University Of Arizona

NIH Research Projects · FY 2025 · 2016-06

PROJECT SUMMARY: Asthma, chronic obstructive pulmonary disease (COPD) and the Preserved ratio impaired spirometry (PRISm) are associated with increased morbidity and mortality across the lifespan. Recent discoveries strongly suggest that the roots of many cases of these three heterogeneous conditions can be found in early life. Identifying the lifetime molecular pathways underlying them will provide a new understanding of the pathogenic mechanisms involved in their inception. The Tucson Children’s Respiratory Study (TCRS) is a birth cohort that has already made major contributions to our understanding of the natural history of lung function trajectories and the early origins of asthma and COPD. A recent tantalizing result was derived from the use of lung function measurements obtained repeatedly from early childhood through the beginning of the 5th decade of life to model latent class trajectories in over 600 participants. We identified a novel resilient trajectory, in which lung function decline occurs at a slower rate than in their peers. Early data suggests that this resilience may be afforded through increased lung protective antiproteases molecules. We also identified individuals with dysanapsis, i.e., disproportionate scaling of airway dimensions to lung volume, which is a risk factor for COPD. A strong precursor of dysanapsis was excessive weight gain between infancy and the early school years and we found a specific serum biomarker at age 6, PTX2, that may mediate this association. We showed that restrictive spirometry is associated with poor prenatal and early life growth and detected early life developmental modules that may be responsible for these differences. We identified phenotypes related to the natural history of asthma, and linked these to novel metabolic pathways that may be responsible for the onset or persistence of asthma. TCRS participants are in the 5th decade of life, when lung function changes associated with chronic lung disease become increasingly more apparent. In this application, we have adapted new technologies, such as gold standard metabolomics, proteomics, and single-cell and bulk RNA sequencing, applied to longitudinally collected samples, in combination with ongoing assessment of the participants. Using these technologies, we will investigate, prospectively, the early risk factors for these lung disorders at the molecular level. We will address three specific aims: 1. To continue following TCRS participants to further characterize different lung function trajectories and phenotypes from childhood into adult life, and to specifically assess the molecular underpinnings of lung dysanapsis and of the obstructed and resilient trajectories. 2. To identify profiles in the serum proteome and peripheral blood gene expression patterns associated with poor postnatal somatic growth, and to relate these profiles to the restricted spirometric trajectory. 3. To characterize the novel cellular and molecular mechanisms underlying the natural history of asthma. As the only birth cohort with hundreds of non-selected participants followed from birth into the fifth decade of life, the TCRS offers a unique opportunity to investigate the longitudinal mechanisms underlying these lung disorders.

NIH Research Projects · FY 2025 · 2016-05

ABSTRACT Dedicated breast CT is an emerging technology with approximately 20-25 clinical prototypes and clinical systems worldwide. It does not require physical compression of the breast, can eliminate breast tissue superposition, and provides 3D images at near-isotropic spatial resolution. Earlier generations of breast CT, while demonstrating the concept, were suboptimal due to technological limitations and available components. In the prior funding (R01 CA199044), we focused on the hardware aspects: tested 4 detectors from different vendors, followed by designing, developing, and testing a dedicated breast CT system that employed an offset-detector geometry to enable imaging large breasts. This system achieved high resolution and low system noise and was specifically designed to improve chest-wall coverage and operate at a radiation dose suitable for breast cancer screening. In the pilot study completed under prior funding (R01 CA199044), the system demonstrated visualization of the pectoralis muscle in 177/179 (98.9%) breasts (effective diameter at chest wall: 12.82.1 cm); a remarkable improvement over the 40-78% reported in prior studies, and the mean glandular dose (MGD) was 4.10.9 mGy, which is similar to that reported for digital mammography (4.15 mGy) in the Digital Mammography Imaging Screening Trial (DMIST) study. In this competitive renewal, we focus on image reconstruction followed by a multi-reader, multi-case (MRMC) receiver operating characteristic (ROC) study using prospectively acquired data to evaluate if dedicated breast CT demonstrates improved performance over the current standard, digital breast tomosynthesis (DBT) for breast cancer screening. For image reconstruction, we will investigate a total of 5 techniques that were carefully chosen to encompass the current and emerging methods including the standard Feldkamp-Davis-Kress (FDK) reconstruction, one compressed sensing-based iterative reconstruction that is well-founded on a rigorous mathematical framework, and three deep learning-based reconstruction techniques with increasing sophistication, generalizability, and scientific rigor. The deep learning-based methods include a fully-supervised technique that requires input-reference image pairs and is tuned using physics-based measure, a self-supervised technique that does not require an independent reference image, and a self-supervised technique that is based on both the imaging physics and the mathematics of image reconstruction and does not require independent training data. These methods are not only applicable to dedicated breast CT, but are adaptable to cone-beam CT, in general, and potentially to the commonly used multi-detector CT. The MRMC ROC study is based on well-founded image science and is designed with statistical rigor. To our knowledge, there have been no prior studies comparing the diagnostic accuracy of DBT and breast CT at radiation dose levels suitable for breast cancer screening. This research addresses this unmet need. This research will not only provide pivotal data on the imaging performance of breast CT but also advance the state of image reconstruction that is applicable to the broader field.

NIH Research Projects · FY 2025 · 2016-05

Project Summary/Abstract The postdoctoral training grant entitled “The Neurobiology of Aging and Alzheimer's Disease” is submitted through the University of Arizona on behalf of the Arizona Alzheimer's Consortium. The training faculty chosen from the Consortium consist of highly interactive investigators whose scientific interests include aging, Alzheimer's disease (AD) and AD-related dementias (ADRD) and spans fly, rodent, and nonhuman primate models of aging, as well as human studies in normative and pathological conditions of aging. Six institutions within the Consortium contribute Primary Mentors (Preceptors) and Affiliate Faculty to this Training Program, including: The University of Arizona, Arizona State University, Barrow Neurological Institute, Banner Sun Health Research Institute, The Translational Genomics Research Institute, and Banner Alzheimer's Institute. Faculty have complementary strengths in brain imaging, computer science, genomics, molecular biology, basic systems, behavioral and cognitive neuroscience, experimental therapeutics, computational and statistical analyses of complex datasets and clinical neuropathological approaches. The research is aimed at promoting the scientific understanding of the aging brain, early detection of AD and ADRD and developing effective treatment and prevention therapies. Fellows benefit from a 4 person Professional Development Committee (PDC) whose purpose is to provide guidance on the creation of a collaborative experimental training plan that fosters opportunities to work, learn and produce across institutions. The primary research Mentor, in whose laboratory the Fellow is based, is chosen from the Preceptor list, as is the secondary research Mentor, but they can be from any institution amongst the consortium. The secondary Mentor plays an active role in the research undertaken by the Fellow, which goes beyond making facilities available. A tertiary Mentor is selected on the basis of experimental or pedagogical expertise, and a fourth member of the PDC is chosen from amongst the 6-person leadership team to provide an additional level of oversight. Exposure to the unique Arizona tradition of cooperation and collaboration that knows no institutional boundaries has significantly enriched the skill sets and professional development of our Fellows during the training period. In addition to multi-laboratory research exposure, another unique programmatic aspect takes the form of three workshops per year, varying from 1-3 days, in which all Fellows participate. These include statewide events that Training Faculty attend (Annual Arizona Alzheimer's Consortium Retreat), or that faculty, graduate students and postdocs attend (Alzheimer's Consortium Annual Scientific Conference), as well as broader participation in national and international professional events (e.g., Society for Neuroscience meeting). Our goal to give the Fellow the freedom to draw broadly from facilities and expertise across the Consortium has exposed our Trainees to the power of collaborative, interdisciplinary interactions that can facilitate development of strong experimental designs to test important questions, providing the foundation for productive future work.

