University Of Arizona

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Cigarette smoking is the leading cause of death accounting for more than 480,000 deaths each year, and 16 million Americans suffer from one or more diseases caused by smoking. Cessation can significantly reduce the risk of disease even for those who have smoked for decades. Interventions using a combination of behavioral approaches that focus on maintaining smokers' motivation to quit, providing specific techniques for quitting and relapse prevention, and using Nicotine Replacement Therapy (NRT) are effective at helping tobacco users quit. However, standard behavioral treatment may not appeal to, or be effective for, some smokers and novel approaches are needed to assist these individuals. Guided Imagery (GI) is a form of mind-body therapy that involves controlled, multi-sensory visualization of specific mental images. GI is an effective therapeutic tool to change behaviors including tobacco use. Multiple studies have shown that exposure to GI results in significantly increased abstinence rates compared to controls or those taking bupropion. A large proportion of the U.S. population uses integrative health approaches. A GI tobacco cessation intervention could appeal to smokers not interested in using a behavioral approach. Although GI is an effective tool for smoking cessation, the mode of delivery has generally been in person, limiting dissemination to large populations. Systematic and meta-analytic reviews have shown that telephone quitlines are a highly scalable way to help individuals quit smoking. Quitlines are available in all 50 U.S. states, Puerto Rico and Guam. The proposed project is highly responsive to PAR-20- 154 by delivering a GI intervention via telephone which is highly scalable and could greatly increase the reach and accessibility of an effective GI smoking cessation intervention. We recently completed a randomized feasibility trial of the Be Smoke Free program. Although not powered to determine efficacy, the intervention showed promise in helping participants to quit and showed high consumer satisfaction. We developed procedures for recruiting participants and surpassed our recruitment goals. We had >90% retention at 8-weeks and >80% at 6-months. We also found high levels of adherence. The results of this randomized feasibility study indicate that a fully powered study of a GI tobacco cessation intervention delivered via telephone is warranted. Therefore, the objective of this R01 application is to conduct a randomized controlled trial to test the efficacy of the Be Smoke Free, telephone-based, GI intervention (IC) for smoking cessation compared to active behavioral control (CC). The study will recruit 1,200 diverse smokers from three states, Arizona, New York, and West Virginia to increase generalizability. Participants will be randomly assigned to receive either the IC or CC delivered by telephone by University of Arizona study coaches and will be assessed at 3- and 6-months post-enrollment by study staff. The primary outcome is biochemically verified 7-day point prevalence abstinence at 6 months. This innovative and rigorously designed project conducted by an experienced team has the potential to improve public health through the delivery of an innovative integrative GI intervention via telephone.

NIH Research Projects · FY 2025 · 2021-09

Project Summary The current SARS-COV2 pandemic has brought to light that more efforts are needed to evaluate the pandemic potential of viruses that can spill over in human populations. To assess the pandemic potential of specific viruses, over the next five years my lab will ask if similar viruses caused epidemics not only during the recent documented past, but during the much longer time scale of human evolution. Viruses that caused epidemics in the past are indeed the most likely to cause epidemics again in the future, and hundreds of viral epidemics likely plagued human populations during their evolution. This work will fill gaps in knowledge on epidemics in ancestral human populations, and by doing so, will enable a better assessment of the viruses that represent a future pandemic threat. To study ancient epidemics, my lab will exploit host genomic adaptation driven by ancient viruses. Arms races with viruses have shaped the host immune system by driving a large number of adaptations. I recently showed that viruses left abundant signals of adaptation not only in immune genes, but across the entire human genome. The lab will examine signals of adaptation left by specific viruses in human genomes, to detect, date, and functionally characterize ancient epidemics. To this aim, we will develop new statistical tools based on recent advances in machine learning and in the reconstruction of Ancestral Recombination Graphs (ARGs). These new approaches with increased power to detect and date genomic adaptation will allow us to ask the following questions: 1) Which viruses drove ancient epidemics in human evolution? My lab will create deep learning tests with high power to detect complex genomic adaptation within the past ~200,000 years of human evolution. 2) When did specific viruses drive ancient epidemics? We will use ARGs and Approximate Bayesian Computation to date ancient epidemics, by dating the host adaptive events driven by specific viruses. 3) Which functional host genetic changes were selected during ancient epidemics, in which genes, and how do they influence genetic susceptibility to present viruses? We will investigate regulatory adaptation to viruses and the overall impact of virus-driven host adaptation on the genetic susceptibility of different human populations to specific present viruses, thereby providing virologists with strong candidate host genes for further inquiry. My lab is uniquely suited to decipher ancient epidemics by linking host-pathogen interactions together with the latest developments in the population genomics of adaptation.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Electrical stimulation of deep brain structures is an essential tool for the causal investigation of neural systems that regulate learning and decision making. Deep brain electrical stimulation is also a valuable tool for treating neurological disorders such as Parkinson's disease and tremor, and recent data suggests that electrical brain stimulation may effectively treat epilepsy and severe depression. Despite its scientific and translational applications, little is known about how electrical stimulation affects the ongoing activities of neurons or the release of neuromodulators such as dopamine. Understanding how electrical stimulation affects dopamine release is particularly important given dopamine's involvement in learning, motor control, decision making, and neuroplasticity. There is considerable evidence that dopamine's function is determined by the time course of release. For example, fast, “phasic” release (~1-2 seconds) is involved in neuroplasticity and reward-guided learning while slow, “tonic” release (tens of seconds) is involved in motor control and motivation. Little is known about how the brain selectively regulates tonic and phasic release, and few methods exist for controlling the time course of dopamine release. Developing such methods could result in 1) new experimental approaches for the causal investigation of the roles phasic and tonic release play in motivation and motor control, and 2) translational tools to correct disrupted patterns of dopamine release in disorders such as Parkinson's disease, schizophrenia, addiction, and depression. Towards these goals, evidence from our group suggests that the frequency and duration of electrical brain stimulation allows selective control of the time course of dopamine release. Our general objective is to characterize how parameters of brain stimulation such as stimulation frequency, variability, and the brain region targeted impacts the time-course of dopamine release and dopamine's role in reward-guided learning. Our experimental objectives are to determine (1) how the frequency and variability of the sequence of pulses delivered during brain stimulation affects phasic and tonic dopamine release, (2) how brain stimulation and tonic and phasic signaling interact to affect reward-driven learning, and (3) and how tonic and phasic signaling affect interactions between neurons and shape neuroplasticity. Our experimental approaches involve voltammetry for dopamine measurement, neural ensemble recordings for measurement of neural coordination, optogenetics, and on-line optimization of dopamine release in anesthetized and behaving rats. Our computational objective is to use data collected to develop multi-scale systems and cellular models that describe how stimulation frequency and variance affect the time course of dopamine release. We predict that multi-scale models will outperform current synaptic models and improve the capacity of scientists and clinicians to control dopamine release in experimental and therapeutic settings. These models may also explain clinical effects such as recent data from human patients suggesting that electrical stimulation of deep brain regions reduces depression.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Posttraumatic stress disorder (PTSD) among military Veterans is a critical public health concern. Veteran suicide rates exceed those of the general population, with the disorder creating a mental health challenge that is costly and debilitating. The majority of Veterans with PTSD also have comorbid mental health diagnoses, such as generalized anxiety disorder, substance abuse disorder, and major depression. The treatment of Veteran PTSD and comorbid disorders represents an important therapeutic and rehabilitation problem. The disorder is complex and difficult to treat, with high treatment dropout and nonresponse rates spurring some Veterans to seek complementary integrative health strategies. One promising complementary strategy is the provision of a trained service dog. Initial evidence across multiple research groups highlights service dogs as a promising complement to evidence-based practices that can offer short-term improvements. However, the long-term effectiveness, mechanisms of action, and moderators of efficacy remain largely unknown. Thus, the overarching objective of this proposal is to understand how, why, and for whom PTSD service dogs are most effective. To address this objective, the present project will assess the longitudinal efficacy and dose-response curve of service dogs for Veteran PTSD symptomology and psychosocial functioning (Aim 1). To understand how and why the intervention works, this project will also implement theory-driven quantification of potential mechanisms of action that may mediate service dog efficacy (Aim 2). Finally, to understand for whom and under what circumstances the intervention works best, this project will define moderators of service dog efficacy by examining the heterogeneity of treatment effects (Aim 3). The research design will consist of a two-arm, randomized clinical trial (RCT) with longitudinal assessments at 0, 6, and 12 months. Assessments will consist of comprehensive monitoring across six data streams, including: (1) blinded clinician assessment of PTSD symptomology, (2) standardized surveys of psychosocial functioning, (3) ecological momentary assessment of daily emotional experiences and activities, (4) salivary biomarkers of two major stress response systems via cortisol and alpha-amylase, (5) physical activity and sleep via actigraphy-based wristband monitoring, and (6) canine assessments of behavior, temperament, and physiological co-regulation with the human partner. Results are expected to elucidate the clinical impact of service dogs for military Veterans with PTSD, as well as the biobehavioral mechanisms of action and characteristics that moderate efficacy. These outcomes will support the long-term goal of accelerating complementary and integrative health interventions, through optimized and evidence-based service dog interventions. As such, this project will further advance the scientific understanding of human-animal interactions for psychosocial health.

