University Of Arizona

NIH Research Projects · FY 2025 · 2023-01

In 2020, more than 38.5 million (10.5%) U.S. households were without reliable access to sufficient quantities of affordable, nutritious food due to limited money or other resources. Food insecurity, poor nutrition, and economic disadvantage are critical social determinants of health that contribute to disparities in type 2 diabetes mellitus (T2DM), a serious diet-sensitive chronic disease affecting more than 20% of food insecure adults. Coping strategies favor inexpensive, ultra-processed foods that substantially diminish diet quality and are associated with increased incidence and severity of T2DM, including poorer glycemic control and excess body weight. Given that numerous social factors and systems contribute to and perpetuate food insecurity and poor diabetes outcomes, there is a demonstrable need for multilevel (individual, household, community) food-focused interventions to effectively and sustainably address the diet quality of persons with, or at risk of, T2DM. To our knowledge, no studies have rigorously tested whether intervention programs explicitly designed to improve both food and nutrition security of low-income persons with T2DM are feasibly delivered by personnel at a Federally Qualified Health Center (FQHC), acceptable to patients seeking care in their medical home, or are capable of producing clinically relevant changes in T2DM endpoints. These are questions we will explore in our proposed randomized wait-list controlled pilot study— Food and Resources Expanded to Support Health and Type 2 Diabetes (FRESH-T2DM)— in which our previously developed food and diabetes self-management education (DSME) intervention (FRESH) will be delivered to 50 adult FQHC patients with T2DM and food insecurity twice monthly, for 6 months. The FRESH intervention consists of bimonthly food provision; a series of recipes that feature FRESH foods; diabetes self-management education (DSME) resources; and four, 30-minute visits with an FQHC Registered Dietitian Nutritionist and Certified Diabetes Educator who will help participants utilize FRESH resources to meet personalized treatment goals. We will build upon our prior work and existing collaborations with colleagues at El Rio Community Health Center, a Federally Qualified Health Center serving >110,000 underinsured, uninsured patients and the Community Food Bank of Southern Arizona, a regional food bank serving 180,000 Arizonans across 5 counties to: (Specific Aim 1) Assess the feasibility, acceptability, and participant uptake of our FRESH intervention, delivered to 50 food insecure adults with T2DM at their medical home, El Rio Community Health Center, and (Specific Aim 2) Explore changes in blood glucose control, diet quality, food security, diabetes self-care behaviors, and health-related quality of life among participants at 3 and 6 months. Completion of our pilot study —submitted in response to NIDDK PAS-20-160 —will produce data to inform the rationale and design of a future definitive randomized controlled clinical trial, including recruitment, retention, adherence, and cost data. Our long-term goal is to produce a tested, efficacious model of coordinated care capable of replication and scaling across other FQHCs and food bank networks.