NIH Research Projects · FY 2024 · 2015-12

Project Summary Diabetic retinopathy is a disease of both neurons and vasculature in the retina. Visual function deficits and neuronal retinal dysfunction from electroretinograms, are some of the earliest identifiable diabetic retinal problems, especially in the dim light activated rod pathways. Dysfunction of the inner retina - bipolar cells that receive rod input, ganglion cells that receive bipolar cell input and amacrine cells that modulate this pathway- is tied to the development of serious diabetic retinal problems. We have shown deficits in light-evoked responses of rod pathway inner retinal neurons that are not due to cell death, but the neuronal mechanisms that cause these deficits are unknown and the focus of this proposal. We will identify how a loss in dopamine and Ca2+ signaling in the inner retina impacts light evoked vision loss, and we will modulate these pathways to determine if they can be future targets for preventing diabetic retinal dysfunction and the neuronal progression of vision loss. Dopamine is released by dopaminergic amacrine cells to allow retinal adaptation to increasing background light levels and GABA release from other amacrine cells modulates the timing and spatial sensitivity of ganglion cells, increasing the sensitivity of vision to small stimuli. Using a mouse model of STZ induced diabetes we have shown that dopamine receptor sensitivity and light adaptation in one type of ganglion cell and Ca2+ signaling in amacrine cells in the rod pathway are reduced, suggesting that ganglion cell output to the brain is altered leading to diabetic visual deficits. We will use our expertise in analyzing retinal signaling to determine the roles of dopamine and Ca2+ signaling changes in the diabetic retina using physiological signaling, genetically modified mouse models and optogenetic stimulation. In Aim 1 we will determine if the diabetes induced reduction in dopamine levels reduces dopamine receptor sensitivity of the inner retina in the rod pathway, using single cell electrophysiology recordings of the responses of rod bipolar cells and ganglion cells to light and immunohistochemical and in situ hybridization analysis. In Aim 2 we will determine if the release and retinal function of dopamine in light adaptation are reduced in diabetes, using recordings of physiological responses to increasing background light levels at the single cell level, in vitro ERGs and direct dopamine release measurements. In Aim 3 we will determine what diabetes induced changes in amacrine cell Ca2+ signaling cause reduced rod pathway inhibition by stimulating multiple amacrine cell inputs to the rod pathway directly either electrically or optogenetically and blocking specific components of Ca2+ signaling. These proposed studies will significantly expand our understanding of the mechanisms and progression of early diabetic retinal neuronal damage and are part of our long term goal to develop targets for therapeutics to intervene at an early time point of diabetic retinal damage and augment or complement existing treatments to prevent the neuronal progression of vision loss.

NIH Research Projects · FY 2026 · 2015-12

Disregard of genetically-linked biological variation in the design and interpretation of clinical trials of marine omega-3 (n-3) highly unsaturated fatty acid (HUFA) supplementation and specifically limited inclusion of individuals carrying high-activity fatty-acid-desaturase (FADS1) alleles may have masked potential health benefits of individuals at elevated risk for cardiovascular and other chronic diseases. Our work over the past decade has revealed that genetic variation throughout the FADS gene cluster, and the resulting allele-frequency differences among population strata, are significant drivers of the effectiveness of n-3 HUFA supplementation. FADS1 are the rate-limiting steps in the conversion of dietary n-6 and n-3 polyunsaturated fatty acids (PUFA), linoleic acid (LA) and α-linolenic acid (ALA), respectively, to n-6 and n-3 HUFAs, including arachidonic acid (ARA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and their biologically active oxylipin metabolites. Importantly, the FADS1/2 gene cluster variation was recently identified as the largest multimorbidity locus in the human genome. A central discovery from our work and others is the differential synthesis of HUFAs and metabolism to bioactive lipids across genotype-defined strata. Groups enriched for the high-activity FADS1 rs174537-GG allele exhibit efficient conversion of dietary LA to ARA and its pro-inflammatory/pro-thrombotic oxylipins. Evidence in support of this work comes from our genotype subgroup analysis of the VITAL n-3 HUFA supplementation trial affirming that carriers of the high-activity allele, but not lower-activity carriers, randomized to n-3 supplementation demonstrated an ~80 % reduction in myocardial infarction (MI). We then applied two machine-learning methods to matched VITAL cohorts and found strong evidence that n-3 supplementation significantly decreased MI only in high-activity allele carriers (odds ratio 0.178; 95 % CI 0.046–0.69). These data lead to this proposal’s central hypothesis that gene–diet interactions drive a pathogenic imbalance in ARA versus EPA and DHA and their oxylipin products, particularly in high-activity allele carriers. We postulate that n-3 HUFA supplementation will decrease the ratio of ARA to EPA and DHA, providing a major benefit to these individuals. We propose a two-site randomized, double-blind, placebo-controlled, crossover n-3 HUFA supplementation trial that leverages the varied allele-frequency distributions across individuals to test this hypothesis. Specifically, we will supplement participants stratified by FADS genotypes at rs174537 with EPA-enriched n-3 HUFAs and evaluate changes in the balance of HUFA-containing lipids and oxylipin products, as well as inflammatory and clinical biomarkers linked to chronic disease. This trial will provide critical mechanistic evidence for genotypelinked biological heterogeneity in differential biological response to n-3 HUFAs. Such mechanistic data are essential to the future design of definitive precision nutrition clinical trials to prevent chronic diseases and reduce residual cardiometabolic risk.

NIH Research Projects · FY 2026 · 2015-07

Project Summary Project Summary: Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases worldwide. The pathogenic mechanism remains incompletely understood. Bile acids (BAs) are endocrine hormones that are important for the biological processes. Dysregulation of BA metabolism may cause cholestasis, liver cancer, NAFLD, etc. Activation of BA receptors FXR and TGR5 has been shown to protect against NAFLD in mice and clinical trials. Transmembrane protein 141 (TMEM141) belongs to the TMEM protein family that consists of proteins of mostly unknown functions. So far, nothing is known about the biological functions of TMEM141. In this project, we plan to investigate the role of hepatic TMEM141 in the pathogenesis of NAFLD. We will investigate whether and how hepatic TMEM141 regulates the development of NAFLD and bile acid metabolism, and how hepatic TMEM141 expression is regulated in NAFLD. We will use a number of genetically modified mice and a complementary approach to accomplish our goals.