NIH Research Projects · FY 2025 · 2021-09

Abstract The U.S. faces profound racial and ethnic disparities in hypertension, which are exacerbated by disparities in medication adherence. Yet few existing medication adherence interventions are tailored to address the specific cultural beliefs of diverse populations who also face significant structural barriers to adherence associated with poverty, access to care and food insecurity. The proposed Improving Medication Adherence with Pharmacist and CHW Team (IMPaCT) intervention responds to the call for adherence interventions that are tailored for the specific needs of minority racial-ethnic groups. IMPaCT is a practice-based randomized controlled trial (RCT) to test the effectiveness of a comprehensive, individually- and culturally-tailored intervention for high-risk patients with hypertension, polypharmacy and low adherence. IMPaCT leverages the specialized expertise of a clinical pharmacist together with a community health worker (CHW) who will serve as cultural broker and patient navigator to address individual, clinical, social-cultural, and structural barriers to adherence. This comprehensive and tailored, coordinated care intervention aims to improve medication adherence and hypertension outcomes among African-American, Latino and Vietnamese immigrant patients. The proposed practice-based RCT is designed to meet the following specific aims: Aim 1: Implement IMPaCT, an innovative, tailored adherence intervention delivered by a pharmacist and CHW team. Aim 2: Determine the short- and long-term effectiveness of IMPaCT by assessing pre- to post-intervention changes in: a) medication adherence (proximal outcome) and blood pressure (BP, distal outcome), and b) other comorbid health outcomes (e.g., HbA1c, BMI) using a randomized controlled trial. Aim 3: Identify factors associated with IMPaCT effectiveness including: a) tailored intervention features, b) medication beliefs, c) barriers to adherence, d) intervention dose, e) health literacy, and f) cultural group. We will follow an intention-to-treat randomized design using a waitlist control with 450 African-American, Latino, and Vietnamese patients with hypertension and low (<80%) medication adherence. Data collection via pill count, self-report, the electronic health record, and clinical measures will assess medication adherence, BP and other factors at baseline (pre-intervention) and at 4, 10, and 24 weeks post-intervention. IMPaCT is an innovative coordinated care team intervention to improve medication adherence and blood pressure derived from research findings that builds on existing clinical practice. Designed with an eye towards sustainability, IMPaCT incorporates billable pharmacist and CHW services for patients with low medication adherence and high burdens of chronic illness and preventable consequences. IMPaCT offers interprofessional team care with comprehensive expertise and complementary skill sets that mitigate the silo effect of specialized medicine to deliver primary care to diverse, high-risk populations experiencing disparities in hypertension.

NIH Research Projects · FY 2025 · 2021-09

Nerve injury can affect anyone and carries consequences that last forever, and traumatic peripheral nerve injury (TPNI) can result from settings as diverse as battlefield or motor vehicle trauma, to iatrogenic injury from surgical treatments of other serious conditions. For other tissue types, the prognosis and treatment plan are straightforward. However, when nerves fail, doctors have few tools to help guide the treatment plan and patients forever remember their experiences with pain and paralysis. If nerve injuries can happen from many causes, what is the determinant that most predicts a good outcome? The answer is nerve continuity. A nerve that has been severed, say by a bullet will never heal without surgical repair and yet, that same nerve, grazed by the bullet, or caught in the shockwave of a passing bullet, is indistinguishable from the nerve cut by the bullet. Both nerves do not work, and yet the latter, unsevered nerve is best left to recover on its own without surgery. Two types of nerve injury exist, both yield nerves that are equally dysfunctional and yet we need to surgically repair one whereas surgery is detrimental to the other. No test exists to tell these nerves apart until weeks have passed after the injury. After weeks have passed, we use electrodiagnostics (EDX) to tell which nerves were severed. This is the state of the art in TPNI diagnosis. Because continuity of the nerve is itself impossible to define for weeks after injury, the current standard treatment is waiting for spontaneous recovery in most patients we guess do not have a severed nerve. In every field of medicine, there are nerve injuries monitored in this way - without tools for definitive diagnosis, based on our clinical suspicion. We recently discovered that a current, FDA-approved drug (4-aminopyridine, 4AP), has the previously unknown effect of immediately allowing diagnosis of traumatically paralyzed nerves. Small doses can distinguish the severed nerve, which requires immediate surgery, from the non-severed nerve, which can be safely monitored. This novel property of 4AP, a drug already used in humans may be explored while we wait for function to spontaneously return. Based on our preliminary data, we intend to repurpose 4AP to diagnose nerve discontinuity instead of just guessing if the nerve is severed and hoping for recovery. We want to try 4AP in patients who have nerve injury where we cannot tell if the nerve was severed or not. We obtained FDA approval for a trial of 4AP in humans to investigate nerve continuity and have modified our institutional IRB approval to proceed. We want to test 4AP in humans with TPNI in a novel way: As a pharmacologic diagnostic, where the results of a challenge with the drug reveal something about the underlying nerve continuity. With the knowledge gained from this trial, we hope to provide solid information that can confirm or refute clinical guesswork on the critical issue of nerve continuity.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY/ABSTRACT There is growing need to understand mechanisms and treatment options associated with vascular contributions to Alzheimer’s disease (AD) and other forms of dementia and cognitive dysfunction. Early brain changes associated with AD and other forms of cognitive dysfunction, such as white matter lesion (WML) burden, hypoperfusion and metabolic mismatch have vascular risk factors (hypertension, diabetes, aging and smoking). Such structural and functional brain changes are often considered secondary to intracranial cerebral small vessel disease. In contrast, our preliminary data indicate that extra-cranial carotid artery disease (ECAD) contributes to brain pathology and its treatment with carotid endarterectomy (CEA) (to surgically remove the plaque) decreases accumulation of WMLs and neurofibrillary tangles, increases structural connectivity, and most importantly, improves cognition. ECAD primarily signifies atherosclerosis of the carotid bifurcation, where plaque accumulation is uniquely prevalent compared to other cerebrovascular locations. Carotid arteries play a major role in brain physiology because they bring the majority of blood and nutrients to the brain. We hypothesize that mechanisms of ECAD contribution to Alzheimer’s disease and cognitive dysfunction are likely multifactorial and include embolic phenomena, decreased blood flow, and endothelial activation/inflammation. Interplay between these modifiable factors and non-modifiable risks (ie, age, sex and ApoE status) likely contribute to neurodegeneration that can lead to cognitive dysfunction. Our Aims 1 and 2 use cross-sectional studies to evaluate potential mechanisms of ECAD contribution to AD-related brain structure and function changes. Subject evaluation includes neurocognitive testing and quantification of MRI-defined structural parameters, WML accumulation, Alzheimer’s disease blood-based biomarkers, and systemic, cerebral and carotid markers of inflammation. Aim 3 employs a prospective, controlled cohort study evaluating the treatment of ECAD with CEA to determine which patients show improved cognitive function (responders) and what factors are drivers of this response. This project will define quantifiable measures of brain structural changes leading to neurodegeneration and cognitive dysfunction in subjects with ECAD. This should further build the impetus for clinical trials that change management of patients with ECAD. In addition, our study offers the exciting opportunity to reveal novel insights of early Alzheimer’s disease risk and specific mechanisms of vascular contributions. Understanding the unique contribution of ECAD to AD/cognitive dysfunction risk is particularly compelling because effective treatments exist for ECAD yet are not currently offered for the treatment/prevention of cognitive dysfunction.

