today’s state-of-the-art antiscatter grids. Due to its favorable primary transmission and scatter rejection performance, our 2D Grid improves the contrast-to-noise ratio and CT number accuracy to levels not achievable with existing antiscatter grids. We hypothesize that our 2D Grid will provide significantly better soft tissue visualization and CT number accuracy, and deformable registration algorithms are expected to perform significantly better. To test our hypotheses, we will develop and optimize data processing methods for 2D Grid implementation in CBCT (Aim 1). Subsequently, we will fabricate 2D Grid prototypes and evaluate their performance in clinical CBCT systems for photon and proton therapy (Aim 2). Following phantom based evaluations, we will conduct a prospective clinical trial to evaluate the clinical utility of improved image quality (Aim 3). We will perform observer studies to quantify the improvement in soft tissue visualization with respect to existing clinical CBCT and gold-standard Helical CT, assess the improvement in accuracy of deformable image registration algorithms, and evaluate the improvement in consistency of image intensity and texture features. While our application is focused on radiation therapy, the 2D Grid can play a key role in other CBCT applications, such as interventional radiology, extremity imaging, and intraoperative imaging. Due to its improved low-contrast visualization performance, our 2D Grid may also allow reduction of the imaging dose in CBCT.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT The long term goal of the proposal is to determine the role of pulsatility during cardiopulmonary bypass (CPB) for preserving endothelial function and preventing organ injury after cardiac surgery. More than 400,000 people undergo cardiac surgery in the United States each year which allows them to live longer and more fulfilling lives, but if the surgery is complicated by acute kidney injury (AKI) mortality increases up to 10-fold. Dr. Clendenen is a Cardiothoracic Anesthesiologist dedicated to improving clinical outcomes after cardiac procedures and this K23 proposal is designed to provide him with the structured mentorship and training necessary to become a leader in innovative clinical trials designed to improve patient outcomes. He has developed a collaborative mentorship team led by Dr. Adit Ginde who is an Emergency Medicine Physician and national PI for a large PETAL network clinical trial with 3,000 participants, with co-mentorship provided by Dr. Sarah Faubel, an expert in renal disease research, and Dr. Kristen Jablonski Nowak, a recognized leader in endothelial function. Methodological advisors include Dr. Alex Kaizer (Biostatistics) and Dr. Kerrie Moreau (Vascular Biology). This proposal leverages the collective expertise of an established mentorship team and the unique environment at the University of Colorado Anschutz Medical Center that incorporates a tertiary referral center for cardiac surgery in the setting of a large public research University. These ample resources will allow Dr. Clendenen to address key barriers to reducing morbidity and mortality after cardiac surgery with an adaptive clinical trial comparing the incidence of AKI after cardiac surgery with pulsatile versus non-pulsatile CPB. He will accomplish this by determining the effect of pulsatile CPB on post-operative endothelial function in a prospective observational cohort (Aim 1), determining the timing of endothelial dysfunction and renal injury after cardiac surgery in the same cohort (Aim 2), and determining the optimal level and efficacy of pulsatility during CPB to prevent AKI with an adaptive seamless phase IIa/IIb clinical trial (Aim 3). These specific aims require Dr. Clendenen to develop expertise in assessing clinical endothelial function and AKI risk and designing and completing an adaptive clinical trial. He will accomplish this with a focused career development plan including coursework on clinical trial design and biostatistics alongside structured training in assessing endothelial function to measure the effects of CPB. Completion of the proposed project will provide the necessary mentorship, training, and experience required to allow Dr. Clendenen to become a leader in adaptive clinical trials to improve outcomes after cardiac surgery.

NIH Research Projects · FY 2025 · 2020-09

Project Summary/Abstract Alzheimer's Disease and Related Dementias (ADRD) are chronic, disabling cognitive impairments that will affect an estimated 12 million Americans by 2050. Undetected, these changes can lead to financial losses from elder abuse and fraud, forgetting to pay bills, and compromised financial decision-making. Often, cognitive impairment is not discovered until after patients have lost significant sums of money and experienced additional functional decline. One possible solution is to recognize the early signs of ADRD in financial data. In pilot data (R21AG053698), we linked 20 years of Medicare claims to quarterly credit reports, demonstrating that ADRD patients without a spouse/partner are more likely to miss bill payments, develop subprime credit, and experience adverse financial events for years prior to their diagnosis, a pattern unique to ADRD. Our prior work suggests that banking and credit data can be used to screen for dementia in the clinical setting, and to protect patients and families from ADRD-linked financial exploitation and other losses, including health effects of dealing with additional financial stress. We propose to test these hypotheses in the following aims, using newly created financial data linkages and partnering with patients, government, and industry: Aim 1: Test whether and when unique financial symptoms of ADRD are present in credit data prior to clinical diagnosis in coupled households. Aim 2: Test whether unique financial symptoms of ADRD can be observed in banking and brokerage data before accountholders experience elder mistreatment, fraud, and diminished capacity. Aim 3: Compare 4-year rates of mortality and hospitalization among spouses of ADRD patients with and without adverse credit events by the time of diagnosis. Aim 4: Assess the feasibility and ethical implications of using financial data to detect ADRD. We will use a 20-year panel of Medicare claims linked to consumer credit reports for Aims 1 and 3 and more than 10 years of account information from a large US bank in Aim 2. We will compare ADRD to other health conditions and sources of elder mistreatment to determine whether it has a unique financial presentation. Our study team includes an interdisciplinary group of physicians, economists, ethicists, and health services researchers with a long history of collaboration in partnership with the Federal Reserve, patient and industry stakeholders. Findings from this study will provide the most comprehensive information to date on the prevalence and magnitude of financial losses and elder mistreatment prior to ADRD diagnosis, as well as their impact on spousal health. This information is critical for many public and private decisions ranging from when and whether to begin screening for ADRD, the potential role of financial institutions in protecting clients who may be unaware of their early cognitive decline, and whether consumer data are sufficiently informative about health to require additional privacy protection.