NIH Research Projects · FY 2026 · 2014-09

The nation, including the state of Arizona with a population that is 38.5% Latino and Native American, suffers from tremendous health disparities contributed to, in part, by the absence of a diverse biomedical research and healthcare workforce. Never has there been a greater need for health equity that needs to be addressed by a diverse research workforce and infrastructure than during and following the Coronavirus Disease (COVID-19) pandemic during which time our minority communities were disproportionately affected by the pandemic due to respiratory failure and pulmonary-sleep manifestations of post-acute sequelae of COVID (PASC). In this renewal application, we propose to build upon our past successes with innovative approaches and best practices to advance research education tailored for qualified PRIDE trainees. The overarching goal of our program is to provide advanced research training experiences and long-term mentoring in an interprofessional environment to qualified candidates -- junior faculty and transitioning postdoctoral scientists from diverse backgrounds, including those from groups that are underrepresented in the biomedical sciences -- who are committed to investigating the factors responsible for differences in health among populations as they pertain to lung, sleep-related breathing disorders, and the consequences of the pandemic to these systems. Our proposal is to redesign, organize, and implement a NHLBI mission-focused Summer Institute program that will support AiRE and program faculty to nationally recruit eligible and highly-qualified individuals in order to provide advanced training experiences and long-term mentoring that will enable them to develop a research program and work with their home institution to obtain NIH funding and develop their career and gain leadership skills. The research training experiences will be tailored to the trainee and designed to enhance their research skills, experiences, and knowledge base in lung and sleep-related breathing disorders research with attention to the specific scientific area of infectious and immunobiological consequences of the pandemic on lung and sleep using cross-cutting methodological approaches. The broader goal of the AiRE training program is to create a rigorous interprofessional research training program that attracts highly-qualified early career faculty and transitioning post-doctoral scientists, offering them the academic and collaborative research experience that supports a successful and productive career in the study of disparities in lung, sleep-related breathing disorders, and their pandemic-related consequences. The impact would be to engender a diverse biomedical research workforce in lung, sleep, and their pandemic-related consequences that can help us better understand health disparities and promote health equity.

NIH Research Projects · FY 2025 · 2014-07

PROJECT SUMMARY Hypertension is the No. 1 identifiable risk factor for disease burden and deaths worldwide. Endothelial dysfunction contributes to the development of hypertension. Once hypertension develops, kidney injury ensues in susceptible patients. Albuminuria and proteinuria are often modest in hypertension but are prominent in subgroups such as African American hypertensive patients and are predictors of unfavorable prognosis. microRNAs regulate target protein expression primarily by inducing mRNA degradation or translational repression. Like many regulatory mechanisms such as signaling pathways and transcriptional factors, microRNAs are potentially promiscuous but may achieve regulatory specificity in a defined cellular context, a notion that has been reinforced and validated with recent technological advancement. Several microRNAs have emerged as powerful regulators of cardiovascular and renal functions with strong relevance to human disease. miR-204 is one such “high value” microRNA. The most striking and intriguing finding of our study of miR-204 was an apparent dissociation of miR-204’s effects on hypertension and renal injury in mice treated with uninephrectomy, angiotensin II, and a high-salt diet. The Unx/AngII/salt model typically exhibits a sustained increase of mean arterial pressure of ~40 mmHg and develops renal injury including albuminuria. Global miR-204 gene KO (Mir204-/-) attenuated the development of hypertension in this model, reducing mean arterial pressure by up to 30 mmHg compared to wild-type mice. In contrast, renal injury in the Unx/AngII/salt model was exacerbated in Mir204-/- mice, showing several fold greater albuminuria. These striking findings suggest that the functional role of miR-204 might be broad and highly tissue-specific. We have generated seven (7) new strains of mice to examine tissue-specific roles of miR-204 and the underlying mechanisms. Based on strong preliminary data, we hypothesize that miR-204 in endothelial cells permits the full development of hypertension (Aim 1) while miR-204 in podocytes protects against the development of albuminuria in hypertension (Aim 2). In both aims, we will examine the pathophysiological role of miR-204 in specific cell types, investigate the role of specific target genes that have been nominated by RNA-seq and other analyses, and study cell type-specific regulation of mRNA translational activities. The proposed study will drive a new dimension of hypertension and tissue injury research where a gene or pathway acts in one cell type to promote hypertension and another cell type to attenuate renal injury. The study will elucidate novel and critically important aspects of the complexity of these diseases, promote tissue-targeted therapeutics, and advance the recently reinforced notion of cell type specificity of microRNA function.

NIH Research Projects · FY 2025 · 2014-01

Summary Our long-term goal is to understand diaphragm contractility in health and disease, and the role of titin therein. The diaphragm muscle drives respiration and is constantly subjected to mechanical loading. Changes in mechanical loading rapidly affect diaphragm contractility, e.g., within hours of diaphragm unloading during mechanical ventilation (MV) in ICU patients, severe diaphragm weakness ensues, contributing to difficult ventilator weaning. The pathophysiology of diaphragm weakness is incompletely understood. In the proposal at hand, we will study the role of depressed interactions between myosin and actin, the two key contractile proteins in myofibers, and the role of titin in force depression. Our pilot data suggest that unloading of the diaphragm causes the myosin motors to adopt the so-called ‘super-relaxed state’ (SRX). Once in the SRX state, less myosin motors are available for binding to actin, and less force can be generated. We will also study the role of the giant protein titin in releasing myosin from the SRX state and investigate whether during MV-induced unloading of the diaphragm, this mechanism is perturbed. Aim 1 will determine the SRX state of myosin in the diaphragm of ventilated ICU patients. We will use our diaphragm biopsies of ICU patients to study the contractile force of diaphragm myofibers and determine the number of myosin motors that attach to actin during activation. We will combine mechanics with X-ray diffraction and biochemical assays to resolve with nanometer resolution the position of myosin motors relative to actin during activation and the proportion of myosin in the SRX state. In Aim 2 we will determine the role of posttranslational modifications in the SRX of myosin. Phosphorylation of regulatory light chains (RLC) regulates the SRX state of myosin and we will apply mass-spectrometry on the diaphragm biopsies to determine the phospho-proteome, and we will establish whether there is a cause-and-effect relation between RLC phosphorylation and SRX by performing RLC exchange experiments into patients’ myofibers. To critically test whether changes in RLC are caused by diaphragm inactivity, we will ventilated healthy rats, and study RLC phosphorylation. Aim 3 will determine whether, in addition to RLC phosphorylation, direct mechanical effects of titin on myosin induce SRX. Titin-based passive tension strains the myosin filament which regulates the SRX state. We will study whether the reduction in titin-based passive tension, due to diaphragm unloading during MV, reduces the strain in the myosin filament and increases SRX. We will use mouse models with genetically engineered increased or decreased titin-based passive tension and study the effect of MV on the SRX state of myosin. The innovation of this proposal lies in the novel research foci and guiding hypotheses, unique diaphragm biopsies from patients, and its novel tools and mouse models, The proposal’s integrative approach is expected to lead to a major step forward in our understanding of diaphragm function and titin’s role therein.