NIH Research Projects · FY 2025 · 2023-01

Project Summary/Abstract Alzheimer's disease (AD) is a fatal neurodegenerative disease affecting 5.5 million Americans. Despite many decades of research there is still no known cure. AD is a protein misfolding disease, where the Alzheimer's protein, Aβ, aggregates from a random coil entity into fibrils, which are highly organized aggregates containing a cross-β sheet structure. However, the nature of the toxic species in Alzheimer's disease remains unknown. More and more attention has been given to the possibility that Aβ aggregates within mitochondria, rather than extracellular deposits of Aβ, may be responsible for the onset and progression of the disease. Nature has developed mechanisms to prevent disease-associated protein aggregation, e.g. by the introduction of heat shock proteins (Hsp's), which are overexpressed when cells undergo stress. The most important Hsp's in mitochondria are Hsp60 and Hsp70, whereas Hsp60 is the only essential chaperone in bacteria, yeast, and mammals. It is known that Hsp60 is cytoprotective against many stressors in cells and is proposed to be directly protective against AD. However nothing is known about the mechanism of how this is achieved. Since almost nothing is known about the mitochondrial Hsp60 system, including its co-chaperone Hsp10, I will use aims 1 and 2 of this proposal to explore this chaperone in absence of substrate protein. I seek to solve the structure of different states of the chaperone during its protein folding cycle by cryo-electron microscopy (cryo-EM). Further, using novel solution-state NMR methods to study sparsely-populated states, I will investigate how the co-chaperone is involved in the reaction cycle. This will provide important information on the role of Hsp10 in substrate encapsulation. Further, during my independent phase I will use the fundament knowledge obtained through aims 1 and 2 to move on to aim 3, where I will study the interaction of Hsp60 with Aβ. I will not only elucidate the structure of long-lived major states of Aβ bound to Hsp60 after substrate encapsulation by cryo-EM, but also study the transient interaction of Aβ with Hsp60 during the initial phase of the substrate folding cycle, prior to encapsulation, by solution-state NMR. These studies will provide information about which Aβ species (monomer, oligomer, or fibril) interact with Hsp60 and elucidate their structural features. Aims 1 and 2 will provide insights into fundamental questions about the mechanisms chaperones use to efficiently fold proteins into their functional forms. Further the results from aim 3 will unveil details about how Hsp60 inhibits Aβ aggregation and prevents neurodegenerative diseases, and may open up novel therapeutic strategies against Alzheimer's disease.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY Headache affects more than 39 million Americans, yet a full understanding of the cellular and molecular pathology underlying headache has not been achieved; the discovery of novel, clinically useful therapeutics is therefore limited. This proposal will use complimentary techniques to 1) understand the mechanistic contributions of endocannabinoid system (ECBS) dysregulation to headache pathology and 2) validate a therapeutic strategy that enhances eCB tone to offer an improved option over current therapeutics. Studies will use two rat models of headache, cortical spreading depression (CSD) and medication-overuse (MOH) in male and female rats to define the cellular and molecular role played by the ECBS in headache. Recent clinical observations support the idea of Clinical Endocannabinoid Deficiency (CED) as a potential mechanism of migraine in some patients; however, studies providing evidence for a mechanistic role of eCBs in migraine are limited. Preliminary data suggest that both cortical KCl and medication overuse deplete 2AG and increase inflammation in the PAG, a midbrain region implicated in descending pain modulation. Further, headache symptomology could be induced by pharmacological depletion of 2AG in more female than male rats further supporting loss of eCB tone in headache in a sex-selective manner. These exciting findings led to the hypothesis that headache pain results from sex-dependent enhanced degradation of 2AG in the PAG by ABHD6 and MAGL leading to increased inflammation and loss of descending inhibition that drive pain behaviors. Three Aims will define the role of ABHD6 as the “gatekeeper” of 2AG availability for retrograde release (Aim1); determine the role of MAGL in terminating 2-AG actions during headache which may contribute to maintenance (Aim 2); and establish how ECBS dysregulation within the PAG occurs in males and females at the molecular and cellular levels during induction, maintenance, and recovery from headache like pain (Aim 3). Integration of these results between aims will clearly delineate the role of 2-AG within the PAG plays a role in headache induction and maintenance and validate increasing eCB tone by targeting either MAGL and/or ABHD6 as unique new targets for migraine therapy. Successful completion of this project will provide foundational rationale to initiate a drug discovery program selectively targeting the ECBS for migraine intervention to provide a better clinical option against headache as compared to current therapeutics.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY/ABSTRACT Tongue muscles, which are innervated by hypoglossal motoneurons (XIIMNs), are critical for survival given their role in suckling, swallowing, mastication, breathing and more advanced functions such as human speech. The hypoglossal motor nucleus is a bilateral collection of seven separate motoneuron pools, with motoneurons in each pool innervating one of the seven different tongue muscles. We recently showed that XIIMNs innervating the superior longitudinalis and genioglossus tongue muscles of neonatal rats have significantly different resting membrane potentials, action potential firing thresholds, and f-I curves, i.e., the change in firing rate as a function of injected current. These findings raise three very important questions: 1) what is the extent and nature of phenotypic diversity both within and between individual XIIMN pools? 2) what are the anatomic and ionic mechanisms that underlie this phenotypic diversity? 3) do structural and functional differences among XIIMNs in each pool map to unique gene expression profiles? We propose a rational and robust approach to address these questions: specifically, to describe the morphology, intrinsic membrane properties, and the transcriptome of muscle specific XIIMNs. Our initial targets are XIIMNs innervating the genioglossus, hyoglossus and superior longitudinalis muscles, as each muscle has different effects on tongue movement. Muscle-specific XIIMNs will be identified by injecting each of the muscles with a retrograde tracer conjugated to a fluorescent reporter. All experiments use brain tissue from neonatal rats 5-12 days of age. Key techniques include neuroanatomic tracing to define neuron morphology, immunohistochemistry, whole cell patch clamp electrophysiology and next- generation RNA sequencing. These basic science studies will identify unique molecular targets associated with functional and/or structural differences between the motoneuron pools. Without this fundamental information, interventions aimed at stimulating or inhibiting the activity of specific tongue muscles will be imprecise and may result in unintended outcomes. In contrast, specific knowledge of unique molecular targets will focus the development of therapeutic approaches aimed at stimulation and/or inhibition of specific tongue muscles. Preliminary data show several pool-specific differences in motoneuron function and gene expression, strongly suggesting that the proposed work will provide truly novel data on the anatomic, physiologic, and molecular underpinnings of phenotypic diversity within and between muscle-specific hypoglossal motoneuron pools. This, in turn, will lead to a major leap in our understanding of how the tongue muscles perform complex, coordinated behaviors such as suckling, swallowing, and defense of the upper airway during sleep, and will lay the foundation for the development of therapies aimed at controlling the activity of specific tongue muscles.