NIH Research Projects · FY 2024 · 2020-09

Abstract New methods to study heterogeneity at cellular resolution in complex tissues are transforming our understanding of human biology and disease. These approaches measure differences in gene expression, chromatin accessibility, and protein levels across thousands to millions of cells to understand developmental trajectories of tissues, tumors, and whole organisms. But these methods rely on measurements of static levels of DNA, RNA, and proteins, and fail to capture dynamic biochemical activities that underlie complex cellular functions. Instead of developing more direct readouts of cellular function, the field has focused on inferring functional status from measurements of mRNA abundance and chromatin accessibility in single cells. To accelerate the study of biochemical heterogeneity among single cells, we developed functional assays as a new modality for single-cell experiments. Instead of measuring the abundance of molecules—i.e., levels of DNA, RNA, or protein—from single cells and predicting cell functional states (e.g., cell cycle phase), our key innovation is to directly quantify enzymatic activities in single cells by measuring the conversion of substrates to products by single cell extracts in a high-throughput DNA sequencing experiment. Our approach is compatible with existing platforms that measure gene expression in thousands to millions of individual cells and enables many different enzymatic activities to be measured simultaneously.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY: Nontyphoidal Salmonella infections are frequently associated with diarrhea in healthy people. Some serovars such as Typhimurium are also common causes of bacteremia in HIV-infected people, and life- threatening disseminated complications in immunocompromised individuals with defects in neutrophils, macrophages or CD4 T cells. Sensor kinases and their cognate response regulators in two-component systems orchestrate many virulence programs in Salmonella and many other pathogenic bacteria. In the canonical activation of two-component systems, the sensor kinase is phosphorylated in response to cues encountered during colonization and infection of the mammalian host. The transfer of the phosphoryl group from the sensor kinase to the receiver domain of its cognate response regulator turns on virulence programs essential for bacterial pathogenesis. We have made the unexpected discovery that two-component response regulators are controlled by previously unknown allosteric interactions with thioredoxin. Our research has shown that thioredoxin post-translationally controls several response regulators such as OmpR, PhoP and SsrB, all of which govern key aspects of Salmonella pathogenesis. Strikingly, the post-translational control exerted by thioredoxin on two-component signaling does not rely on the universally conserved thiol-disulfide oxidoreductase enzymatic activity of this ancestral protein, but is contingent upon a hitherto uncharacterized hydrophobic interfacial surface that has been preserved throughout the evolution of thioredoxin in bacteria, archaea and eukaryotes. Our investigations indicate that most contributions of thioredoxin to Salmonella pathogenesis are independent of its oxidoreductase activity but are carried out by this newly discovered interfacial surface. The proposed research will test the hypothesis that thioredoxin leverages the binding attributes of a conserved hydrophobic patch to establish protein-protein interactions with multiple response regulators, thereby exerting broad post-translational control of two-component signaling. Specifically, we will identify the interfacial residues that mediate oxidoreductase-independent functions of thioredoxin, and will quantify the extent that the novel thioredoxin-binding face enables response regulators to activate Salmonella virulence programs. Our research will elucidate previously unappreciated elements in the regulation of two- component signaling, and will ascertain unprecedented, oxidoreductase-independent functions of thioredoxin. The knowledge gained on the novel function of thioredoxin will not only shed light on key aspects of Salmonella pathogenesis, but may ultimately broaden our understanding of a primordial function of universally-expressed thioredoxin proteins. Our work will also provide far reaching insight into the regulation of two-component systems, which represent a dominant signaling pathway in bacteria. Drugs that specifically inhibit interactions between thioredoxin and response regulators may inform the rational development of the next generation of antibiotics.

NIH Research Projects · FY 2025 · 2020-09

PROJECT SUMMARY/ABSTRACT Our re-submission proposal, “Reducing Asthma Attacks in Disadvantaged School Children with Asthma,” seeks broad-scale implementation of our effective school-based approach to improve asthma disparities for children in low-income communities (ages 5 to 12 years). Using community-based participatory methods with high risk populations and communities, we will contextualize dissemination and implementation (D&I) of our Colorado school-based asthma program (Col-SBAP) that reduces asthma exacerbations and missed school days, while also addressing social determinants of health (SDOH) which our pilot identified as major drivers of asthma disparities. We will evaluate key metrics identified by diverse stakeholders during a dissemination trial in rural and small metropolitan areas of Colorado. Our dissemination trial will include two interventions: our evidence-based Col-SBAP and an evidence-based assessment and management approach to key SDOH. Our pilot data show both interventions are critical to improve asthma disparities for children from low income families. This two-step level of intervention, called Stop Asthma Attacks (SAA), will be designed for sustainable delivery by school asthma navigators and school nurses who coordinate with primary care and community resources. In partnership with our community stakeholders, the Exploration, Preparation, Implementation, Sustainment (EPIS) D&I framework will be applied during the UG3 planning phase to iteratively adapt our current implementation guide to tailored implementation plans that meet local community and site-specific needs, resources and priorities (EPIS Phases 1 & 2). We will then conduct a UH3 dissemination trial with a randomized stepped-wedge study design in asthma “hot spot” school systems across five Colorado regions to evaluate implementation strategies across asthma “hot spot” school systems (EPIS phase 3, UH3 trial) and to apply the work from EPIS phases 1-3 to develop a “dissemination playbook” to guide future program adoption by other school systems (EPIS Phase 4). The central hypothesis is that SAA will have broad and equitable Reach (primary outcome), and yield important benefits in reducing asthma attacks and symptoms, as compared to schools that have not yet implemented SAA. The SAA playbook will be designed to promote the sustainable adoption of SAA, including training materials and a calculation of return on investment. Our proposal is responsive to and addresses key elements of NHLBI HL-20-003 by “testing late-stage (T4) implementation research strategies” and “promoting and improving population health in high- burden communities” by using a public health approach to target “hot spot” schools with high levels of uncontrolled asthma and asthma associated burden. This proposal will both include a rigorous stakeholder engagement process to ensure SAA is disseminated to diverse geopolitical areas of Colorado with attention to sustainability. Collectively, this evaluation and playbook will accelerate dissemination of SAA nationally to communities experiencing health inequities in pediatric asthma care.

NIH Research Projects · FY 2025 · 2020-09

Summary This proposal is focused on the development of improved therapies for acute myeloid leukemia (AML). The central premise of our all our work is that AML is driven by a biologically distinct leukemia stem cell (LSC) population. While the conceptual importance of targeting leukemic disease at its root is clear, studies in recent years have demonstrated that the inherent intra-patient heterogeneity of LSC populations makes complete eradication a very challenging objective for most patients. Our studies have therefore attempted to identify common foundational properties of primary human LSCs that can employed in the development of therapeutic strategies in the hope that intrinsic heterogeneity can be overcome. Of particular interest, we have described distinct metabolic properties in LSCs, that provide new opportunities for intervention. Specifically, inhibition of BCL2 acts to inhibit oxidative phosphorylation in LSCs, resulting in selective eradication of the LSC population. Recent translation of this observation to clinical studies has demonstrated strong efficacy for newly diagnosed AML patients, and appears to be on the verge of altering the current standard of care. Despite these exciting advances though, relapse remains common and further elucidation of LSC properties is essential. To this end, we have recently begun to describe the mechanisms that drive relapse of AML patients following treatment with a BCL2 inhibitor. These studies have identified entirely unexpected and new aspects of LSC biology that have important ramifications for our basic understanding of AML, as well as the design of improved therapeutic regimens. Specifically, we have demonstrated that at least two distinct LSC populations can co-exist in the same patient. The genetic, epigenetic, and metabolic properties of co-resident LSC subpopulations can vary, giving rise to differing levels of drug responsiveness. The focus of our studies going forward will be to understand and exploit these findings towards the goal of improved outcomes for AML patients.