NIH Research Projects · FY 2025 · 2021-08

We propose to establish a Center of Excellence for Addiction Studies (CEAS) that will offer core services allowing users to develop projects that will lead to new research in addiction. Addiction and relapse are characterized by dysregulation of brain circuitry that involves diminished activity of brain reward circuits, increased responsiveness of stress circuits and impaired functioning of executive cortical circuits. Neural changes are observed in the basal ganglia, extended amygdala and prefrontal cortical regions and encompass a wide range of endogenous neurotransmitters including dopamine, opioid peptides, endocannabinoids, corticotropin releasing factor (CRF), dynorphin, glutamate and others. While chronic pain and addiction are different disorders, there is a remarkable overlap between the influence of drugs of abuse and chronic pain on these circuits. Our faculty has broad expertise in evaluation of mechanisms that underlie the maladaptations promoted by pain in these circuits. The CEAS will be composed of four Cores and a Pilot Research Project. The Administrative Core will provide the structural elements that will allow efficient functioning of the CEAS. The Genetic Targeting of Neural Circuits Core will allow users to employ cutting edge genetic techniques including CRISPR/Cas9 gene editing, chemogenetics and optogenetics to produce cell and circuit-specific manipulations to evaluate potential mechanisms relevant to addiction. The Neuroanalytical Core will provide users with advanced methods of measuring neurotransmitters with temporal resolution spanning milli-seconds to days and with spatial specificity through advanced detection methods. The Behavioral Core will allow users to explore questions relevant to addiction using behavioral assays that evaluate addictive processes including the influence of addictive drugs on cognitive function. Investigators in the CEAS have worked together for many years and have shared and individual research funding. Additionally, the CEAS will offer opportunities for other investigators at University of Arizona as well as Arizona State University, Northern Arizona University, The University of New Mexico and Texas Tech University Health Sciences Center at Lubbock and El Paso establishing a Southwestern region engaged in addiction sciences. The CEAS will promote increased diversity in addiction research by recruiting investigators and students from under-represented populations in neuroscience and addiction. The impact of the CEAS will be to leverage established funding to develop new research on addiction research. In addition, the impact of funds from the CEAS will be amplified by commitments of matching funds from the University of Arizona and from a recently established Comprehensive Center of Pain and Addiction. The CEAS will provide key services to its users that correspond with the goals of the NIDA to enhance addiction research with a goal of development of therapies that can stem the opioid epidemic as well as impacting other substance abuse disorders. The close collaboration between the Cores ensures high expertise in all areas of the addiction research and will permit outcome measures emphasizing scientific rigor and reproducibility.

NIH Research Projects · FY 2025 · 2021-08

Neuroendocrine tumors arise in multiple tissues, including the skin, lung, prostate, and pancreas, and share common neural and endocrine phenotypes. Discovering their mechanism of carcinogenesis is challenging due to our inability to directly observe tumor formation, as well as the mutational heterogeneity of established tumors. Unlike tumors that arise through random mutational processes, Merkel cell carcinoma (MCC) is a neuroendocrine skin cancer that can be caused by infection with Merkel cell polyomavirus (MCPyV) and whose mechanisms can be studied in vitro. MCPyV contains two oncogenes, the small T (ST) and large T (LT) antigens. The impact of ST on cell migration and LT on the cell cycle are known, but there is no genome-wide, quantitative understanding of how ST and LT reprogram normal cells into a neuroendocrine tumor prone to metastasis. Our preliminary RNA-sequencing data from normal cells expressing MCPyV oncogenes indicates that LT transcriptionally alters invasion and migration-related pathways such as WNT and TGF-beta signaling, in a manner that is similar to expression changes in human MCC tumors and metastases. Using this system, we can also measure gene expression dynamics and directly observe the temporal sequence of molecular events underlying carcinogenesis in a way that cannot be achieved through static tumor measurements alone. The viral etiology of MCC therefore provides us with a unique opportunity to systematically investigate and quantify the regulatory circuits driving neuroendocrine tumor formation and metastasis. In Aim 1, we will reconstruct the host transcriptional networks and signaling pathways utilized by MCPyV to promote cell motility and invasion, determine the roles of LT, WNT, and TGF-beta in promoting metastasis, and predict the optimal strategy for inhibiting these mechanisms. In collaboration with metastasis experts at the University of Arizona, we will test our predictions in a series of MCC model systems, including cell-based migration and invasion assays and mouse xenograft studies. In Aim 2, we will measure protein dynamics of neural regulators activated by ST and LT and integrate them with gene expression and ATAC-seq data to build a network model of the cell fate transition induced by MCPyV. Our work will provide a systems-level analysis of regulatory circuits underlying metastasis and cellular reprograming in neuroendocrine tumors, and will identify new therapies for Merkel cell carcinoma that may be applicable to other tumor types.