NIH Research Projects · FY 2025 · 2022-09

Summary Migraine is a prevalent neurological disorder affecting millions of people worldwide. The underlying pathophysiology of migraine likely involves diverse mechanisms within the trigeminal pain pathways and pain- related structures in the brain. Recently introduced medications targeting CGRP mechanisms including antibodies and small molecule CGRP receptor antagonists appear to act outside of the blood brain barrier. These medications are effective as preventive treatment in some patients implicating the role of meningeal CGRP in migraine pathology. However, many patients do not respond to these therapies and even those that do often experience “breakthrough migraines” suggesting the existence of non-CGRP peripheral migraine mechanisms. The brain circuits mediating migraine pain remain understudied. Cephalic nociceptive pathways involve activation of trigeminal afferents and transmission of the nociceptive signal through the second order cells in the trigeminocervical complex (TCC) to multiple brain regions. Human neuroimaging studies during a migraine attack have demonstrated activations in the amygdala and dorsal pons, encompassing the parabrachial nucleus (PBN). The PBN receives both exteroceptive and interoceptive sensory inputs and projects to multiple sites including the central nucleus of the amygdala (CeA), an area mediating emotional aspects of pain. Our preclinical studies using pharmacological and optogenetic activation of dural afferents, neuronal tracing and behavior suggest a functional pathway from the dura mater to TCC, PBN and CeA that may promote migraine-like pain. In this proposal, we will use opto/chemo-genetic methods, microscopy with immunostaining and RNAscope, CRISPR-Cas9 (i.e., CRISPR) genetic manipulations, electrophysiology, calcium imaging and pain behavior to investigate if, and how, the PBN→CeA pathway may promote migraine-like pain elicited by activation of dural afferents using several different approaches in male and female mice. Aim 1 will measure the effects on markers of neural activation in brain networks (microscopy) and consequences on pain behavior; Aim 2 will use brain slice electrophysiology and calcium imaging with pharmacological, CRISPR or opto/chemo-genetic manipulations of PBN outputs to evaluate plasticity of CeA cell types; Aim 3 will use chemogenetic inhibition of PBN outputs or CRISPR deletions in the CeA to evaluate possible inhibition of migraine pain behaviors. Our studies aim to address significant gaps in our knowledge of central pathways of migraine pain. We will determine the potential relevance of the central PBN→CeA circuit as a common pathway of migraine pain that can be engaged by CGRP-dependent and CGRP-independent peripheral mechanisms.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Effective medical nutrition therapy (MNT) is needed to stem the alarming increase in Type 2 diabetes (T2D) related morbidity and mortality in Benin, a French sub-Saharan African (SSA) country, where diabetes prevalence has quadrupled from 3% to 12.4% in the last decade, reaching 22% in some areas. The impact of the disease reaches beyond those living with the condition to their families, society, and the healthcare system, warranting strategic efforts to reduce its burden. Following a diet consistent with MNT guidelines is well- documented to help people with T2D achieve better glycemic control, reduce cardiovascular risks and prevent complications. However, despite these benefits, low adherence to dietary recommendations is consistently reported. The reasons include lack of nutrition knowledge, difficulty translating recommendations to practice, cultural acceptability of the diet, and access to healthy foods. Thus, culturally appropriate dietary recommendations based on locally available and accepted foods through guided counseling are key to improving dietary adherence rates among T2D patients in the country. The purpose of this International Research Scientist Development Award (K01) is to empower Dr. Halimatou Alaofè to develop a research program that integrates innovative approaches to nutrition care delivery to optimize diabetes control in low- income African populations. To achieve this goal, mentorship and training in this award are proposed to support: 1) additional training in menu planning and recipe development, social and behavioral sciences, and research methodologies; 2) protected time to conduct the planned patient-oriented research; and 3) the collection and publication of preliminary data to facilitate the next steps in the research program. Under the mentorship of Drs. John Ehiri and Waliou Amoussa Hounkpatin, as well as co-mentors and collaborators from the US and Benin, Dr. Alaofè plans to 1) adapt an existing MNT intervention that combines a 4-week local food menu plan based on the 4A framework (adequacy, acceptability, accessibility, and availability of foods) with individual counseling to meet the Benin nutrition therapy guidelines for T2D patients; 2) develop SmartMenu, a culturally tailored MNT for T2D patients using codesign methods; and 3) assess the feasibility, acceptability, and preliminary efficacy of SmartMenu for Benin patients with T2D. The anticipated outcome of this study is high-quality pilot data that will serve as the foundation for an R01 application to conduct a larger pragmatic randomized controlled trial to assess the intervention’s cost-effectiveness and sustainability. We hypothesize that a culturally tailored MNT combining individual counseling and menu plans that address dietary adherence barriers through the 4A framework will facilitate patient acceptance and adherence to MNT guidelines. If successful, this model could significantly improve nutrition care in resource-limited settings. The training and research activities delineated complement the candidate’s background in nutrition education and diet modification and will position her to contribute substantially to nutrition care delivery system change.