NIH Research Projects · FY 2026 · 2013-08

Naphthalene (NA) is a ubiquitous and highly abundant pollutant found in vehicle exhaust, fossil fuels and wildfire smoke. NA causes nasal and lung toxicity, including tumors, in rats and mice, respectively, and is a possible human carcinogen. The mechanism of NA carcinogenicity, which may involve both genotoxic and non- genotoxic events, is not clear and may involve several reactive NA metabolites. A prerequisite for NA toxicity is bioactivation by cytochrome P450 (CYP) to form NA-epoxide (NAO) which can undergo further metabolism in the lung and liver to 1,2-naphthoquinone (1,2-NQ). Both NAO and 1,2-NQ can react with DNA. We have recently uncovered a significant contribution of liver-generated NA metabolites to acute lung toxicity in vivo; identified several NQ-DNA adducts in the mouse lung, liver, and blood following NA exposure and in blood samples from human firefighters and lung biopsy samples of lung cancer patients; and gained novel insights on the toxicokinetics of various NA metabolites. These exciting findings led to the current proposal, to test the novel hypothesis that liver-generated reactive NA metabolites are transported to the lung where they contribute to NA's cytotoxicity, genotoxicity, and carcinogenesis. We are well positioned to address this hypothesis due to the genetically modified mouse models we have developed, the recent methodologic advances we have made in metabolite and DNA adduct characterization, and the exposure systems we have in place to expose mice acutely and/or chronically to NA vapor at defined doses. A series of mechanistic and translational studies will be carried out in four Specific Aims that will 1) identify key metabolic events that control NA DNA adduct formation in the lung, 2) determine whether serum albumin facilitates transport of liver-generated reactive NA metabolites to the lung, 3) identify whether key metabolic events in the liver mediate NA-induced lung tumorigenesis in vivo, and 4) characterize profiles of NA metabolites and DNA adducts in blood and urine from firefighters. Our long-term goal is to identify metabolic mechanisms that benefit risk assessment for chemical-induced lung toxicity and carcinogenesis in humans. The proposed studies remain focused on metabolic mechanisms underlying NA respiratory toxicity and address mechanisms for the hepatic contribution to lung toxicity, link hepatic NA metabolism to lung tumorigenesis, and examine biomarkers of NA's genotoxicity in humans. These studies define key metabolic events that influence NA-mediated lung toxicity and carcinogenesis. Success of the proposed studies will move the field of mechanistic toxicology forward toward a better understanding of fate and importance of circulating reactive metabolites and shift the paradigm for NA risk assessment, by providing proof- of-principle evidence for the importance of circulating (as well as locally generated) NA metabolites in NA's genotoxicity and lung tumorigenesis, identifying accessible biomarkers for monitoring potential NA genotoxicity in humans, and advancing the understanding of hepatic NA bioactivation as a risk factor that may predispose individuals to NA-induced lung carcinogenesis.

NIH Research Projects · FY 2026 · 2012-09

ABSTRACT The duration of antibody-mediated immunity varies widely with the specific immunization or infection for reasons that remain difficult to understand or predict. The specific features of molecular circuitry in B cells and plasma cells that are responsible for these differences have still not been defined. In this application, which requests a second renewal of our R01 award, we seek to objectively define 1) molecular programs and cellular trajectories that lead to long-lived plasma cells in primary responses; 2) transcriptional and epigenetic features of memory B cells that precede durable recall responses. We will employ two BTB-POZ transcription factors that functionally oppose each other to gain molecular footholds on programs that control plasma cell lifespan. Modern single cell transcriptional and epigenetic programs will be employed alongside classic model antigens and adjuvants and new genetic tools to manipulate genes in vivo. Though the experimental plan is fundamental and basic in nature, the hope is that this work will lead to a set of rules that help predict and optimize the duration of humoral immunity.

NIH Research Projects · FY 2026 · 2012-07

Project Abstract Contemporary models of spatial navigation and episodic memory (memory for events) postulate that their underlying computations emerge primarily from shared neural mechanisms within the medial temporal lobes. As part of the last two rounds of funding for this competitive renewal, we have begun to delineate important cognitive and neural differences between navigation and episodic memory. Our emerging new framework argues for navigation as a sensory-driven cognitive motor skill involving extracting spatial regularities and episodic memory as primarily internally driven and involving ordinal placeholders. We hypothesize that navigation and episodic memory therefore involve partially distinct brain regions and macroscale networks, although where and how these differences emerge in the brain remains an area of active exploration. Here, we test novel aspects of this theoretical framework: how pre-existing knowledge differentially affects the acquisition of new episodic memories compared to navigation-related representations over both longer (days and weeks; Aim 1) and shorter (hours; Aim 2) intervals. Throughout, we propose meaningful alternative models, including the idea that connectivity to the hippocampus and neocortex, and cortical macroscale networks outside of the hippocampus, play critical and unique roles in episodic memory compared to navigation. In Aim 1, using high-resolution fMRI, we propose to employ three different experiments to compare how schema (pre-existing spatial knowledge) and scripts (pre-existing temporal knowledge) differentially interact with new learning in the context of episodic memory and navigation. Together, the outcomes from these experiments will provide mechanistic insight into how we organize episodic memories and navigation-relevant knowledge over longer intervals that could be meaningful for cognitive rehabilitation. In Aim 2, we focus on how episodic memory interacts with navigation and pre-existing knowledge over shorter-term intervals (hours) by studying mental simulation before and after navigation. Mental simulation involves actively remembering or planning experiences and has direct links with cognitive processes central to episodic memory, particularly in our three different proposed experiments. Here, we will employ time-resolved intracranial EEG in conjunction with Dr. Brad Lega at University of Texas Southwestern to better identify the mnemonic content of both navigation and mental simulation, including a causal manipulation involving the muscarinic acetylcholine antagonist scopolamine and single cell recordings. Together, the experiments in Aim 2 will provide novel insight into the mechanistic basis of episodic memory and navigation-related representations. Such mechanistic could be helpful in developing neurostimulation or pharmacological protocols (e.g., involving acetylcholine) that could be used to bolster either impaired memory or navigation function following stroke, seizure damage, or other brain injuries affecting hippocampal function.