NIH Research Projects · FY 2024 · 2020-09

Abstract Publiclyavailable genetic summary data canhave high utility for providing insight into genetic etiology of health and disease. Databases of genotype frequencies, such as the genome Aggregation Database (gnomAD), are used to prioritize putative causal variants and, more recently, as pseudo-controls in case-control analysis. Genome Wide Association Study (GWAS) test statistics are used in a variety of secondary data analyses including polygenic risk scores (PRS), genetic correlation analysis, and fine mapping of causal variants. Compared with individual level data, genetic summary data often has fewer barriers in access, promoting broad use of these valuable data resources. The availability and use of summary genetic data is often not equitable across all ancestral groups, especially for understudied ancestral groups that have little to no representation within these resources. Furthermore, heterogeneity within the summary data can lead to confounding and reduced power for case-control analysis, incorrect prioritization of putative causal variants for rare diseases, and reduced accuracy for polygenic risk scores. I develop robust and efficient methods to appropriately use genetic summary data while estimating, modeling, and harnessing the heterogeneity within. My methods coalesce around a unifying framework where I flip the paradigm of genetic and genomic data treating the genetic variant or element as the observational unit by which we analyze the data rather than the individual. This simple, yet innovative paradigm shift enables the use of classical statistical techniques and the creation of methods that detect, adjust for, and even use heterogeneity within summary level data. To enable broad and equitable use of our methods, we will create publicly available R packages compatible with Bioconductor and Shiny Apps for interactive internet use.

NIH Research Projects · FY 2024 · 2020-09

Tuberous Sclerosis Complex (TSC) is a multi-organ disorder caused by mutations in the TSC1 or TSC2 genes. TSC is a challenging disease to approach as there are many involved organ systems, which have distinct profiles of symptom onset, disease progression, and in some cases even stability or regression of the benign tumors known as hamartomas. These multiple examples of distinct time courses in each organ system strongly suggest that the TSC1/TSC2 genes control cell signaling pathways that are tissue specific and developmentally regulated, resulting in lesions that present at different times in the lifetime of the patient. These pathways certainly include mTOR kinase signaling, but critical upstream and downstream regulators of this developmentally regulated process in specific tissues remain poorly understood. The neurological manifestations of TSC are typically severe at very early ages and include epilepsy, intellectual disability, autism, and behavioral/psychiatric disorders. Recent findings in human cell-derived model systems suggest that neural development is disrupted in TSC, and that proper regulation of mTOR signaling is especially important in human brain in comparison to other mammals. However, the cellular mechanisms connecting TSC1/2 mutation and the phenotypic outcomes of this mutation are not well understood. This project will use patient-derived cells and single-cell measurements of protein and RNA to measure altered signaling pathways in various cell types of the human brain and also address how these abnormalities impact specific developmental stages. Examined stages will span early neural progenitor cells to more mature neurons found in the postnatal brain. Human induced pluripotent stem cells (iPSCs) from patients carrying TSC2 mutations will be used to generate lineage-committed progenitors and differentiated neurons and glia. We will also use freshly resected human tubers as well as previously resected human tubers that have been fixed and stored, and will employ custom-designed computational pipelines to compare the developmental trajectories of TSC2-mutant cells to matched controls and larger published datasets. Using cutting edge cell imaging and analysis protocols, we will test the overarching hypothesis that tubers from patients with TSC and stem cell derivative neural cells and tissues have mTOR-dependent and mTOR- independent signaling abnormalities that are lineage- and temporally-restricted. Finally, we will quantitatively compare signaling dynamics in specific developmental stages and lineages between TSC2 mutant cells and cells derived from a second “mTORopathy” with overlapping but non-identical clinical features, to dissect the function of different components of this pathway in neural development and pathogenesis and reveal compensatory signaling after treatment of cells carrying TSC2 or DEPDC5 mutations.

NIH Research Projects · FY 2024 · 2020-09

Project Summary While much research has been devoted to exploring the impact of environmental chemical exposures during pregnancy on infant and child health, relatively little attention has focused on the potential influence of these exposures on maternal health. Recent evidence suggests that pregnancy may be a sensitive period in the life course, during which chemical exposures may have long-lasting effects on cardio-metabolic disease risk among women. Using a well-characterized existing cohort study that enrolled 1,410 pregnant women in 2009- 2014, we propose the following aims: (1) quantify the relationship between environmental exposures during pregnancy and short-term maternal health outcomes including: postpartum weight retention, reduced breastfeeding initiation and duration, and incident diabetes; (2) quantify the relationship between environmental exposures during pregnancy and long-term maternal health outcomes: body composition, weight trajectories from pregnancy through ~10 years after parturition, hepatic fat, dysglycemia and incident diabetes, and cardiovascular disease; and (3) evaluate the potential role of maternal characteristics and behaviors during pregnancy, specifically obesity and diet quality, in modifying associations between environmental chemical exposures and outcomes. Exposures during pregnancy will include serum per- and polyfluoroalkyl substances (PFAS), urinary phthalate metabolites, phenols and parabens, metals, organophosphate flame retardants, and modeled air pollutants at the maternal residential address during pregnancy. We propose to recruit 700 of the original study participants to return for a follow-up visit at ~10 years postpartum. At this visit, participants will undergo a comprehensive metabolic health evaluation including body composition via air displacement plethysmography (BOD POD), dysglycemia via oral glucose tolerance test, and hepatic fat fraction via MRI. Medical records will be abstracted to document incident diabetes and cardiovascular disease, and to reconstruct body weight trajectories. We will estimate associations between exposures during pregnancy and maternal outcomes using covariate-adjusted multivariable regression models for continuous, binary, or time-to- event data, as appropriate. Exposures will be evaluated as single pollutants and as mixtures using advanced statistical methods including Bayesian Kernel Machine Regression and Bayesian hierarchical Cox survival models. We hypothesize that maternal body mass index prior to pregnancy and diet quality during pregnancy will modify the effects of environmental chemical exposures on cardio-metabolic outcomes, such that associations will be stronger among women with obesity entering pregnancy or with poor diet quality during pregnancy. The results of this study will inform public health interventions to identify women who may be especially susceptible to the effects of environmental chemical exposures during pregnancy, and to improve the environment of pregnancy to promote the long-term health of both the offspring and the mother.