NIH Research Projects · FY 2025 · 2022-09

Engineering of bacterial synthetic multicellular systems and materials hold promise for many health- relevant applications such as modular drug biosynthesis, living diagnostic devices, and synthetic biofilm research models. To date, bacterial synthetic biology has largely focused on the scales of molecules and single cells. Equivalent work on bacterial synthetic consortia is much less advanced, in significant part due to the previous lack of suitable synthetic and genetically encoded cell-cell adhesion tools to control the assembly, development, and functionality of multicellular systems. We recently developed the first such synthetic cell-cell adhesion toolbox, as well as tools for optogenetically controlling cell-surface deposition and patterning. The specific objectives of this research are to significantly advance these synthetic cell-adhesion tools, and to develop design principles and predictive modeling tools that enable consortia engineering and patterning that integrate all relevant length scales (i.e., molecular, cellular, and multicellular), and ultimately pave the way for medially relevant applications. Our main hypothesis is that we can significantly advance our control over the strength, specificity, and subcellular localization of synthetic adhesion proteins in Escherichia coli, which will allow rational tuning of consortium-level biophysical properties such as porosity and viscoelasticity, and which will ultimately enable versatile multicellular consortium engineering and patterning. This work will constitute a foundation for various biomedical applications such as biocompatible materials, multicellular plug-and-play pathway engineering, targeted in-vivo drug delivery, and living diagnostic devices. Our interdisciplinary methodology combines synthetic biology, biophysics, instrumentation and modeling. All experiments will be done in a quantitative manner. The proposed investigations include three independent yet synergistic Specific Aims motivated by our hypothesis: (Aim 1) Advance the functionality of the synthetic adhesin toolkit at the subcellular level; (Aim 2) Achieve engineering control over synthetic consortium properties such as viscoelasticity and porosity at the scale of 10-100 µm; and (Aim 3) Achieve higher-level consortium patterning on the scale of centimeters and demonstrate potential for medical applications. The PI (Prof. Riedel-Kruse) and his team are well-suited for this project as we have significant expertise in synthetic biology, biophysics, instrumentation (e.g., microfluidics, imaging), and modeling genetic circuits and biophysical systems across scales. We developed the first synthetic cell-cell and optogenetic cell-surface adhesion toolboxes in bacteria. Multiple collaborators provide additional domain expertise in key areas. Overall, this project's innovation lies in establishing synthetic adhesins as an essential and integral component of the synthetic circuit-engineering toolbox and in establishing a novel paradigm for modular engineering of multicellular living materials. Accordingly, this project will broadly impact the engineering of synthetic consortia for basic research as well as enable a dynamic spectrum of future applications in health.