NIH Research Projects · FY 2026 · 2011-08

University of Arizona's High School Student NeuroResearch (NR) Program (HSNRP) nurtures, trains, and sustains the spirit of inquiry in a growing diverse, connected workforce pipeline and faculty peer/near peer support network. Over the next 5 years, HSNRP will offer annually 10 talented, motivated Arizonawide high school (8 weeks) and 3 progressing undergraduate students (10 weeks) full-time closely mentored hands on-brain on basic, translational, clinical and popula- tion research experiences emphasizing the workings and disorders of the normal and abnormal brain (“favorite organ” of interviewees), spinal cord, and peripheral nervous system and encourage continuing involvement in more advanced research leading to science/medical/health-related careers. HSNRP leverages the strong infrastructure, effective recruit- ment/retention strategies, engaging student/faculty/near peer relationships, and outstanding trainee productivity of our long-standing federally funded multidisciplinary disadvantaged high school/undergraduate/medical student summer research programs and year-round enrichment. Interacting together, HSNRP trainees will be integrated into an innovative, internationally recognized question-based Summer Institute on Medical Ignorance (SIMI) with novel mobile-accessible software platforms designed for collaboration and interweaving biomedical Knowns and Unknowns – what we know we don't know (research), don't know we don't know (discovery), and think we know but don't (error), i.e., unanswered/ unasked questions and unquestioned answers. Bringing together multilevel trainees, the summer curriculum features informal triweekly general and NR biomedical topical seminars and faculty/SIMI alumni "life stories" with periodic enrichment activities year-round emphasizing "translating translation and scientific questioning," and introducing the language and principles of pathobiology, neuro-anatomy/physiology/pharmacology; clinical correlations, laboratory/ leadership/multimedia communication skill practice, social networking, and sustained career advising. Within multiple basic and clinical departments and specialized Centers of Excellence and overseen by an energetic experienced multi- disciplinary HSNRP leadership team, research encompasses cross-cutting themes and in vivo, in vitro, in situ, in silico, and modeling approaches to neurobiology/disorders ranging from Parkinson and Alzheimer disease, epilepsy, traumatic brain injury, hydrocephalus, muscular dystrophies, headache, pain/addiction, to sleep disturbances, brain tumors, deep brain stimulation, molecular psychiatry, cognition, blood-brain barrier/neuroprotection, neuroimaging, neurogenomics/ proteomics, neuroengineering, cerebral hemo/lymphovascular dynamics, stroke, and health disparities. Based on our ~35-year track record and access to large diverse pools of disadvantaged/URM Arizona students reflected in 787 SIMI- trained high school students (including110 HSNRP alums) followed to date with substantial numbers in or working toward science and specifically NINDS mission-related basic science/clinical/teaching careers, we expect HSNRP to continue to extend and enlarge the NR diversity pipeline and improve neurohealth literacy through community engage- ment. Ongoing evaluation includes feedback surveys, database registry, curiosity scales, short-/long-term followup, and individual portfolios to document efficacy and the training model's sustainability and promote diversity and networking.

NIH Research Projects · FY 2025 · 2009-09

American Indians/Alaska Natives (AIAN) are disproportionately impacted by the burden of cancer. Compared to other racial and ethnic groups in the United States (US), AIAN individuals face higher rates of many cancers, later stages at diagnosis, worse outcomes after diagnosis, and lower rates of cancer survival. The Partnership for Native American Cancer Prevention (NACP), a 21-year collaboration between Northern Arizona University (NAU) and the University of Arizona Cancer Center (UACC), has made impactful strides that successfully address the causal factors that drive AIAN cancer inequities. NACP has had a significant positive impact on the pipeline of AIAN individuals seeking careers related to cancer health research, including the training of Native investigators poised to be leaders in this field. NACP has been a driver of institutional change at both NAU and UACC by fostering an increase in cancer research capacity at NAU and health disparity-focused research at UACC and by elevating both institutions’ commitments to serving AIAN students and communities. NACP has built a strong foundation of relationships with tribal communities, governments, and other partners, based on trust and respect, and this is resulting in an acceleration of the positive impacts driven by NACP’s activities, both present and future. NACP is poised to realize past investments while sustaining current relationships and expanding interactions to additional tribal communities in Arizona. The NACP remains committed to its core goals of reducing the burden of cancer within AIAN populations through research and community engagement, growing the number of AIAN investigators participating in the cancer research workforce, and increasing the total number of investigators focused on cancer health disparities within Native Arizona communities. While these overall goals remain consistent, NACP is introducing an operational framework to systematically incorporate Indigenous perspectives as a core reference to guide and thread together its work. NACP will embrace the ‘two-eyed seeing’ paradigm, which seeks to “see from one eye with the strengths of Indigenous knowledges and ways of knowing, and from the other eye with the strengths of Western knowledges and ways of knowing, and to use both of these eyes together for the benefit of all.” Aim 1. To engage in bidirectional communication grounded in the principle of reciprocity with our AIAN community partners to promote best practices with respect to cancer health and to develop research priorities and programs that address AI cancer health disparities. Aim 2. To grow the pipeline of cancer-focused Indigenous researchers and health care professionals through educational and training programs tailored to high school, undergraduates, graduate students, junior investigators, and early-stage investigators. Aim 3. Conduct impactful cancer disparity focused research that is informed by and inclusive of tribal community priorities and concerns.

NIH Research Projects · FY 2025 · 2007-04

ABSTRACT Our laboratory was the first to demonstrate unequivocally that several isoforms of small conductance Ca2+- activated K+ channels (SK or KCa2 channels) underlie Ca2+-activated K+ current (IK,Ca) in cardiomyocytes. Since our original reports, knowledge of cardiac SK channels in the field has been greatly expanded. Studies by our group and more recently by others, have provided evidence to substantiate the important roles of SK channels in the heart. Interests in cardiac SK channels are further fueled by recent studies suggesting the possible roles of SK channels in human arrhythmias and atrial fibrillation (AF). Therefore, SK channel may represent a novel therapeutic target for atrial arrhythmias. Moreover, SK channels are upregulated in heart failure (HF). To our knowledge, they are the only K+ channels that are upregulated in HF, underpinning the importance of this class of channels in normal and diseased hearts. Significant gaps in our knowledge and seemingly contradictory findings on SK channel function in cardiac disease mechanisms are our motivations for the next grant cycle. These are the challenges: 1) Blockade of SK channels has been shown to be both anti-arrhythmic and proarrhythmic in various models; and 2) SK channels are upregulated in HF. However, the mechanisms for the observed upregulation remain incompletely understood. Thus, this multidisciplinary proposal will combine experimental and computational studies, taking advantage of complementary expertise from four different laboratories in functional studies, optogenetic tools, and computational modeling to successively address the multifaceted SK channel remodeling in diseased hearts. SK current is enhanced during -adrenergic (-AR) stimulation, especially in female animals, therefore, the sex-specific roles of SK channels will be tested. The proposed study represents the necessary and critical steps to disentangle the highly complex and sex-specific SK channel remodeling in HF, the unique K+ channel that helps to compensate for the much-needed “repolarization reserve” in HF.

NIH Research Projects · FY 2025 · 2006-08

PROJECT SUMMARY – OVERALL Each year ~1.5 million American women enter into the perimenopause, a midlife neuroendocrine transition state unique to the female. As of 2020, there are 45 million US women over the age of 55. Globally, there are currently over 850 million women aged 40-60 years of age. Worldwide women have a 2-fold greater risk for developing Alzheimer’s. The mission of the Perimenopause in Brain Aging and Alzheimer’s Disease Program Project is to discover biological transformations in brain that occur during the perimenopausal transition that lead to endophenotypes predictive of risk for Alzheimer’s disease (AD). Research goals are to identify the mechanisms by which these transformations occur and to translate these discoveries into strategies to prevent or delay conversion to AD. Our research has shown that the greater risk for AD is not because women live longer than men but because the disease can start earlier in women, at midlife during the perimenopausal transition. In the Perimenopause in Brain Aging and Alzheimer’s Disease program of research, we advance mechanistic, clinical and population discovery science and translate these discoveries into a platform for precision medicine to prevent, delay and treat Alzheimer’s disease. Herein we specifically focus on the complex interaction between APOE genotype and the metabolic and immune systems that initiate and drive pathologies of Alzheimer’s. To achieve our mission, we have developed a focused research center model with an integrated set of four Projects and three Cores. Projects 1, 2 and 3 are basic, mechanistic and preclinical translational science investigations of the perimenopausal brain utilizing humanized APOE mouse models relevant to Alzheimer’s risk and to human perimenopause. These projects investigate the molecular, cellular and systems biology of immune signaling in brain and periphery that initiate and drive development of Alzheimer’s disease in brain and autoimmunity in peripheral organs. Project 4 investigates development of the endophenotype of early stage Alzheimer’s disease in perimenopausal to postmenopausal women using multi-modal brain imaging and analyses of peripheral biomarkers. All Projects and Cores are highly integrated and supported by a suite of enabling strategies and technologies. Outcomes of proposed aims will generate a mechanistic foundation on which to conduct hypothesis driven medical informatics, develop neuro-immune biomarkers specific to stages of brain aging and a platform for precision neuro-immune therapeutics. The Perimenopause in Brain Aging and Alzheimer’s Disease program of research addresses key strategic goals of the National Institutes on Aging’s 2016: Aging Well in the 21st Century: Strategic Directions for Research on Aging, specifically Goals A (1,2,3,7,8,11) & D (1,2,4).