NIH Research Projects · FY 2024 · 2020-09

Summary Pharmacogenetic testing is expected to enhance drug safety and efficacy while improving clinical outcomes by tailoring treatment regimens to the individual. Key to maximizing benefits of this approach will be comprehensive characterization of genetic variation that modulates drug absorption, distribution, metabolism and excretion, drug target interaction and pharmacological response across populations. Significant advances in cataloguing pharmacogenomic profiles from major populations of the world have aided clinical applications but have largely excluded historically marginalized populations and small, geographically isolated populations, including American Indian and Alaska Native (AIAN) people. These populations continue to be underrepresented in genomic research for a variety of reasons: lack of community engagement, historical mistrust, and geographical remoteness. Recent genetic studies by the Northwest Alaska Pharmacogenetic Research Network (NWA-PGRN)—a center with multiple institutional and tribal partners—have shown that AIAN people carry variants in pharmacogenes that are both novel and common, have allele frequencies of known variants that are different than other ethnic or racial groups, and display high inter-tribal variability as well. To broaden this research, we propose to leverage the lessons learned and build on research from the NWA-PGRN to understand inter-individual variation in drug responses and provide novel indicators and guidelines for implementing personalized medicine in AIAN communities. Our overall goal is to fill this knowledge gap through characterizing genomic variation of pharmacogenes in a wide range of AIAN populations using an ethical and community-engaged framework that focuses on developing and deepening research partnerships with AIAN communities. We also seek to investigate the relationships between genotype-phenotype of novel variants using in silico prediction and in vivo functional assays. This work will create a general model for genomic research that engages communities in the research plan throughout the research project (from start to completion and beyond) and will serve as a foundational way to shift the research framework in genomics.

NIH Research Projects · FY 2025 · 2020-08

ABSTRACT FDR from FIP Family Our proposed rare disease cohort focuses on a critical unmet public health need in (N=1000) interstitial pneumonia, to understand the etiology, natural history, and phenotypic N=650 Subcohort N=350 heterogeneity of preclinical pulmonary fibrosis (PrePF), before the lung is scarred irreversibly. Our overall hypothesis is that common genetic variants and 100 PrePF 50 PrePF environmental risk factors predispose to the development, natural history, 300 unaffected and phenotypic heterogeneity of PrePF, and that defining these risk factors will allow us to uncover the common and unique subtypes of PrePF that differ Case-Cohort at baseline in disease onset and progression. By leveraging our NHLBI-supported (150 Cases; 300 Unaffecteds) discoveries in PrePF, familial interstitial pneumonia (FIP), and idiopathic interstitial Figure 1. Relationship between pneumonia (IIP), and our NHLBI-supported cohort of FIP families, our proposal Subcohort and the original FIP seeks to explain how common genetic variants and the environment interact to cohort. Given the risk of PrePF FIP FDRs (15%), we anticipate result in the earliest stages of this highly morbid, phenotypically heterogeneous that 50 individuals in the disease. We will focus on first-degree relatives (FDRs) previously phenotyped as Subcohort will have PrePF at unaffected (N=2404) from our 1160 FIP families with two or more cases of baseline. Cases will be supplemented from the confirmed IIP. Within these 1160 FIP families, we will establish our rare disease remaining 650 subjects in the cohort of 1000 subjects by selecting up to two asymptomatic, previously phenotyped overall cohort so that our case- unaffected FDRs per family. To combine the advantages of our cohort with the cohort population at baseline should include 150 cases of efficiency of a nested case-control study, we will establish a case-cohort study and PrePF and 300 unaffecteds. compare 150 cases of PrePF to 300 unaffected controls (Figure 1). This approach will allow us to determine the individual and combined contributions of common genetic variants and environmental features that result in the development of PrePF. By focusing on the natural history of IIP, our results can be used to identify high-risk populations, early forms of the disease, factors associated with disease progression, and biological targets for drug development. Moreover, a natural history study can also identify critical biomarkers that can be diagnostic of early or established forms of the disease, prognostic of the course of a disease, predictive of treatment response, or useful in guiding patient selection and dose selection in drug development programs. Our proposal would establish a prospectively followed high-risk IIP cohort, will identify the genetic and environmental risk factors for PrePF, and will maximize the utility of this high-risk cohort for ancillary studies focused on primary and secondary prevention of IIP.

NIH Research Projects · FY 2024 · 2020-08

PROJECT SUMMARY/ABSTRACT Candidate: Dr. Natalie Nokoff is a Pediatric Endocrinologist whose research and clinical focus is on the cardiometabolic health of transgender youth. The goal of her K23 proposal is to acquire the critical research skills necessary to lead high-quality, patient-centered clinical research. This training will help her achieve her long-term goal of developing a successful independent research program and becoming an international expert on the impact of sex steroids on cardiometabolic health. Background: Up to 1.8% of adolescents in the U.S. identify as transgender. Treatment of transgender females (male sex and a female gender identity, MTF) may include pubertal blockade (gonadotropin-releasing hormone analogue or GnRHa) in early puberty followed by estradiol later in adolescence. The impact of these treatments on cardiometabolic health is understudied and unclear. MTF adults on estradiol have higher rates of vascular events than non-transgender (cisgender) individuals. Research Plan: Pubertal Blockade and Estradiol Effects on Cardiometabolic Health for Transitioning Youth (PUBERTY) is a longitudinal observational study to evaluate the impact of GnRHa and estradiol on vascular function and insulin sensitivity (IS). Young MTF (n=15) will be evaluated before and 6 months after GnRHa. A separate cohort of MTF youth (n=40, ages 13-16 years, 20 who clinically received a GnRHa and 20 who did not) will be evaluated at baseline, 6 and 12 months after estradiol. MTF youth will be compared to cisgender youth (n=15 males, 15 females), adjusted to pubertal stage and body mass. Career Development Plan: Dr. Nokoff’s research career development training activities include: 1) gain expertise in design and conduct of clinical studies (implementation and completion of the PUBERTY study, coursework and a PhD in Clinical Investigation); 2) lean how to perform and interpret vascular studies; 3) learn new methods for assessing IS. Environment: The environment for this project is exceptional with a strong academic Section of Pediatric Endocrinology at the University of Colorado Anschutz Medical Campus and resources of the Colorado Clinical and Translational Sciences Institute and Center for Women’s Health Research. Dr. Nokoff is faculty in the TRUE Center for Gender Diversity, a comprehensive, multidisciplinary clinic serving over 1,000 transgender youth, that will assure successful recruitment for this study. Mentors with expertise in endocrinology, insulin resistance, vascular health, sex steroids, working with vulnerable populations, and clinical investigation are invested in this candidate’s future and will be instrumental in supporting her research and career development. Impact: The results of this novel study will improve the clinical care of transgender youth and yield information on risk factors for future vascular events. Dr. Nokoff will gain invaluable skills in study design as well as assessment of vascular and metabolic health, launching an impactful career as a lead clinical investigator.