NIH Research Projects · FY 2026 · 2022-09

ABSTRACT Use of cancer screening services for Latinx populations has lagged far behind the use for non-Latinx White populations; this has led to later stage detection and worse survival for Latinx people. Clinic-delivered video- texting combined with neighborhood-level navigation services is a telehealth-community solution that offers great promise for addressing cancer screening disparities, as it reduces multi-level barriers to care. Our proposed project, Community Partnership for Telehealth Solutions to Counter Misinformation and Achieve Equity (PRIME), will leverage partnerships among research centers, clinics, and community organizations to provide personalized support services informed by local knowledge and neighborhood-level data, and apply and evaluate novel technical and communication solutions. The goal of PRIME will be to improve colorectal cancer-related care delivery and reduce disparities in telehealth access and care by developing and testing telehealth tools and technologies using novel research methods. This research will use rapid methods and adaptive-evaluation processes and will apply the Health Equity Implementation Framework and components of the Reach Effectiveness-Adoption Implementation Maintenance (RE-AIM) framework, with an emphasis on effects on disparities in implementation and evaluation. PRIME will assess neighborhood-level social determinants of health and adapt and optimize a telehealth-solution to improve the quality, reach, and effectiveness of colorectal cancer screening in predominantly Latinx clinic populations aged 45-54 (Aim 1); test technical and communication solutions that use video-text messages to educate patients about colorectal cancer screening, link them to community-based services, and provide personalized navigation to support to improve uptake of screening and follow-up care, using a pragmatic stepped-wedge design (12 neighborhoods; 3,000 patients aged 45-54; Aim 2); and assess multi-level moderators of program effectiveness (including neighborhood-level social determinants of health, ethnicity, and preferred language) and organizational-level barriers and facilitators to implementation; and scale-up the program across neighborhoods and partnering clinical practice sites (n = 4; Aim 3). This project will leverage local resources and strengthen connections among health systems and trusted community-based organizations, increasing the likelihood of program sustainment. To ensure the project’s success, we have assembled an engaged group of community partners, including payers, health centers, community-based organizations, and patients, along with a multi-disciplinary research team of nationally renowned clinicians and scientists. If successful, the program will result in ready-to- implement strategies to reduce cancer-related disparities and improve preventive cancer care access, quality, and health outcomes for Latinx populations.

NIH Research Projects · FY 2024 · 2022-09

PROJECT ABSTRACT The FMR1 premutation (PM) affects ~1 in 150 women and ~1 in 470 men in the United States, and can have a significant effect on physical and mental health. Forty percent of males and 16% of female PM carriers are diagnosed with fragile X-associated tremor/ataxia syndrome (FXTAS) between 55-60 years of age, though who will manifest the disease is unknown. FXTAS is characterized by executive dysfunction, gait ataxia, and intention tremor. In the absence of clear markers of disease progression among PM carriers, it is essential to characterize language. Language predicts neurocognitive decline in related conditions (e.g., Alzheimer’s) 10-20 years prior to diagnosis. Language is a promising candidate marker of FXTAS as female PM carriers show age-related changes in pragmatics and lexical-semantics, which have been linked to executive dysfunction. Executive function and language difficulties may adversely impact quality of life. No studies to date have examined language among both male and female PM carriers, limiting our understanding of potential preclinical markers of FXTAS. As a first step, the proposed project will comprehensively examine language among PM carriers in comparison to healthy controls. Without such data, this severely limits our ability to (a) fully understand the impact of the FMR1 PM, (b) examine cognitive correlates implicated in FXTAS (i.e., executive dysfunction), and (c) understand the implications of language use and executive dysfunction on quality of life. This proposal addresses these limitations with three aims: 1) Examine pragmatic and lexical-semantic language among male and female PM carriers in comparison to healthy age- and sex- matched controls; 2) evaluate interactions of age and executive functioning on language; and 3) assess interactions between language and executive function on quality of life. This study will be completed virtually with a cross-sectional sample of 60 PM carriers (30 males, 30 females) without FXTAS and 40 healthy age- and sex-matched controls (20 males, 20 females) between the ages of 35-55. Participants from across the U.S. will be recruited. Participants will complete virtual language elicitation and executive function tasks, and self-report measures of executive function and quality of life. Results from this proposal are expected to inform the nature of language differences among PM carriers compared to controls. It is also expected that age and executive function will interact to adversely influence pragmatic and lexical-semantic language in PM carriers but not controls, which would implicate potential neurocognitive decline. Finally, we anticipate that poorer language and executive functions will adversely affect quality of life among PM carriers. This proposal is consistent with the mission of the National Institutes of Health to enhance health and reduce disability by expanding our understanding of a neurodegenerative condition associated with a common genetic variant, the FMR1 PM. It also contributes to the strategic mission of the National Institute on Deafness and Other Communication Disorders by informing knowledge on the basis of language impairments among individuals with neurodegenerative disorders and links with quality of life.