NIH Research Projects · FY 2025 · 2005-09

PROJECT SUMMARY / ABSTRACT Diabetes affects nearly 10% of the adult population (30 million), and these numbers are expected to double by the year 2050. The pathophysiology of diabetes profoundly impairs all tissue reparative processes, leading to chronic non-healing wounds in affected patients. Diabetic foot ulcers affect between 15-30% of all diabetic individuals and represent the leading cause of lower limb amputations in the United States. Conventional methods to treat diabetes, such as with insulin or oral hypoglycemic agents, can control the disease but do not prevent diabetic complications, as demonstrated by continued progressive organ dysfunction even decades after medical optimization. This highlights a clear need for new therapeutic approaches. Over the past 15 years of NIH funding, our laboratory has made important contributions to our understanding of the critical molecular and cellular pathways in normal and diabetic tissue repair. We have identified hyperglycemia-related impairments in both the local microenvironment and progenitor cell homing and cytokine production that contribute to the pathogenesis of diabetic complications. We have demonstrated that diabetes results in depletion of critical cell subpopulations, resulting in decreased neovascularization and impaired tissue healing. To understand the effects of diabetes on cell population dynamics with greater precision, we have developed novel single cell “-omics” techniques to identify critical perturbations in cell subpopulations at the single cell level. It is our fundamental hypothesis that diabetes alters the “cellular ecology” of heterogeneous cell populations involved in tissue repair and that normalization of those cell subpopulations can treat or reverse diabetic complications. In this proposal, we will integrate emerging multimodal -omics technologies to definitively characterize the behavior of cell subpopulations in diabetic complications, including wound healing. We will extend this work therapeutically by using cell-based approaches to normalize these defects to treat and prevent diabetic complications. To begin, we will employ a novel multiplex approach for high-throughput single cell sequencing to definitively characterize the behavior of resident tissue and progenitor cell subpopulations in human diabetic and non-diabetic wounds (Specific Aim 1). We will then confirm these human observations in animal models and define these changes with spatial resolution by integrating single cell sequencing with next-generation spatial transcriptomic and proteomic technologies and precisely delineate where in the three-dimensional wound environment these differences exist (Specific Aim 2). Finally, we will use this information to optimize the systemic delivery of cell-based therapeutics in order to prevent or reverse diabetes-induced defects in relevant cell populations and thereby correct diabetic complications (Specific Aim 3). Taken together, this novel approach for identifying, spatially characterizing, and correcting subpopulation deficits in diabetic complications will provide new insights into diabetic pathophysiology and inform novel strategies to prevent and treat these complications.

NIH Research Projects · FY 2025 · 2005-09

TITLE: Role of Cardiac Myosin Binding Protein-C in the Regulation of Myocardial Contraction ABSTRACT: The overall goal of this project is to understand how cardiac myosin binding protein-C (cMyBP-C) regulates heart muscle contraction and relaxation. cMyBP-C is a critical regulator of cardiac function, with dysfunction of cMyBP-C commonly occurring in heart failure and mutations in cMyBP-C being the most common genetic cause of HCM. However, the regulatory effects of cMyBP-C are complex and most likely involve dynamic interactions with both thick (myosin containing) and thin (actin containing) filaments, making it challenging to sort out what are likely to be reciprocal or interrelated effects of cMyBP-C on each filament. To overcome these and other challenges, we recently developed a novel method that allows us to rapidly remove and replace (“cut and paste”) cMyBP-C at its endogenous position in sarcomeres, thus affording us the opportunity to quickly test cMyBP-C effects on both filament systems. In Aim 1 we will test the functional significance of the “middle domains” of cMyBP-C which were recently implicated as regulators of thick filament relaxation by stabilizing the “interacting heads motif” conformation of myosin on the thick filament as well as novel regulators of cMyBP-C function including Ca2+-Calmodulin (CaM) that we show in exciting new preliminary data may regulate cMyBP-C binding to the thin filament. Because HCM mutations and posttranslational modifications (PTMs) also occur in the middle domains and have been correlated with diastolic dysfunction we will also test effects of selected mutations and PTMs in these domains. Experiments will include measurements of steady state force in permeabilized cardiomyocytes as well as rates of activation and relaxation in myofibrils and the activation state of thick filaments determined by myosin DRX/SRX ratios. In Aim 2 we will use the cut and paste approach combined with X-ray diffraction to determine structural effects of cMyBP-C on the activation/relaxation states of thick and thin filaments in sarcomeres. Specifically, we will test the hypothesis that C-links between thick and thin filaments modulate the on/off states of thick and thin filaments and contribute to thick filament activation in response to passive stretch or active load. Results from these studies will provide new insights into how cMyBP- C regulates heart function during health and disease.

NIH Research Projects · FY 2025 · 2003-12

Project Summary: The cardiac thin filament is the essential regulator of cardiac contractility and relaxation at the molecular level. It is comprised of five discrete proteins: cTnC, cTnI, cTnT, actin and tropomyosin that have co-evolved to sustain efficient cardiac performance at rest, during exercise and, importantly, to respond to pathologic stressors. Mutations in genes encoding each of these proteins have been definitively linked to the development of a range of human genetic cardiomyopathies, including hypertrophic (HCM) and dilated (DCM) forms. Despite 25 years of study by many groups including ours, to define the direct link(s) between the biophysical insult and the resultant complex cardiomyopathy, many questions remain and significantly limit our ability to use genotype to prognosticate and eventually even treat individuals with genetic cardiomyopathies. The recent development of Mavacamten, a first-in-class, targeted myosin inhibitor is a game-changing advance that was predicated on decades of basic research into the fundamental biology of the sarcomere. Thus, the question is no longer “if” we can target the sarcomere, but for the thin filament the question is “what function to target” and eventually “when to treat”. The cardiac thin filament is a highly dynamic allosteric “machine” where most of the component proteins are comprised of a-helices connected by variably sized unstructured linkers, where dynamic flexibility is the rule, not the exception and this has limited the availability of high resolution structure for these regions. Most of the known pathogenic mutations in cTnI and cTnT are clustered within these highly flexible domains, where there is likely a “distribution” of tolerance, whereby mutations impair function (enough to cause disease) but do not break it. We thus propose that by examining the range of these dynamic perturbations within these domains we can identify new structural and dynamic disease mechanisms that can be functionally binned, studied and modulated. We provide proof-of-principle preliminary data in this proposal. Over the recent funding period we expanded our structural methodologies to include Time-Resolved FRET with a Single Donor – Dual Acceptor approach that allows us to use actin as an anchor to refine highly flexible structures. We will next use known, highly divergent (HCM vs DCM) mutations within each flexible domain to probe both structure and dynamics with the premise that these mutations will define the limits of “tolerability” in either direction and use spectroscopy and measurements of Ca2+ dissociation and association kinetics coupled to computation to define and test these hypotheses. Finally, we will “close the loop” by utilizing our existing extensively characterized transgenic animal models based on the same mutations used to set our limits and perform 3-timepoint RNA-Seq to discover unique early transcriptional signatures to help link these perturbations to the resultant early remodeling cascade that eventually leads to distinct patterns of ventricular remodeling. The long term goal is to use this coupled structural – dynamic – transcriptomic platform to identify new targets, both primary and secondary for future therapeutics and even biomarker discovery.