NIH Research Projects · FY 2025 · 2021-08

Oral and oropharyngeal squamous cell carcinoma (OSCC) together rank as the sixth most common cancer worldwide, accounting for 400,000 new cancer cases each year. Two-thirds of these cancers occur in low- and middle-income countries (LMICs). While the 5-year survival rate in the U.S. is 62%, the survival rate is only 10- 40% and cure rate around 30% in the developing world. The poor survival rate in LMICs is mainly due to late diagnosis and the resultant progression of disease to an advanced stage at diagnosis. Therefore, it is imperative to diagnose precursor and malignant lesions in LRS early and expeditiously. To meet the need for technologies that enable comprehensive oral cancer screening and diagnosis in low resource settings (LRS) to identify the suspicious lesions, triage the high-risk subjects and thereby enable appropriate treatment management and follow up, this project brings together an interdisciplinary team with complementary expertise in optical imaging, oncology, deep learning, technology translation, and commercialization. The team will develop, validate, and clinically translate a multimodal intraoral imaging system for oral cancer detection and diagnosis with better sensitivity and specificity. This work will address key barriers to adopting optical imaging techniques for oral cancer in LRS by building on the team’s experience in 1) developing and evaluating dual-mode (polarized white light imaging [pWLI] and autofluorescence imaging [AFI]) mobile imaging probes; 2) evaluating a low-cost, portable optical coherence tomography (OCT) system for oral cancer detection and diagnosis in a nodal center setting in India; and 3) developing and evaluating deep learning-based image classification algorithms for clinical decision-making guidance. As each of these key techniques has been demonstrated separately for oral cancer imaging in LRS, the potential of successfully developing a multimodal intraoral imaging system for accurate, objective and location-resolved diagnosis of oral cancer and transitioning to a new capability to medical professionals in LRS is very high. To achieve the project objective, the team proposes three Aims: 1) develop a portable, semi-flexible, and compact multimodal intraoral imaging system; 2) evaluate the clinical feasibility of the prototyped intraoral imaging system and develop deep learning-based image processing algorithms for early detection, diagnosis, and mapping of oral dysplastic and malignant lesions; and 3) validate the capability of the prototyped intraoral imaging system for diagnosing oral dysplasia and malignant lesions. Successful completion of this project will lead to the transition of a multimodal intraoral imaging system and deep learning image classification that leverage the individual strengths of multiple technologies and deliver new and urgently-needed capabilities to the end users in LRS. This integrated system will 1) detect suspicious regions with high sensitivity and specificity; 2) triage the high-risk subjects; and 3) guide the selection of biopsy sites and map lesion heterogeneity to improve treatment planning and intra-operative guidance.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Warfarin remains one of the most commonly prescribed drugs and a leading cause of emergency hospitalizations. Warfarin use is especially common in medically underserved patients such as African Americans (AAs) and Latinos, which is particularly concerning since AAs and Latinos suffer worse outcomes due to suboptimal warfarin therapy. Thus AAs and Latinos can derive a distinct benefit from warfarin pharmacogenomic (PGx) algorithms, which maximize safety and efficacy by predicting individualized warfarin dose. However, currently available PGx algorithms have critical limitations, including a lack of generalizability to non-white populations and a failure to account for 50 percent of variability in warfarin dose. Under-representation in clinical studies, the propensity to cause adverse events, and a lack of consideration of admixed populations in clinical PGx guidelines are all factors that contribute to limited utility of warfarin PGx algorithms in diverse populations. Many potential sources of warfarin stable dose variability remain critically unexplored, including the role of vitamin K biosynthesizing bacterial species, the influence of local ancestry at warfarin pharmacogenes, and the potential for machine learning techniques to enable accurate warfarin dosing algorithms in diverse populations. This proposal addresses the overarching hypothesis that warfarin stable dose prediction can be improved by incorporation of gut microbiome data, measures of local ancestry, and machine learning in diverse populations. We will pursue three Specific Aims (SAs) to test this hypothesis: (SA1) Determine the impact of abundance of vitamin K biosynthesizing bacteria from the gut microbiome on warfarin stable dose and; (SA2) Determine the influence of local admixture on warfarin stable dose in admixed populations; (SA3) Optimize warfarin PGx algorithms for diverse populations using machine learning. In SA#1, we will conduct a clinical study with fecal sample collection at anticoagulation clinic visits and perform whole genome bacterial sequencing to identify the effect of vitamin K biosynthesizing bacterial species on warfarin stable dose. In SA#2, we will estimate African, European, and Native American local ancestry in warfarin pharmacogenes in a large, admixed population (n=1194) and determine its effects on warfarin stable dose. In SA#3, a large, diverse population of warfarin treated patients (n=7366) will be used to develop machine learning models and test improved prediction of warfarin stable dose over existing linear regression models. Our studies overcome major limitations of previous warfarin PGx studies by leveraging gut microbiome data, local ancestry, machine learning, and diverse, admixed populations. The outcomes of this work will provide a framework for local ancestry investigation with other PGx drug-gene pairs, enabling use of clinical PGx guidelines in admixed populations. This research has the potential to identify new sources of variability in warfarin dose, improve the safety and efficacy of warfarin treatment, and reduce disparities in PGx research for medically underserved patients.

NIH Research Projects · FY 2025 · 2021-08

Project Summary: Many patients suffer from chronic pain in the absence of identifiable injury. Such pains are termed “functional” and include irritable bowel syndrome, temporomandibular joint disorder, fibromyalgia, migraine and others. For reasons that are not understood, almost all functional pain syndromes (FPS) are female prevalent. FPS patients experience pain-free interictal periods punctuated by attacks of pain. The frequency of attacks is predictive of risk of chronification. Pain episodes thus produce a priming effect, establishing a state of increased vulnerability to future attacks, likely reflecting peripheral and central sensitization. FPS patients commonly identify stress as a key trigger of pain. Repeated stress may thus promote vulnerability and pain in a sexually dimorphic fashion. We have developed an injury-free rodent model of FPS based on hyperalgesic priming with repeated stress. Hyperalgesic priming produces a pain-free state of increased vulnerability that has been termed “latent sensitization” (LS). Following induction of LS, normally subthreshold triggers can produce pain attacks, modeling the interictal and ictal periods of FPS. We will use this model to test the novel hypothesis that repeated stress activates kappa opioid receptor (KOR) signaling in the hypothalamus resulting in release of prolactin (PRL) and dysregulation of prolactin receptor (PRLR) isoform expression selectively in female nociceptors. PRL signals through homodimers of PRLR long and short (i.e., PRLR-L and PRLR-S) isoforms that respectively regulate transcription and pain. Repeated stress down-regulates PRLR-L promoting female-selective pain through stress-induced PRL/PRLR-S signaling. The balance of PRLR isoforms may therefore “tune” female nociceptors to promote LS and pain from normally subthreshold stimuli. We will use genetic and chemogenetic manipulations along with anatomical, neurochemical, electrophysiological, pharmacological and behavioral studies in male and female mice to evaluate the role of dorsal root ganglion (DRG) PRLR-L down-regulation and stress-related hypothalamic KOR activation as essential mechanisms of LS and stress-related pain in females. Aim 1 will establish the effects of repeated stress on hypothalamic KOR signaling and PRL release. Aim 2 will establish a potential causal relationship of repeated stress or hypothalamic KOR activation on DRG PRLR isoform expression, neural excitability, LS and stress-related pain. Aim 3 will determine if KOR antagonists, DA agonists or a PRL antibody will prevent LS and FPS-like pain selectively in females. The proposed studies will characterize a previously unknown stress-related neuroendocrine link between hypothalamic KOR and PRL/PRLR signaling to promote female selective functional pain. Importantly, these studies will advance knowledge about previously unknown biological mechanisms and may unravel mechanisms for therapeutic interventions allowing improved therapy of FPS in women.