NIH Research Projects · FY 2025 · 2022-09

The discipline of biomedical informatics is primed to foster scientific innovation and promote health for all. To truly harness the potential of multitude of methods, theories, and application in the field of informatics, there is pressing need to enrich the research community by expanding participation from a range of relevant disciplinary perspectives. Recent educational research suggests that intentional connections to place, community, and local needs may be critical to welcoming and retaining individuals in biomedical research. Place-based education, a form of community-based learning, aims for purposeful student engagement that is responsive to local societal and health needs, including its history, people, and ecology. Such learning cultivates critical and analytical skills and actively engages students in real-world problem solving and holds much potential for increasing student engagement in scientific research as well as in interdisciplinary areas such as informatics. In this project, we propose to develop a truly interdisciplinary, Place-based Health Informatics Research Education (PHIRE; pronounced as “fire”) program that brings together expertise from data science, systems science and engineering, health informatics, and public health and infuses regional assets and needs from the community into the training and curricula. The vision of the PHIRE training program is to recruit and prepare a talented pool of 50 undergraduate students (10 students per year) to conduct scientific research in health informatics with an inherent focus on addressing biomedical and public health challenges. The PHIRE training program includes three major components: (1) a 12-week summer research experience, (2) a thematic co-curricular training in health informatics, and (3) preparation for graduate school or other research careers in health informatics.

NIH Research Projects · FY 2025 · 2022-09

This R35 application describes our continuing and expanding program to develop and apply computational methods to study how protein dynamics on many timescales contributes to enzymatic catalysis, and how some enzymes are crafted by evolution to make use of protein dynamics on multiple timescales. This knowledge will eventually inform approaches to design artificial enzymes – a grand challenge, as yet unmet. Our studies of enzymatic catalysis began years ago with the first application of Transition Path Sampling to chemical reaction in enzymes. The generation of reactive trajectory ensembles along with reaction coordinate identification allowed us to postulate the concept of the protein promoting vibration: rapid protein dynamics at or near the active site that are directly coupled to passage over the transition state barrier to reaction. Such motions were found in multiple enzyme systems (but not all,) and their importance was verified by experimental collaborators. More recently, application of these methods to artificial enzymes that are subjected to optimization by laboratory evolution has shown the evolutionary process introduces such motion into a static design and we have identified protein structural changes that allows the creation and coupling of the dynamics. “Theozymes” created by de novo static structural methods have had limited success, while laboratory evolution has allowed these proteins to develop significant catalytic power, We will continue and significantly expand our program on this challenging topic through extensions of both methodologies employed and with application to both natural and laboratory evolved enzyme families. In addition to understanding how protein evolution leads to the coupling of rapid dynamics to barrier passage, we will extend our approaches to be able to map how motions far more remote in time from the passage over the chemical barrier can also be central to function. We will also develop methods that demonstrate how rapid motions can prime the system for millisecond conformational change. Our goals for the program are to understand how protein dynamics ranging from sub picosecond promoting vibrations to microsecond domain motions to millisecond conformational motion are potentially inter-related and help form enzyme function, and how such motions are orchestrated by the structure crafted by evolution. The development of such tools coupled with studies of both laboratory and naturally evolved enzyme families will allow the isolation of a “dynamics toolbox” employed by selection. We have already found how in one laboratory evolved enzyme the introduction and loss of hydrogen bonds in strategic locations results in the creation of a promoting vibration; successful completion of the proposed program will expand such investigation to the full range of protein motion timescales appropriate to catalytic turnover, along with the architectural changes needed to create such dynamics in the protein catalyst.