NIH Research Projects · FY 2025 · 2020-08

Project Summary/Abstract The problems of obesity and type 2 diabetes (T2DM) are some of the most common conditions affecting Americans. There is a critical need to train the next generation of MD/DO and PhD investigators who will address these public health challenges with innovations in basic, clinical and translational sciences. We believe that the University of Colorado, Anschutz Medical Campus (UCAMC) is well positioned to provide the training needed to produce the next generation of scientists addressing these areas. We are seeking continuing support for 4 positions (2 post-doctoral trainees to be supported for 2 years each) in the Research Training Program in Metabolism, Obesity and T2DM (RTPMOD) at the UCAMC. The strength of the RTPMOD lies the outstanding faculty, state of the art facilities and programs at the UCAMC and a robust pipeline of applicants. The 25 members of the Training Faculty are productive well-funded investigators, many of whom are nationally recognized leaders in their fields of study who collectively have $16.5 million in current research funding (direct costs current year). The UCAMC is host to the Colorado Clinical Translational Science Institute, the Colorado Nutrition Obesity Research Center, the Denver Diabetes Research Center and the Anschutz Health & Wellness Center to name just a few of the resources that will support the requested training. Over the last 4 years of this first cycle of the new RTPMOD, 8 postdoctoral fellows have been supported. Two received F32 awards and 2 received K awards. All remain in research related or research-intensive positions. Two are Assistant Professors. Applicants for the RTPMOD come from a range of clinical fellowship programs at UCAMC, our clinical Obesity Medicine Fellowship, and the Physician Scientist Training Program track within our Internal Medicine Training Program. PhD applicants come from local and national recruitment efforts. MD/DO and PhD applicants are selected though a rigorous recruitment process designed to identify those with the greatest promise to pursue an independent research career. In addition to an intensive mentored research experience, trainees will gain formal training in metabolism, obesity and T2DM and team science. The program will be administered by an experienced Program Director supported by an Executive Committee with input from a independent External Advisory Board. We believe that the RTPMOD has shown early success in this first cycle of funding and provides outstanding training for postdoctoral fellows seeking a long-term independent research career in metabolism, obesity or T2DM research.

NIH Research Projects · FY 2024 · 2020-08

Project Summary/Abstract This is a first submission for an NHLBI K23 Mentored Patient-Centered Career Development Award (PA-19- 119) by Christopher Knoepke, PhD, MSW, LCSW. Dr. Knoepke’s goal is to be a leading social work scientist improving the methods by which patients’ values are included as they consider advanced therapies. This proposal uses Transcatheter Aortic Valve Replacement (TAVR) as a model for studying decision support for cardiovascular disease patients generally. Candidate: Dr. Knoepke is a Licensed Clinical Social Worker who is currently an Assistant Research Professor in the Division of Cardiology at the University of Colorado School of Medicine. He has a Masters (Washington U. in St. Louis) and PhD in social work (U. of Denver). He works closely with his primary mentors (Drs. Daniel Matlock, MD, MPH & Larry Allen, MD, MHS), receiving mentorship from mentors/collaborators (including Drs. Russ Glasgow, Megan Morris, Eric Campbell, John Carroll, and Laura Scherer) through the Colorado Cardiovascular Outcomes Research Group (CCOR) & the University of Colorado’s Dissemination & Implementation Workgroup, and ACCORDS. Training/Mentors: This award augments Dr. Knoepke’s earlier training by focusing on three areas relevant to patient values in decision support:1) Advanced qualitative research; 2) clinician survey methodology; and 3) Dissemination & Implementation science. Dr. Knoepke proposes intensive mentorship; coursework & workshops, and contextual learning through the proposed research. Drs. Knoepke and Matlock have assembled a strong team of local mentors and collaborators to guide Dr. Knoepke through the training and research activities. These activities will enable Dr. Knoepke to innovatively develop decision support tools for real world care. Research: There is a pressing need to improve the science of clarifying implementing the assessment of patient values as they decide whether to accept advanced therapies. Advanced CVD patients are disproportionately offered such therapies. As advanced age & comorbidity can reduce expected benefits, the need to include patients’ values in decisions is paramount. This study will produce information relevant to clinical practice. Aim 1 will characterize the current state of practice in values clarification as it actually occurs between patients and care providers when they discuss TAVR. Aim 2 will include the development and deployment of a de novo survey of cardiology specialists, assessing design and contextual factors necessary to support dissemination of shared decision making in TAVR. Aim 3 will pragmatically pilot adapt a decision aid for older and/or high risk TAVR candidates (separately developed by Dr. Knoepke) to include findings from Aims 1 & 2. Summary: This award’s innovative collaboration between social work and cardiology will maximize implementation of findings and facilitate Dr. Knoepke’s development into a leader in improving how patient values are honored as they make complicated decisions about their health.