NIH Research Projects · FY 2024 · 2021-08

Project Summary / Abstract Autism spectrum disorder (ASD) is a developmental disorder that affects 1 in 54 children in the US (1). The economic cost of ASD is estimated to be $66 billion per year in the US, from medical care and lost parental productivity (2). Early diagnosis is crucial since it allows for early treatment and the best long-term outcome. However, identifying children at high risk for ASD at an early age is challenging due to lack of specialists. To address this problem, the project's objective is to create health information technology (HIT) using information in electronic health records (EHR) to support non-expert clinicians in identifying children at high risk for ASD. The HIT will integrate two components that provide complementary information. The first component will leverage machine learning algorithms to label EHR of children at high risk for autism. Both traditional and deep learning, potentially leveraging each other, will be evaluated while systematically tracking quality and quantity of information in EHR and their effect on performance. The second component will focus on the EHR free text and identify phenotypic behavioral expressions of diagnostic criteria as defined in the Diagnostic and Statistical Manual of Mental Disorders (DSM). Rule-based natural language processing will be combined with machine learning algorithms. For both components, potential algorithm bias will be investigated and corrected or documented when this is not possible. The HIT will combine results from both components through an intuitive user interface. Since it is intended to be used as a human-in-the loop decision tool, it will also provide descriptive data on performance for both components. The final HIT will be developed using rapid prototyping in interaction with domain experts. It will be evaluated in a user study with representative non-expert clinicians. The evaluation will compare accuracy, confidence, and efficiency of identifying children at risk for ASD with and without the HIT by non-ASD experts. It will also systematically focus on the type, amount, quality and transparency of information provided, and how this interacts with user beliefs about their own expertise as well as their bias toward machine decisions. Different types of EHR as well as different levels of clinical expertise will be compared for effects of HIT use. Preliminary work has been conducted for all components with good results. However, this prior work focused on version IV of the DSM and used only free text from data rich EHR. The proposed project will expand the prior work to use DSM-5 criteria, train and develop the algorithms to use structured and unstructured fields in clinical, representative EHR, and work with EHR from different hospitals to evaluate potential obstacles and advantages of variability in data. Using information in EHR, this HIT will provide support especially for non-expert clinicians in their evaluation of children who may be at risk of ASD. The HIT will support early referrals leading to early diagnosis and therapy. It will be useful in a variety of different settings where domain expertise is missing.

NIH Research Projects · FY 2024 · 2021-08

Frontotemporal dementia (FTD) is an age-related non-Alzheimer dementia characterized by progressive neuronal loss in the frontotemporal lobes. A subset of FTD is defined by the pathology of protein inclusions positive for Fused in Sarcoma (FUS), thus named FUS-FTD. FTD and amyotrophic lateral sclerosis (ALS) share a wide spectrum of clinical, pathological and genetic features. Pathogenic mutations of FUS cause both ALS and FTD, and FUS proteinopathy is also detected in sporadic diseases. FUS is primarily in the nucleus, but the protein with a disease-causing mutation mislocalizes and accumulates in the cytoplasm. Protein aggregates induced by FUS mutations may sequester other proteins to compromise cellular functions, resulting in impaired neurons. We recently showed that FTD/ALS mutations of FUS suppressed protein translation and hyper-activated the nonsense-mediated decay (NMD) of mRNAs. The hypothesis to be tested in this project is that the dysregulation of protein translation and mRNA surveillance contributes to cortical neuron loss in FUS-FTD. Three specific aims are designed to test the hypothesis using in vitro and animal models as well as FTD patient tissues. Aim 1 is to determine how mRNA NMD and protein translation are perturbed by pathogeneic FUS mutations in FTD mouse models and patient tissues. We will determine whether NMD factors and translation-related proteins are sequestered in FUS inclusions in forebrain neurons in R521G FUS transgenic mice at different ages. We will measure mRNA turnover rates and protein translation efficiency in R521G FUS mice and correlate the perturbations to neuronal dysfunction and FTD disease progression. Moreover, we will examine whether NMD factors and protein translation proteins are sequestered in FTD patient tissues. Aim 2 is to identify specific proteins suppressed by pathogenic FUS in FTD mice. We will apply the puromycin labeling in FTD mice and use the proteomic approach to identify changes in protein translation impacted by mutant FUS in forebrain neurons. In addition, actively translated mRNAs will be identified and quantified in polysome fractions using RNA-Seq. Results from the –omics approaches will be integrated for pathway analysis to reveal whether pathogenic FUS impairs proteins in specific pathways, providing novel insights into the FTD etiology. Aim 3 is to elucidate the significance of RNA binding and post-translational modifications in the dysfunction of pathogenic FUS. We will use a cohort of RNA binding-deficient mutations in an optogenetic Cry2olig-FUS-mCherry system to examine the significance of RNA binding in FUS inclusion formation and dysfunction in cortical neurons. We recently found FUS is acetylated at residues critical to RNA binding, we will test how acetylation-null and -mimicking mutations affect FUS inclusions, NMD and protein translation. In addition, we will also examine whether pathogenic FUS forms RNA-dependent and -independent inclusions that produce different levels of toxicity to neurons. The proposed experiments will thoroughly examine a novel disease mechanism using innovative approaches. Completion of our proposed work will help elucidate molecular mechanisms underlying FUS FTD.

NIH Research Projects · FY 2025 · 2021-07

ABSTRACT The management of chronic pain is clinically challenging, and relies heavily on opioid drugs like morphine and oxycodone. However, opioids are plagued by numerous side effects that impact quality of life, like tolerance, constipation, and reward/addiction, contributing to an opioid abuse, addiction, and overdose crisis. These clinical and social challenges highlight the vast medical need for new approaches to pain management. To this end, we have pioneered an investigation into the role of Heat shock protein 90 (Hsp90) in regulating opioid signal transduction, anti-nociception, and side effects. We have found that Hsp90 regulates mu opioid receptor (MOR) signal transduction to different effect in brain vs. spinal cord. In brain, Hsp90 promotes MOR signaling and anti- nociception, so that Hsp90 inhibition in brain blocks opioid anti-nociception. In spinal cord, Hsp90 blocks MOR signaling and anti-nociception, so that Hsp90 inhibition in spinal cord enhances opioid anti-nociception. In further studies, we found that Hsp90 inhibition in spinal cord increases morphine anti-nociceptive potency 2-3 fold in acute and chronic pain, reduces tolerance and rescues established tolerance, all without altering the potency of constipation and reward. These results suggest that spinal Hsp90 inhibition could be used as an opioid dose-reduction strategy, to improve or maintain analgesic efficacy while reducing side effects. However, one challenge to this approach is our finding that non-selective Hsp90 inhibitors, when given systemically, gave results similar to the brain, blocking opioid anti-nociception. Seeking a way around this limitation, we found that Hsp90 isoforms differ between brain and spinal cord, with Hsp90α alone acting in brain while Hsp90α, Hsp90β, and Grp94 all act in spinal cord. Hypothesizing that an isoform-selective Hsp90 inhibitor could be used to target spinal cord-specific isoforms, we found that the Hsp90β-selective inhibitor KUNB106 enhanced morphine anti- nociception while rescuing established morphine tolerance when given systemically. These results strongly suggest that Hsp90β-selective inhibitors could be used as a novel, first-in-class opioid dose-reduction therapy. However, KUNB106 is a first generation compound, with poor solubility and pharmacokinetics (PK) and an uncertain therapeutic profile. In this proposal, we will thus optimize KUNB106 to create a new therapeutic to enhance opioid therapy and reduce opioid side effects like reward/addiction. In Aim 1 we will utilize cutting edge medicinal chemistry approaches using Hsp90 isoform co-crystallized structures to create optimized compounds based on the KUNB106 scaffold. In Aim 2, we will test these compounds for Hsp90 isoform selectivity, ADMET parameters, off-target interactions, and in vivo PK in mice, aiming to identify highly selective, soluble, and orally bioavailable compounds. In Aim 3, we will test the best of these compounds for their efficacy in enhancing opioid anti-nociception in acute and chronic pain models in mice, while reducing tolerance, constipation, reward, and respiratory depression. Top candidates will be tested for off-target side effects and toxicity. Through this project, we aim to create optimized candidates for further development as new therapeutics for patient pain management.