NIH Research Projects · FY 2026 · 2001-08

The premise of this project is that studying immune aging in laboratory mice that are experiencing a natural burden of major infectious, psychological and physical stressors will better mirror and inform immune aging in humans. Old age is accompanied by increased vulnerability to infectious diseases, due to the aging of the immune system. Immune aging is, in turn, substantially influenced by the presence of a lifelong, persistent infection with the cytomegalovirus (CMV). In the past period of this award, we studied mouse CMV (mCMV) in isolation (as a latent persistent mono-insult). We concluded that mCMV substantially degraded the healthspan of mice, but only in the presence of major stressors, such as ionizing radiation or another infection. We propose to advance studies of mCMV and immune aging in mice experiencing a natural burden of major infectious, psychological and physical stressors that are likely to be encountered by humans repeatedly during the lifespan. We hypothesize that life-long latent mCMV infection contributes to the demise of T-cell and global immune function, directly proportional to the stress-induced viral reactivation and loss of immune control over mCMV. To test this, animals will be exposed to (i) low dose ionizing radiation; or (ii) low dose stress hormone corticosterone; (iii) non- specific pathogen free (non-SPF) microbiota, which has been shown to induce the immune system in SPF mice to become similar to that of humans. We will test how this impacts mCMV reactivation and immune responses to vaccination and lethal infection. We will ask: SA1. Do repeated individual stressors, commonly experienced by humans, worsen the impact of mCMV on immune function with age? SA2. Is the impact of life stressors on immune aging and overall health in mCMV-positive mice caused by CMV DNA replication/reactivation? These experiments will, for the first time, introduce key physiological variables of importance to human immune aging into a well-controlled murine model of mCMV, aging and infection. We anticipate that this poly-insult model will substantially mirror immune aging in humans, and provide ground for new discoveries that will pave the way for combined antiviral and stress-control therapies to improve T cell and overall immunity and healthspan outcomes in aging.

NIH Research Projects · FY 2025 · 1997-07

OVERALL: ABSTRACT The University of Arizona Cancer Center (UACC) is the only National Cancer Institute (NCI)-designated Cancer Center headquartered in the state of Arizona. UACC was established over 45 years ago and has served as the driver of cancer research at the University of Arizona, within the Catchment Area, and in the state of Arizona. UACC’s vision is to be a national leader for overcoming cancer risks, improving treatments, training talented scientists and providers, and engaging communities though a shared determination to discover, innovate, and address health needs of their population. This vision is realized through UACC’s mission, which is to alleviate the burden of cancer on patients and families within a five-county Catchment Area and beyond through: (1) a quest for scientific discovery through interdisciplinary collaboration and team science; (2) increasing access to the latest approaches throughout a patient’s cancer journey; (3) preparing generations of researchers and health professionals to fight each day against cancer and the challenges affecting underserved populations; and (4) providing national leadership in engaging populations with respect, consistency, and shared conviction. Joann Sweasy, PhD, was appointed UACC Director in 2020 and has promoted engagement of cancer center leaders and members around this mission and vision. Sweasy initiated a robust strategic planning process that led to development of the 2020–2025 Strategic Plan, “Bear Down on Cancer”, emphasizing transdisciplinary collaboration around three Center-wide scientific themes that are highly relevant to UACC’s Catchment Area: Cancer and the Environment; Cancer Imaging and Bioengineering; and Novel Therapeutics and Preventive Interventions. UACC draws its 151 members from 11 colleges and 36 departments. Total funding is $33.8M (DC), which includes $26.1M in peer-reviewed funding of which $11.6M (44%) is from NCI and $13M (50%) from other National Institutes of Health agencies. This represents a nearly 20% increase in peer-reviewed funding compared to the prior project period. Of note, UACC has received $2.7M in NCI supplements during this period. In 2020, 2,835 patients were accrued to interventional trials, a more than 300% or 3-fold increase in accruals compared to 2016. During the project period, the Office of Community Outreach & Engagement was launched, and a population health assessment was initiated to profile the Catchment Area more deeply; an Office of Cancer Research Training & Education Coordination was also established to integrate and expand cancer pipeline programs and junior investigator mentorship. Under Sweasy’s direction, Research Programs and Shared Resources were also reviewed and reorganized for greater impact; internal funding opportunities and Innovative Working Groups were expanded to promote transdisciplinary collaboration and coordination; a multi-year recruitment effort was initiated based on the new Strategic Plan; rigorous methods were implemented to ensure cancer relevance of metrics; and an unprecedented collaboration was formed with Banner Health – Tucson to foster clinical research and care.

NIH Research Projects · FY 2026 · 1997-04

The Strategic Vision of the Southwest Environmental Health Sciences Center (SWEHSC) is to facilitate and implement innovative research and community engagement aimed at understanding the mechanisms underlying environmental health (EHS) risks and disease among people living in arid environments. Importantly, the SWEHSC strategic vision supports our long-term goal of uniting interdisciplinary scientists to study environmental effects on the health and well-being of people in the arid Southwest environment. Because the unique conditions of the arid desert Southwest environment mirror those of many other global desert climates, the interdisciplinary research conducted by the SWEHSC could also improve the lives of the 2.1 billion people globally who live in arid lands. As extreme weather increases the burden on human health through water and respiratory exposures due to drought, wildfires, and decreasing water supply, the arid Southwest serves as the proverbial ‘canary in the coal mine’ for the resulting health effects. Specifically, research within the SWEHSC focuses on 1) routes of exposure in arid environments, including exposure to groundwater contaminants and inhalation, 2) the adverse health outcomes following inhalation of air pollutants, and 3) the molecular pathways of adaptive responses to environmental exposures such as arsenic and ultraviolet light. EHS factors that are prevalent in the desert Southwest will help forecast the concerns related to other arid lands. Accordingly, our mission impacts not only the health and well-being of people in the arid Southwest, but also the billions of people across the planet affected by arid environments. The SWEHSC will continue to improve the health of people in arid lands by developing rational approaches to identifying and mitigating hazardous environmental exposures. The geographic location of the SWEHSC provides unique research opportunities to identify and address basic environmental health hazards that impact these populations. Moreover, strengthening ties between SWEHSC faculty and academic and governmental agencies in Mexico enables impactful binational EHS initiatives, with the tangible ability to improve public health along the US-Mexico border and for broad translation to global stakeholders.