NIH Research Projects · FY 2024 · 2020-08

PROJECT SUMMARY Fibrotic disorders account for a significant source of global morbidity and mortality. Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and life-threatening lung disease most prevalent in elderly populations. IPF impacts 100,000 patients in the U.S. alone and there are approximately 34,000 new global diagnoses each year. Most patients with IPF succumb to respiratory failure within 3-5 years and the only clinically available therapeutic treatments do not cure the disease. As the average age of the U.S. population increases, it is imperative for researchers and practitioners to work together to identify new targets to halt or reverse IPF. Discovery of new therapeutic targets for IPF through traditional cell culture techniques and pre-clinical animal models has several limitations because these systems do not adequately reproduce key aspects of human physiology. Most importantly, dynamic cell-matrix and cell-cell interactions that are difficult to recapitulate in vitro drive the progression of fibrosis: it is not clear, for example, whether changes in the extracellular matrix (ECM) composition or the subsequent alterations in mechanical properties of the surrounding tissues are the more potent drivers of IPF, i.e., the best target for therapeutics. New tools and technologies that enable us to dynamically study the pathogenesis of fibrosis over time remain an unresolved challenge. My laboratory has developed novel methods to synthesize and microfabricate a new class of biomaterials to conduct dynamic cell-ECM studies, not currently possible in traditional models of fibrosis. Our innovative platform combines a phototunable poly(ethylene glycol) (PEG) backbone with clickable decellularized ECM (dECM) from healthy or diseased lung tissue so that we may decouple fibrotic tissue composition (e.g., increased collagen content) from subsequent changes in mechanical properties (e.g., increased stiffness). Specifically, healthy or IPF lung dECM will be incorporated into soft (1-5 kPa) hydrogel matrices that mimic healthy tissue, then exposure to focused light will dynamically initiate stiffening to fibrotic levels (>10 kPa). Three aims are proposed to engineer and implement this biomaterials-based strategy for building novel, high- fidelity in vitro models of IPF. AIM I: Engineer the structure, composition, and dynamic mechanics of PEG- dECM cell culture platforms to recapitulate distal lung tissue; AIM II: Interrogate the impact of composition and mechanical properties on fibroblast activation using dynamic PEG-dECM biomaterial platforms; and AIM III: Identify druggable mechanosensitive targets of the fibrotic activity recreated in dynamic 3D models. Successful completion of these aims will advance our understanding of the cellular and molecular drivers of IPF, building the foundation for high-throughput discovery and screening of therapeutics for precision medical treatments.

NIH Research Projects · FY 2026 · 2020-08

Project Summary Ion channels are precisely tuned molecular machines that control the flow of ions across the plasma membrane. They can sense a wide array of stimuli including voltage, pH, neurotransmitters, temperature, and mechanical force. The function of these proteins depends critically on an array of factors that can regulate their activity including the composition of the lipid membrane, protein interacting partners, post-translational modifications, and assembly with different subunits. Our lab focuses on two families of ion channels: Acid-sensing ion channels (ASICs) and hyperpolarization-activated cyclic nucleotide-gated (HCN) channels. Acid-sensing ion channels (ASICs) are critical sensors of extracellular pH that contribute to excitability in cells in both the central and peripheral nervous system. ASICs are trimeric sodium selective ion channels that can assemble as both homo- and heterotrimers and their precise properties are governed by the channel composition. They have important roles in cell death post ischemia as well as in pain sensing. HCN channels are unusual voltage-gated channels that respond to cellular hyperpolarization and the direct binding of the cyclic nucleotide cAMP. HCN channels are important determinants of cellular excitability in a neurons and cardiac pacemaker cells. Both families of channels have emerged as potential drug targets for pathophysiological conditions like stroke and pain. Our research seeks to understand mechanisms of channel function and regulation for both families of channels. This grant outlines three ongoing lines of research being conducted in the lab. 1) We are seeking to understand the structural determinants of lipid regulation of ASIC3. We have shown that ASIC3 is potentiated by at least three classes of single acyl chain lipids: polyunsaturated fatty acids (PUFAs), lysophosphatidyl cholines (LPC), and N- acyl amino acids (NAAAs). The effects of these lipids can be strong enough to activate the channel in the absence of acidification suggesting ASIC3 may also serve as a lipid sensor. Our goal is to understand how lipids bind to ASIC3 and how those interactions alter channel function. 2) We have developed a novel single molecule photobleaching technique to determine ion channel stoichiometry. We are using this new approach to look at how ASICs that are expressed in dorsal root ganglion cells (ASIC1a, ASIC2b, ASIC3) assemble into heteromeric channels. 3) Finally, we are studying how unique structural elements in HCN4 make it particularly sensitive to modulation by ions, small molecules, and protein binding partners. The site of this isoform specific difference in HCN4 is likely the C-linker which couples the cyclic nucleotide binding domain to the pore of the channel. By understanding the sites of regulation for both families of channels we believe we can uncover novel sites for the development of drugs that target each of these families of ion channels.

NIH Research Projects · FY 2024 · 2020-08

ABSTRACT Our long-term goal is to generate a complete understanding of how the cerebellum learns to improve movement in response to motor errors. Climbing fibers are thought to play an essential role in this process because they fire during erroneous movement. Their activity reliably excites Purkinje cells, eliciting calcium spikes in their dendrites that can trigger long-term synaptic plasticity at coactive parallel fiber inputs. Plasticity induction ultimately leads to corrective behavior by altering the cerebellum’s response to sensorimotor stimuli that predict mistakes. Importantly, inhibition from molecular layer interneurons (MLIs) that target Purkinje cell dendrites suppresses climbing-fiber-evoked calcium signaling, opposing or ‘gating’ plasticity induction. Because MLIs are activated by movement, this suggests Purkinje cell disinhibition is required during motor learning. As MLIs inhibit other MLIs, their interconnections could support a circuit for Purkinje cell disinhibition during behavior. The objective of this proposal is to examine the possibility that MLI circuits are structured to support a context- dependent engagement that allows climbing fibers to instruct plasticity and learning in response to motor errors. To accomplish this, we will employ a multidisciplinary approach using cutting-edge molecular-genetic techniques, functional recordings, circuit mapping, and behavioral analysis. In the first aim, we will test whether ablating MLI- to-MLI connections that normally support Purkinje cell disinhibition affect the ability of climbing fibers to evoke full-blown calcium signals in response to motor errors, and whether loss of MLI-MLI circuit function affects cerebellar-dependent motor learning. In the second aim, we will establish an MLI taxonomy and use it to survey for previously unknown MLI subtypes. We will also use functional recordings to test whether there is evidence for bias connectivity within the MLI network that supports a dedicated circuit for Purkinje cell disinhibition. In the third aim, we will use anatomical tracing to ascertain the MLI connectome. In this way we will determine if there is a structural basis for the independent actuation of MLI subtypes through their afferent inputs and the cell-type selectivity of their efferent outputs. Completion of these aims will lead to an unprecedented understanding of the organizational logic of the molecular layer. In particular, we expect to reveal how circuits within the molecular layer control the induction of climbing-fiber-mediated learning. This knowledge will not only help develop theories/models of cerebellum function but will also provide insight into the processes underlying learning in general.