NIH Research Projects · FY 2025 · 2021-07

PROJECT SUMMARY/ABSTRACT: The COVID-19 pandemic has led to dramatic reductions in cancer screening and follow-up services. During the early months of the pandemic, national organizations recommended postponing all routine cancer screening, resulting in dramatic reductions in adult primary care and specialty care visits; current rates remain far below pre-pandemic levels. Modeling suggests that these reductions will result in over 57,000 missed cancer diagnoses and 10,000 excess deaths over the next decade. However, these models are largely informed by data sources made up predominantly of insured patients and lacking race/ethnicity information. Thus, more information is needed to understand the strategies and resources need to support the recovery of health systems and communities that have been most impacted by COVID-19. The effects of the pandemic on cancer screening uptake may be particularly profound for patients served by resource-limited federally qualified health centers (FQHCs), which deliver services a large share of patients with incomes below the federal poverty level and who are Latinx. Latinx populations already have some of the lowest rates of cancer screening and follow-up care in the United States and are likely to experience the largest reductions in care and slower return to normal following COVID-19. This is, in part, because their communities have been hit particularly hard by COVID-19 (e.g. high rates of COVID-19 infections and hospitalizations and job loss) and they may fear returning for preventive care even when medical authorities have deemed it safe. Our proposed mixed-methods study will estimate the impact of the COVID-19 pandemic on rates of cancer screening and follow-up in patients served by a large, diverse FQHC (Aim 1), and estimate impacts on cancer outcomes (e.g. changes in life years gained, cancers prevented, and late-stage cancer incidence) in the FQHC population building on models developed by the CRC-SPN Cancer Intervention and Surveillance Modeling Network (Aim 2). Finally, we will gather qualitative data from clinic staff and patients to identify opportunities to improve post-pandemic cancer preventive care delivery for adults served by FQHCs (Aim 3). There is a critical need to understand the long-term impacts of COVID-19-care reductions on vulnerable populations and identify opportunities to meet the ongoing cancer prevention needs of patients served by FQHCs. We will collaborate with national stakeholders to develop FQHC-specific guidance to inform future interventions to support recovery from COVID-like care disruptions Thus, our findings will support access to care and reduction of health disparities for communities most impacted by COVID-19.

NIH Research Projects · FY 2024 · 2021-07

Summary Chronic epithelial or vascular injuries followed by dysregulated repair are the trigger mechanisms in pathogenesis of interstitial lung diseases, including idiopathic pulmonary fibrosis (IPF). Multiple cell types are involved in lung fibrogenesis, with fibroblasts and epithelial cells given the most attention. Role of endothelial cells and microvasculature remain unclear. Dysregulated repair causes vascular remodeling, associated with increased vessel permeability, partial loss of capillaries, focal increase in pathological angiogenesis and endothelial dysfunction. Normal endothelial cells (EC) are transcriptionally re-programmed into fibrosis- associated endothelial cells (FEC), that support activated fibroblasts and promote lung inflammation. Our long- term goal is to identify key regulators of EC-to-FEC re-programming and clarify molecular mechanisms of the crosstalk between endothelial cells and other cell types during pulmonary fibrogenesis. In our preliminary data, we used endothelial cells from lungs of patients with IPF and mouse lung fibrosis models to identify FOXF1 as a key transcriptional regulator of EC-to-FEC re-programming during lung fibrogenesis. Using transgenic mouse models with endothelial-specific deletion or over-expression of Foxf1 gene, we propose to test the hypothesis that endothelial FOXF1 decreases activation of fibroblasts and prevents macrophage accumulation in fibrotic foci. We propose two specific aims: (1) identify molecular mechanisms whereby endothelial FOXF1 inhibits lung fibrogenesis, (2) establish whether restoring FOXF1 in FECs will prevent or reduce fibrotic lung remodeling after chronic lung injury. Understanding the regulation of EC-to-TEC re-programming, and the molecular mechanisms utilized by pulmonary endothelial cells to control pulmonary fibrosis, will provide new approaches for treatment of interstitial lung diseases.

NIH Research Projects · FY 2025 · 2021-07

Project Summary/Abstract Biocompatible chemical transformations that are promoted by light have become powerful tools in chemical biology by virtue of enabling spatiotemporal control over activity. Whilst genetically encoded photoactivatable tools have become mainstays in the bio-orthogonal toolbox, light driven conjugation methods that effectively interface with native biomolecular structures (no genetic encoding) under biologically relevant conditions, are comparatively limited. In this project, we will evolve a method recently developed by our group for the photobioconjugation of Tryptophan (Trp) residues using redox-active N-carbamoylpyridinium salts that engage Trp in photo-induced electron transfer. We will show that, by carefully modulating the optical and electrochemical properties of these reagents, that we will be able to both (1) dramatically enhance the kinetic capabilities of this labelling reaction and (2) enable the discovery of new mechanistic paradigms that promote this labelling chemistry. Moreover, we will demonstrate that, through careful manipulation of optical and electrochemical properties of the N-carbamoyl pyridinum salt reagent, that we will be able to invoke mechanistic control over Trp labelling in a wavelength-dependent fashion (i.e. we can control reaction mechanism with a given wavelength of light). This, in turn, will allow us to design new application-based experiments that can both command precise reaction outcomes and markedly expand the capabilities of photobioconjugation chemistry. Specifically, we will harness this optical and mechanistic control for the design of new activity-based sensing applications as well as through the design of proximity labelling approaches that we apply to the study of poorly understood processes in mitochondrial dynamics.

NIH Research Projects · FY 2024 · 2021-07

ABSTRACT Parkinson's disease (PD) is the 2nd most common neurodegenerative disorder, affecting over 1 million people in the United States. PD causes difficulties with movements such as walking and speaking that occur because of loss of the brain chemical dopamine. Current symptomatic PD treatments are based largely on dopamine replacement therapies with L-DOPA; however, these treatments have many long-term side effects which led to interest in non-dopaminergic therapies. The most severe side effect is the development of L-DODA-induced dyskinesia (LID), involuntary movements that can be as or even more debilitating than the disease itself. Any adjunct therapy extending the time frame where L-DOPA can be used without LID would be a major advance. Recent publications showed that low-dose ketamine infusion paradigms were safe and well tolerated in clinical trials for pain states (including migraine headaches), treatment-resistant depression and posttraumatic stress disorder (PTSD). Low-dose ketamine has led to a long-term reduction of pain states, treatment-resistant depression, it also reduced PTSD symptom severity and comorbid depression. One commonality between migraine headaches, depression, PTSD, PD and LID is that electric activity in the brain is overly synchronized and maladaptive plastic changes occur in the brain, including in an area that is of interest in PD and LID called the basal ganglia (BG). Therefore, we investigated the use of low-dose sub-anesthetic ketamine in the treatment of PD and LID. We have evidence of a therapeutic effect of low-dose ketamine infusion from preclinical data in a rat model of LID (dose-dependent reduction of abnormal involuntary movements; long-term effects after a single ‘infusion-treatment’) and from 5 PD patient case studies (reduced dyskinesia and reduced depression). In the rat LID model this effect was only seen when low-dose ketamine was given for 10 hours and not with just a single acute low-dose ketamine injection. The premise of the proposed studies to define mechanisms of the novel use of low-dose ketamine is ‘true bench to bedside’ science, will provide the foundation for controlled clinical trials of low-dose ketamine treatment for LID, and could identify new more specific therapeutic drug targets to treat LID and depression, two critical problems for many PD patients. Our main hypothesis is that a low-dose sub-anesthetic ketamine infusion desynchronizes overly synchronous oscillatory activity in nerve cells involved in LID sufficiently to induce a lasting anti-dyskinetic effect, working as a “chemical deep brain stimulation (DBS)”. We hypothesize that ketamine works on the molecular level via 2 types of receptor molecules in the BG and cortex, NMDA receptors and opioid receptors, and that the long-term effect includes changes in nerve cell connections called dendritic spines. A multidisciplinary team of researchers and a clinician with the necessary expertise will fill a critical gap in knowledge by investigating the mechanisms of this long-term effect of low-dose ketamine infusion on the molecular and cellular level. They will study effects on receptors and changes in spine size and density (Aim 1), and on the systems level, investigate synchrony of oscillatory neural activity (Aim 2).