NIH Research Projects · FY 2026 · 1997-04

for Overall Center The University of Arizona Superfund Research Center addresses the unique human health risks encountered in the U.S. Southwest, a region with a rich history of metal mining and generation of mine wastes. We (and others) have demonstrated how mining communities in this region experience layers of exposure to hazardous metals and metal(loid)s. These layers of exposure include drinking water, but uniquely in this environment, substantial exposure occurs from inhalation and ingestion of metal(loid)-contaminated mine waste dusts transported into homes and exterior environments including soils, gardens, and play areas. An added layer is that these populations are simultaneously subjected to extensive, continuous inhalation exposure to fungal spores. Little information exists on the risks associated with inhalation exposure to metal(loid)-contaminated dusts, let alone risks from co-exposure to metals and fungal spores. Yet the outcomes of inflammation-related lung injury following inhalation exposures are serious, ranging from asthma to fibrosis, chronic obstructive pulmonary disease, and cancer. Our overall goal is to construct a mechanistic model of how chronic exposure to mining-impacted dust that is co-contaminated with metal(loid)s and fungal spores contributes to the development of nonmalignant lung diseases, then implement this model to predict exposures and associated health outcomes, to inform public health prevention in communities neighboring mine waste sites and design remediation-based interventions to exposure. To achieve this goal, we have four highly integrated research projects and four cores that will work to measurably reduce non-malignant lung diseases in Superfund mining communities and beyond. To this end, we will: 1) characterize how inhalation exposure to metal(loid)- containing mine waste particulates lead to lung tissue injury, including compromised mucociliary clearance and epithelial barrier function, tissue disrepair, and fibrosis, focusing on molecular mechanisms, toxicokinetic insights, and identification of individual metal(loid)s in these particulates that are responsible for causing lung injury; 2) characterize interactions between inhaled fungal spores and mine waste particulates in their effects on induction of lung inflammation and injury; 3) develop advanced techniques for the detection, isolation, assessment, and evaluation of mine waste particulates to better understand how mine waste mineral properties influence bioaccessibility, bioavailability, and toxicity; 4) develop advanced technology for assessing, prioritizing, and implementing, phytoremediation in metal-contaminated dryland ecosystems, 5) provide guidelines and tools for targeted phytoremediation solutions at Superfund sites; 6) mitigate the human impacts of exposure to mining waste through effective research translation and community engagement driven by collaborator-engaged research; 7) serve as a global resource for environmental health issues associated with metal mining; and 7) produce transdisciplinary graduates who are equipped with cutting-edge environmental and biomedical expertise to solve pressing hazardous waste problems.

NIH Research Projects · FY 2026 · 1996-09

PROJECT ABSTRACT Focal Adhesion Kinase (FAK) is a multifunctional non-receptor tyrosine kinase and scaffolding protein that is overexpressed in numerous solid tumors including melanoma, while minimally expressed in normal tissue. FAK has been widely investigated as a cancer drug target due its contribution in multiple aspects of tumor progression, including adhesion, invasion, proliferation, survival, metastasis, angiogenesis, and immune cell suppression. However, development of FAK inhibitors has largely focused on inhibition of the kinase enzyme of FAK opposed to inhibition of key scaffolding interactions. Particularly, limited efforts have been made at the discovery and biological evaluation of inhibitors of the focal adhesion targeting (FAT) scaffolding domain of FAK, the domain required for FAK localization to focal adhesions. During this period of support, we have identified the first discovered stapled peptide-based FAK inhibitor (UA-1907) that binds to and co-crystallizes with the FAT domain, and competitively inhibits FAK-paxillin binding. We have identified a myristoylated derivative (UA-2012) with improved cellular potency, favorable drug-like properties, and in vivo efficacy; and developed a bivalent peptide (UA-2023) with low nanomolar binding to FAT. However, the mechanistic differences between these novel FAT domain peptides and traditional FAK-kinase inhibitors on perturbation of focal adhesion complexes and the FAK interactome have yet to be elucidated. Furthermore, we have preliminary data showing that FAT inhibition provides selective anti-cancer effects in NRAS mutant melanoma cells. Melanoma is the deadliest form of skin cancer and there are no current effective targeted therapies against NRAS mutant melanoma, which represents ~30% of all patients. We hypothesize that FAK FAT domain inhibitors have distinct biological effects on the focal adhesion complex in cancer cells compared to FAK kinase inhibitors; and cancer cells with alterations in NRAS signaling pathways have a molecular dependence on FAK FAT scaffolding for survival, thus promoting selective anti-cancer efficacy. In specific aim 1, we will identify the unique differences between FAK FAT scaffold inhibitors and FAK kinase inhibitors on modulation of the focal adhesion complex and the FAK interactome. In specific aim 2, we will define the molecular mechanisms of sensitivity/resistance to FAK FAT inhibition in melanoma cells with activating mutations in NRAS and BRAF pathways. In specific aim 3, we will evaluate in vivo efficacy of novel FAK FAT domain inhibitors in mouse models of NRAS and BRAF driven cancer. Overall, in this project we will utilize FAT stapled peptides to validate that FAT domain targeting provides additional biological efficacy on the focal adhesion complex compared to FAK- kinase inhibitors and that NRAS mutant melanoma has a unique sensitivity to FAK FAT inhibition. .

NIH Research Projects · FY 2024 · 1993-01

To function normally, all cells must maintain ion homeostasis and regulate water content. The lens is unusual because it is made from a packed mass of fiber cells that are incapable of independently maintaining ion and water homeostasis. The fiber cells rely on ion transport mechanisms in a monolayer of epithelial cells at the lens surface. Na,K- ATPase and NKCC1 activity are particularly important. To monitor and control this arrangement, the lens has come to rely on exquisitely specialized remote control mechanisms that utilize TRPV4 and TRPV1 channels. A TRPV4 feedback loop senses swelling in the fiber mass and increases Na,K-ATPase activity to compensate. A TRPV1 feedback loop senses shrinkage in the fiber mass and increases NKCC1 activity to compensate. The feedback loops are important. They explain homeostatic regulation of lens ion transport as well as intracellular hydrostatic pressure, and they fit with the Mathias model of lens circulation. TRPV4 and TRPV1 appear to be master controllers of lens homeostasis. The specific aims are: (1) Test the hypothesis that the TRPV4/hemichannel/Na,K-ATPase response to swelling stretch involves a functional link between TRPV4 and the actin cytoskeleton; (2) Test the hypothesis that the TRPV1/ERK/NKCC1 response to shrinkage involves a functional link between TRPV1 and the tubulin cytoskeleton; (3) Explore reserve mechanisms of lens ion and water homeostasis. Aims 1 and 2 focus on unanswered mechanistic questions regarding TRPV4 and TRPV1 activation by opposing mechanical stimuli, TRPV4-dependent hemichannel opening, and the mechanism of NKCC1 activation. Aim 3 follows up pilot studies on reserve mechanisms that support slower homeostatic responses or serve as a fail-safe backup. The proposed studies are highly significant as regards human vision because preservation of lens transparency and refractive index gradient depends on ion and water homeostasis.