NIH Research Projects · FY 2024 · 2020-08

Pulmonary arterial hypertension (PAH) afflicts patients of both sexes and across a broad age range and is highly lethal, if not promptly diagnosed and appropriately treated. Despite advances in the understanding of its pathogenesis and the development of 14 therapies approved by the United States Food and Drug Administration (FDA) over the past 2-3 decades, it continues to be associated with significant morbidity and mortality. The fundamental premise in this PPG is that PAH is highly heterogeneous regarding clinical parameters including initiating factors, clinical presentation, rate of progression, and response to therapy. Importantly, patient-to- patient heterogeneity, involving the types of pulmonary vascular lesions and the corresponding endotypes (i.e., underlying molecular processes driving the specific disease presentation), has been uncovered by our group and underlies the high complexity of disease pathogenesis. The paucity of investigations of these broader aspects of heterogeneity has resulted in a lack of understanding of specific factors contributing to particular sub- phenotypes of PAH – negatively impacting the development of more targeted (and individualized) therapies. These limitations manifest in the fact that some patients live many years on currently available treatments – however without cure of their disease -, while others progress rapidly and inexorably from onset to transplantation or death. This proposal seeks to uncover novel pathogenetic processes linking pulmonary vascular inflammation, remodeling, and molecular underpinnings of pulmonary vascular lesions in PH, while recognizing the key pathological and pathobiological heterogeneity of the disease. The central premise to be tested in this proposal is that early and persistent local, pulmonary vascular-specific activation of complement leads to persistent perivascular inflammation and extracellular matrix changes, thus shaping a feed-forward loop of pro- inflammatory/pro-remodeling perivascular microenvironment, leading to development of PH. This application represents a major step towards identifying new biomarkers and more effective individualized treatments, which are critically needed.

NIH Research Projects · FY 2024 · 2020-08

Cognitive decline majorly affects quality of life in the general aging population; this is further exacerbated by an increased risk for neurodegenerative diseases. The general age-related cognitive decline is thought to be mainly due to impaired synaptic function, not loss of neurons. Similarly, while neurodegenerative diseases do involve loss of neurons, there is also significantly impaired synaptic function in the surviving neurons, For instance, amyloid β oligomers (Aβ) are major pathological agents in as Alzheimer's disease (AD) and cause acute impairments in long-term potentiation (LTP) of excitatory synapses in the hippocampus, even at time points and concentrations insufficient to induce any significant neuronal cell death. Here we will test our hypotheses that the LTP impairments related to normal aging versus AD (i) both involve mis-regulation of the Ca2+/calmodulin(CaM)-dependent protein kinase II (CaMKII), but (ii) by fundamentally different mechanisms to (iii) result in the distinct forms of LTP impairment in normal aging versus AD. Specifically, we hypothesize that CaMKII hypo-nitrosylation directly causes the impairments in aging, but not the Aβ-induced impairments (which may instead even involve hyper-nitrosylation). Additionally, we hypothesize that hypo-nitrosylation reduces LTP by chronic long-term effects on synapse composition (including CaMKII itself), while the Aβ effects instead involve acute mis-regulation of CaMKII. LTP is well-known to require CaMKII and its Ca2+-independent “autonomous” activity that is generated by autophosphorylation of T286. Additionally, two alternative ways to generate autonomous activity have been described by my lab: Binding to the NMDA-receptor subunit GluN2B and S-nitrosylation of C280+C289. Indeed, CaMKII binding to GluN2B is also required for normal LTP and for the CaMKII movement to excitatory synapses during LTP. The functions of CaMKII nitrosylation in LTP and other forms of synaptic plasticity will be elucidated here. Intriguingly, previous studies have shown that aging is accompanied by hypo-nitrosylation of neuronal proteins, including CaMKII, in both mice and humans. Additionally, preliminary studies indicated that nitrosylation causes CaMKII movement to excitatory synapses, and that this requires regulated CaMKII binding to GluN2B. i.e. the same mechanism that is required for the LTP-induced CaMKII movement. In three related but independent aims, our proposal will determine the specific involvement of CaMKII nitrosylation in the LTP impairments related to normal aging versus AD (with the expectation for fundamentally distinct CaMKII mis-regulation). First, we will determine the regulatory mechanisms for synaptic CaMKII localization by nitrosylation. Then, we will determine the functions of CaMKII nitrosylation in the distinct impairment of LTP related to normal aging versus AD. Finally, we will determine the effects of CaMKII nitrosylation on learning and memory function in behavioral tasks.

NIH Research Projects · FY 2025 · 2020-08

Project Summary The Rocky Mountain Summer Research Education Experience (RMSREE) Program will fill a critical need to increase the STEM knowledge and understanding of Denver area high school teachers, enhancing the STEM content in the K-12 educational agencies in which they teach and ultimately resulting in increased participation and better training of the biomedical sciences research workforce. The main goals of the RMSREE program are: 1) to provide substantive research training, individualized academic preparation, and career guidance to strongly enhance the skills of the RMSREE teacher participants, Research Education Fellows; 2) to recruit a diverse set of Fellows who are committed to translating their research experience into enhanced STEM education in their classrooms and provide them with professional development opportunities to effectively bridge the gap between the research setting and the HS STEM classroom; and 3) to continue to create and sustain long-term connections between CU Denver and local high schools that will improve the STEM education of HS teachers as well as provide a STEM pipeline for their students to enter into the biomedical imaging, bioengineering, and health informatics professions. The RMSREE program holds high expectations for performance (95% of participants adding significant STEM content by the end of their first year and 80% maintaining that level over the five-year period following the program). To achieve these ambitious outcomes we will recruit cohorts of highly-committed, ambitious candidates and engage them in an integrated set of activities, centered around meaningful research experiences, but also augmented with mentoring through individualized feedback on professional development plans (PDPs), a weekly seminar series, and training in laboratory culture and the responsible conduct of research. This will build participants' research skills and STEM content knowledge and understanding as well as promote their progression to become more effective STEM teachers. Group activities will contribute to building a cadre of trainees who will continue to support each other throughout their careers to optimize professional success. The RMSREE program will serve six Fellows each year, for a total of thirty trainees over the 5-year project period, which will in turn impact many hundreds of their students in the years following the project.