NIH Research Projects · FY 2025 · 2021-07

ABSTRACT Chronic pain is a serious and worsening epidemic in the United States and worldwide, seriously degrading patient quality of life. Opioid drugs like morphine are the “gold standard” for treating moderate to severe chronic pain, however, they are burdened by major side effects, especially addiction liability, which has contributed to a paralell epidemic of opioid addiction, abuse, and overdose. In addition, opioids are ineffective in some pain types, most notably neuropathic pain. In the search for alternatives, phytocannabinoids from Cannabis sativa have been heavily studied. However, cannabinoids have generally been shown to have modest to poor efficacy, and have their own side effects, especially psychoactive side effects with Δ9-tetrahydrocannabinol treatment. This has led again to a search for methods to improve cannabinoid therapy. For this reason, research has focused on the ~150 terpene compounds found in Cannabis, which impart flavor and aroma to the plant. Limited evidence suggests that terpenes produce pain relief on their own, and they have also been proposed to modulate and potentially improve the effects of cannabinoids like THC, termed the “entourage effect” hypothesis. However the quality of evidence on terpene efficacy is in general poor, limited by poorly-defined and complex extracts, and few mechanistic studies. We thus performed a preliminary study on the Cannabis terpenes α- humulene, β-pinene, geraniol, and linalool. We found that all 4 terpenes produced anti-nociception in a mouse model of chemotherapy-induced peripheral neuropathy (CIPN) comparable or better than morphine. At the same time, geraniol and linalool produced no reward or aversion, suggesting no addictive or aversive liability. Seeking mechanistic insight, we found that all 4 terpenes produced tail flick anti- nociception by a cannabinoid receptor type 1 (CB1) mechanism, and further synergized with the cannabinoid WIN55,212, providing evidence for the entourage effect hypothesis. We further identified CB2, Adenosine A2a, and anti-inflammatory activity as potential mechanisms of action. In this proposal, we will extend these studies to evaluate therapeutic potential and mechanisms of action of these terpenes in neuropathic pain, providing potential support to the use of these ligands as improved non-opioid pain therapeutics. In Aim 1, we will fully test the terpenes in a mouse model of CIPN, including dose/response, alternate neuropathy models, side effects like tolerance and reward/aversion, synergy with other analgesics such as opioids and cannabinoids, and terpene impact on side effects of these other analgesics (especially opioid reward). In Aim 2, we will identify molecular mechanisms for terpene action in CIPN, focusing on 1) CB1/2, 2) A2a, and 3) anti-inflammatory activity. We will use selective antagonists and CRISPR gene editing, identify sites of action (e.g. brain, spinal cord, periphery), measure tissue response to terpene (e.g. cytokine production), and use in vitro models to confirm these mechanisms. Together these studies will provide a rigorous evaluation of the potential use of terpenes as efficacious and low side-effect therapeutics for neuropathic pain.

NIH Research Projects · FY 2025 · 2021-07

PROJECT SUMMARY/ABSTRACT As the leading cause of cancer death, lung cancer represents the most significant cancer-related public health challenge in the United States. Although low-dose CT-based screening holds promise for earlier detection, currently, most lung cancer cases are not diagnosed until advanced stages (III, IV) and have 5-year survival rates of 21% or less. In contrast to Black-White survival disparities, Hispanic patients have markedly lower age-adjusted death rates than their non-Hispanic White (NHW) counterparts despite later stage diagnoses and broader SES and healthcare disparities; these findings are well established and consistent with the “Hispanic Health Paradox”, a phenomenon characterized by Hispanic advantages in objective health outcomes (e.g., mortality) despite significant health and socioeconomic risk factors. The leading explanatory hypothesis concerns the role of cultural factors facilitating social integration. Social integration is among the most robust psychosocial predictors of a range of objective health outcomes, including cancer survival. However, no published work has directly tested this sociocultural hypothesis in relation to Hispanic resilience. Consistent with the emerging science of resilience, we propose a multisite, two-study, mixed-methods investigation to evaluate this sociocultural hypothesis. Study 1 is a multisite, longitudinal observational study of 672 Hispanic and NHW individuals with advanced stage lung cancer sampled from three regions across the U.S. Interviews using gold-standard and culturally-informed survey measures (demographics, social integration, cultural values, acculturation) will be conducted in English and Spanish from a centralized coordinating center with a 6- week follow-up to examine change in perceived support provision/needs. The primary outcome of survival and secondary outcomes (e.g., treatment adherence), will be gathered from electronic medical records over mean follow-up time of 33-months. Study 2 is a single-site, 7-day, intensive measurement investigation into the daily units of social integration that mediate outcomes. Study 2 integrates two novel in vivo sampling methods (Electronically Activated Recorder [EAR] and ecological momentary assessments [EMA]) using a mobile phone platform. The current aims are to (1) investigate whether the observed Hispanic survival advantage is mediated by ethnic differences in social integration among recently diagnosed late-stage lung cancer patients and (2) to examine the processes/mechanisms that underlie these relationships in daily life including the role of individual, family, network, and neighborhood-level factors. The highly experienced investigator team includes leaders in all relevant content areas, including the Hispanic health paradox, lung cancer survivorship, social integration, and ecological sampling methodologies. The results will contribute to better understanding of social processes among cancer patients, inform psychosocial interventions based on social integration, and contribute to the emerging science of health resilience as well as racial/ethnic and cultural variations in health outcomes.

NIH Research Projects · FY 2025 · 2021-07

Members of the Multi-drug Resistance Protein (MRP) family of ATP Binding Cassette (ABC) transporters contribute to drug tolerance in major fungal pathogens, including Candida species, in 2 main ways: 1) they detoxify the cell of cytotoxic molecules such as antifungal drugs and electrophiles/oxidants by sequestering them in the vacuole, and 2) they cause complex morphological changes such as hyphal extension and biofilm formation linked to drug tolerance. Because of their role in these survival mechanisms, MRP family members are often required for infection and are tightly regulated by the cell. The overall objective of this proposal is to bridge gaps in our understanding of how these transporters contribute to the increasing threat of drug resistance in fungal pathogens. Mechanistic models that explain this resistance are especially lacking, limiting antifungal treatment efficacy. The long-term goal is to understand how anti-fungal treatments fail. To this end, we specifically focus on early stage anti-stress responses through enzymatic, structural, and cellular investigations of two prominent vacuolar MRP transporters: Ycf1 and Mlt1. Our rationale is that these insights will provide a foundation for exploiting unique aspects of transporter architecture in order to generate more effective therapies that overcome drug tolerance in fungal infections. Our preliminary work identifies key features of MRP family transporter regulation during fungal stress responses. These are complex substrate binding sites and the regulation of different states of an intrinsically disordered region called the Regulatory-domain (R- domain) by phosphorylation. Our central hypothesis is that MRP transporters are governed by domain insertions outside of the conserved transporter fold that regulate multi-segmented substrate binding sites. Our specific aims testing this hypothesis in MPR transporters are to define: (1) the regulatory architectures adopted across transport cycles, (2) the functional and structural roles of R-domain phosphorylation on catalytic regulation, and (3) the molecular basis of substrate selection and lipid transport in membrane homeostasis. This application uses an innovative multidisciplinary approach that applies advances in membrane protein biochemistry and electron cryo-microscopy to structurally and biophysically undercharacterized fungal integral membrane proteins. In light of the growing threat of Candida infections, the significance of our proposal is twofold. First, we will establish a mechanistic framework for understanding the molecular basis of therapeutic failure driven by important Candida virulence factors and their yeast homologs. Second, we will establish a foundation useful for developing allosteric therapeutics against drug-tolerance linked to fungal MRP family transporters, with general applicability to all MRP transporters.