NIH Research Projects · FY 2026 · 2020-07

Epidemics of chronic kidney disease of unknown etiology (CKDu) have emerged in Central America (Mesoamerican Nephropathy), in northern Sri Lanka (Sri Lankan nephropathy), in Andhra Pradesh and other regions of India (Uddanam Nephropathy), in Veracruz and Aguascalientes regions of Mexico and are emerging in the United States. In all cases, the primary histologic finding is chronic interstitial nephritis with variable degrees of glomerulosclerosis. To date a variety of causes have been considered, including heavy metals, agrochemicals, infectious diseases, and recurrent heat stress and dehydration. During the first cycle of this award, we evaluated the novel hypothesis that amorphous silica, released into the air during the burning of sugarcane and rice, may be a primary cause. Our findings demonstrated that: 1). Amorphous silica present in sugarcane and rice and is released when crops are burned as part of harvesting. 2). The particulate matter fraction (PM2.5) of burned sugarcane and rice contains 80-90 percent amorphous, nanoparticle sized silica. Workers at risk for CKDu are exposed to extremely high air borne concentrations 3). We reported that individuals with CKDu have higher levels of silica nanoparticles present in kidney biopsies than those with other kidney diseases 4). Sugarcane workers have increasing levels of silica in their urine across a harvest season. 5). Inhalation of amorphous nanoparticle sized silica or sugarcane ash in rats or mice leads to development of CKDu like pathology with minimal respiratory signs and the biopsies show chronic interstitial nephritis with silica particles present in kidney. Given our important findings during the first cycle of this award, we propose to 1) Characterize the exposure to silica in a cohort of sugarcane workers in Guatemala and examine levels of silica in urine, blood and nasal washes. 2) Develop a novel physiologically based toxicokinetic model in mice that can predict exposure to silica in humans and be extrapolated to workers at risk for CKDu. 3) better determine mechanisms of toxicity and identify novel biomarkers by using an integrated approach of redox proteomics and spatial transcriptomics in an animal model. Overall, these studies will improve our understanding of the role silica exposure has on development of CKDu, develop a novel predictive model to better determine risk associated with exposure and greatly improve environmental safety and human health.

NIH Research Projects · FY 2025 · 2020-07

PROJECT SUMMARY Diabetes is a global heath epidemic affecting >500M people worldwide. Even with rigorous management increased risk for diabetic complications remains. Bioengineering approaches are providing innovate technologies that are overcoming the current limitations in diagnostics, treatments and cures for diabetes and complications. However, there is a shortage of a skilled scientific workforce able to work on these developments. The University of Colorado is uniquely capable to host this innovative program - Bioengineering Interdisciplinary Training for Diabetes Research - to train future scientists with the skills and knowledge to solve existing and future problems in diabetes research and clinical care. This includes worldrenowned research and clinical programs in diabetes; exceptional engineering research and training programs; and bioengineering research and training that is fully integrated with the Medical campus. The primary goals for the next phase of this program are: 1 ). To attract high-quality trainees with engineering/quantitative backgrounds; 2). To provide in-depth, multi-disciplinary research training for predoctoral trainees integrating bioengineering and diabetes; and 3). To prepare trainees for transition to individual fellowships and research careers in academia or industry. To achieve these goals, our research training follows 3 core principles: a). That a successful bioengineering career requires an intrinsically multidisciplinary approach, with scientists equipped with skills covering a broad variety of fields; b). That successful training of these scientists will require constant interactions between trainees and crossdisciplinary mentors at all levels of research; and c). That developing such scientists requires an institutional environment with breadth and depth, established interactions between engineering and clinical science, and faculty committed to this training. This innovative research training program combines didactic and hands on coursework, lab-based research, clinical experiences and career skills. Research areas focus on true strengths at the institution in which established interactions between bioengineering and diabetes are present, including biomarkers and therapeutic interventions for diabetes and complications; immune engineering; clinical trials; imaging diagnostics for diabetes and complications; and computational methods for diabetes management and artificial pancreas. We achieved excellent outcomes during the previous funding period, including recruiting high quality trainees; trainees publishing extensively and receiving fellowships; and graduating and continuing into research careers. These outcomes; the depth of expertise in diabetes and engineering research; the significant institutional support; and the extensive facilities available make a compelling argument to continue to fund such a program at the University of Colorado.

NIH Research Projects · FY 2025 · 2020-07

Project Summary / Abstract Tumorigenesis requires cells to bypass or escape two discrete and distinctive anti-proliferative barriers: replicative senescence and crisis. Senescence is a permanent cell cycle arrest, activated as a primary response to telomere deprotection and involves stimulation of the tumor suppressor pathways p53-p21WAF1 and/or p16INK4A- Rb. Disruption of cell-cycle checkpoints renders cells capable of bypassing senescence and continuing proliferation, while telomeres shorten further. Eventually such cells initiate a terminal response called replicative crisis, during which critically short telomeres become subject to end-to-end fusions, resulting in massive cell death. On rare occasion, a small group of cells will emerge spontaneously from crisis and evolve towards malignancy, yet the mechanisms underlying cell death in crisis and crisis escape are not defined. Dr. Joe Nassour has recently discovered an unrecognized function for macroautophagy (hereafter autophagy) in the elimination of cells during crisis. Autophagy is therefore an essential component of the crisis response required for the removal of cells at risk for malignant transformation. This suggests that autophagy defects can be the molecular basis for tumorigenesis. In his Pathway to Independence Award (K99/R00) proposal, Dr. Nassour, together with his Mentor Dr. Jan Karlseder, and his Co-Mentors Dr. Reuben Shaw, Dr. Martin Hetzer, and Dr. Peter Adams, designed a dedicated training plan and proposed a research project that sets out to dissect the molecular basis of mammalian autophagy and its potential therapeutic role in the earliest stages of human cancer. In particular, Dr. Nassour will focus on deciphering the mechanism of autophagy-dependent cell death in crisis (Aim 1), elucidating the interplay between autophagy and genome stability (Aim 2), and evaluating the role of autophagy in neoplastic transformation through crisis escape (Aim 3). The in vivo relevance of Aim 3 will be examined by employing knockout and transgenic mouse models susceptible to telomere dysfunction-driven carcinogenesis. The occurrence of cellular crisis in tissues and the impact of autophagy on tumor incidence will be examined. This research will provide new insights into the function of autophagy in cancer biology, and should provide a rationale for developing autophagy modulation approaches to ameliorate the efficacy of cancer therapy. Dr. Nassour’s training plan will provide all the necessary professional development to direct an independent laboratory using next-generation sequencing (NGS)-based ‘omics’ approaches and transgenic mouse models to define the mechanisms and function of autophagy in cancer. Training modules in this award include: Computational analysis and bioinformatics for NGS data, methods for handling and restraint in the mouse, transgenic mouse technology, and mentorship skills such as teaching and grant writing; which will all be necessary for the success following the transition to independence.