NIH Research Projects · FY 2025 · 2021-07

PROJECT SUMMARY Human cytomegalovirus (HCMV) is a herpesvirus that causes disease and death in the immunocompromised and is a leading cause of congenital disabilities. HCMV replication requires lipids. Since HCMV does not encode a metabolic network, virus replication depends on host lipid metabolism. However, little is known about how HCMV reprograms host metabolism to ensure lipids required for virus replication are made. Our overall goal is to understand the virus-host interactions that regulate lipid synthesis essential for HCMV replication. Recently, we showed that HCMV infection results in an increase in lipid synthesis and a rise in lipid abundances. Here we demonstrate that HCMV infection induces the synthesis of at least 20 previously undescribed lipids unique to infected cells. Most of these unique lipids are phospholipids with very long-chain fatty acid tails (PL-VLCFAs). The PL-VLCFAs discussed in this application are understudied in general and unstudied in HCMV biology beyond our work. While shorter FA tails have been well-studied, we know little about lipids with VLCFAs tails that are as long as those we observe in HCMV infection, including how they will behave in a biological membrane. The molecular mechanisms underlying this HCMV-induced expansion in the host lipidome and the functional roles of the newly generated lipids are largely unknown. We discovered that HCMV pUL37x1 and pUL38 proteins promote PL-VLCFA synthesis, laying the foundation for understanding the mechanisms by which HCMV reprograms lipid synthesis. pUL37x1 and pUL38 induce Ca2+ and mTOR signaling, respectively. We have preliminary data suggesting that stress responses related to these signaling pathways contribute to HCMV remodeling of lipids. We hypothesize that pUL37x1 and pUL38 use Ca2+ and mTOR signaling to promote the synthesis of PC-VLCFAs required for HCMV replication. We will test this hypothesis by determining the mechanisms by which pUL37x1 and pUL38 promote synthesis of PL-VLCFAs (Aim 1) and defining the PL-VLCFA synthesis enzymes required for HCMV replication and the role of PL-VLCFAs in infection (Aim 2). These studies will determine the mechanisms by which HCMV interacts with the host to create a unique lipid environment advancing our knowledge of HCMV reprogramming of metabolism. Furthermore, these studies will define the biological functions of PC-VLCFAs in HCMV replication and further our understanding of lipids required for HCMV infection. Determining the mechanisms involved in HCMV-induced reprogramming of lipid metabolism and functions of PC-VLCFAs will advance knowledge in HCMV biology needed to identify new targets for treating infection.

NIH Research Projects · FY 2025 · 2021-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Our mission within the UA COM-T, MD-PhD Program is to develop a pool of highly trained physician-scientists who have the aptitude to utilize clinical experience in developing biomedical hypotheses that integrate the research skills learned in our program. We intend to have all trainees independent, productive and rewarding physician-scientist careers. This program has established 6 objectives that include; 1) dual-degree completion rates at >97% with an appropriate time-to-degree (7.3 yrs on avg.), 2) the integration of research and clinical activities, 3) nurture a broad understanding of biomedical disciplines, 4) foster and encourage the development of good scientific premise, rigorous-research design, strong experimental methods with reproducibility/validation, as well as skills to analyze and interpret results/outcomes amid ethics and integrity, 5) build proficiency in initiating, conducting, interpreting, and presenting rigorous and reproducible biomedical research with increasing self-direction, while strengthen skills to teach and communicate, and 6) advancing the knowledge, professional skills, and experiences required to identify and transition into productive careers in the biomedical research workforce that utilize the dual-degree training. UACOM-T MD-PhD Program offers a wide range of biomedical disciplines for trainees with 13 graduate programs available and over 60 faculty trained and excited to be mentors. Our rationale for an NIH-MSTP is the desperate need for the increase in Physician-Scientists in the state of AZ; a population that is rapidly growing and aging with multiple medical and research needs. AZ continues to increase its ability to attract both academics and biomedical industries that are advancing the care for those who suffer from things like dementia, arthritis, diabetes and cardiovascular issues. These industries and academic institutions are at great need for training and hiring physician scientists. The University of Arizona’s continual growth in the biomedical sciences places UArizona’s projections as one of the top of all Research I, AAU-member institutions in the southwest. Hence, the UA COM-T is a perfect institution to serve and uphold the goals and intentions of the MSTP by increasing the number of physician scientists. The current MD-PhD program has been funded entirely by support from the institution resulting in a small but overall, very successful cohort of physician scientists with past trainees in academic positions at Stanford, Harvard, Yale, Vanderbilt, Baylor, OHSU, Scrips, etc. Our program has established a comprehensive 7-year training plan that reduces unnecessary redundancies, utilizes evidence-based medicine training, integrates clinical and research over all 7 years while creating and encouraging a vibrant climate.

NIH Research Projects · FY 2025 · 2021-05

Our previous community-based participatory research at Northeast Cape (NEC) on St. Lawrence Island (SLI) found elevated levels of polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), and mercury (Hg) in sediments and biota within the watershed at the formerly used defense (FUD) site. We found elevated OCPs and PCBs in serum of the SLI people due to both long-range transport and military-derived sources, with the highest levels of PCBs in people who have traditional and familial connections with NEC, including subsistence. Concentrations of persistent organic pollutants (POPs) in our resident fish model, the stickleback, closely mirror concentrations in the blood serum of SLI residents, indicating their efficacy as sentinel species on SLI. Despite extensive remediation at NEC, short-lived lower trophic level fish in the Suqitughneq (Suqi) River remain contaminated with PCBs, OCPs, and Hg originating from the FUD site, at levels that exceed EPA fish consumption guidelines for cancer risk. Elevated contaminant levels and disrupted health in Suqi River fish indicate potential health threats for residents and that site remediation is incomplete. Our overarching goal is to advance scientific understanding of the exposure pathways and long-term human health consequences associated with contaminant exposure from FUD sites and inform interventions necessary to protect the health and long-term well-being of the people of SLI as they re-establish their traditional community at NEC. In Aim 1, we propose to build on our prior discoveries and continue our collaboration with the communities of SLI to investigate potential exposure pathways and biological impacts of persistent contamination associated with the FUD site at NEC on SLI. We will analyze PCBs, OCPs, and Hg in the water of the Suqi and Tapisaggak rivers, as well as in traditional foods and air samples to assess ingestion and inhalation as potential exposure pathways. We will build on work with stickleback as a sentinel species to determine biological effects of contaminants on endocrine function and organ-specific histopathology. In Aim 2, we will characterize and quantify body burden of contaminants, and linkages to health outcomes, in people associated with NEC. In Aim 3, we will inform decisions and interventions to protect the health of the people of SLI and enable re- establishment of the traditional community at NEC. We will provide information that will lead to improved remediation, provide data and traditional knowledge to inform public health consultations and assessments, and develop a community-based public health action plan and interventions to protect health, ensure equity in decision-making, and restore the NEC community. This study will have local and circumpolar arctic implications for Indigenous communities. Locally, we will provide data and implement actions necessary for re- establishing the community at NEC. Given that thousands of such Cold War military sites exist throughout the Arctic, often in close proximity to Indigenous communities, our project may serve as a model for environmental and health monitoring and policy action by other Arctic Indigenous Peoples.