NIH Research Projects · FY 2026 · 2022-09

Project Summary Pressure ulcers affect 2.5 million people in the United States and cost the healthcare system an estimated 11.6 billion dollars annually. A major percentage of affected individuals are over sixty-five years of age and this number is growing rapidly in the United States. A greater incidence of immobility, vascular disease and diabetes is commonly seen in aged populations and places them at an increased risk for developing pressure ulcers. Thus, there is a pressing need to develop new therapies that will effectively treat pressure ulcers in the aged. With prior NIA support, we have found that aging is characterized by the progressive loss of critical molecular and cellular pathways responsible for normal wound healing, specifically hypoxia-inducible factor 1-alpha (HIF- 1α). We have also identified that these changes can be reversed using an FDA-approved small molecule drug, deferoxamine (DFO). Building on our preliminary data, we will first define the aberrations of HIF-1α in healthy, unwounded skin from young and aged patients (Specific Aim 1). We will then confirm the HIF-1α signaling dysfunction in pressure ulcers using biopsy specimens from both young and aged patients. (Specific Aim 2). To define the alterations in hypoxia signaling within the skin cells of young versus aged patients, we will subject the collected tissue for protein analysis and single cell sequencing. We will then determine the optimal delivery system for DFO through both intact stratum corneum as required for treating pressure ulcers in a porcine model (Specific Aim 3). This large animal model is selected since porcine skin has significant similarities to the human skin. Our results will validate the effectiveness of our drug delivery system and provide a novel strategy to deliver other hydrophilic small molecule drugs across intact human skin. Taken together, this pre-clinical study will lay the groundwork for a pilot human trial targeted at treating pressure ulcers with the transdermal DFO delivery system, which will for the first time provide a pharmacologic therapy to treat pressure ulcers in aged patients.

NIH Research Projects · FY 2025 · 2022-09

Satisfaction of Search (SOS), where an abnormality is missed after a previous abnormality was found, is a pervasive problem across different types of images, yet their impact on breast cancer detection is un- known. Radiology and cognitive science have independently investigated why, how, and when SOS oc- curs. The objectives of this grant are to bridge cognitive science research techniques with radiology to de- termine: 1) the neural underpinnings of SOS, 2) the extent to which SOS affects breast cancer detection, 3) the extent to which advancements in imaging technology alleviates SOS, and 4) how expertise affects SOS error rates. My central hypothesis is SOS contributes to breast cancer misses because a found ab- normality becomes an attentional template. An attentional template is known in cognitive science as an enhanced representation of a target maintained within memory and biases attention in search. The ra- tionale for this project is when a first target is detected, it becomes an attentional template that utilizes and biases attention necessary for detecting subsequent targets making searchers prone to SOS. However, SOS may be mitigated: 1) by changing search strategies to counteract a first target becoming an atten- tional template, 2) with advances in imaging technology, and 3) with expertise in breast cancer detection. These influences need to be investigated to determine the extent to which they can alleviate SOS. This work will pursue four specific aims to test the central hypothesis: 1) identify a mechanistic account for SOS, 2) determine the extent to which SOS accounts for breast cancer misses, 3) compare SOS between mammography and tomosynthesis, and 4) compare SOS error rates between different levels of expertise. The research proposed is innovative because it will: 1) identify a mechanism responsible for SOS; 2) use EEG and eye-tracking to study the neural basis of SOS; 3) use realistic, virtual breast images to investi- gate the extent to which SOS affects breast cancer detection; 4) determine the extent to which the SOS results from simplified search displays replicate in realistic-looking medical images; 5) be the first time SOS is investigated with 3D imaging; and 6) determine the extent to which SOS rates vary by expertise in breast cancer detection. The proposed research is significant because: 1) it will develop a neural under- standing of how SOS contributes to cancer detection; 2) determine how SOS varies across different search modalities and by expertise; and 3) ultimately improve breast cancer detection by determining how these influences impact SOS rates. To complete the proposed research, in pursuit of his long-term goal of becoming an independent researcher at a Tier-I institution, the PI will train using EEG and eye-tracking. His training will include auditing graduate courses, attending workshops, weekly meetings/training with his mentors, and bi-annual assessments of his EEG/eye-tracking expertise with his mentors.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The Integrated Environmental Science and Health Risk Laboratory, in collaboration with multiple Arizona partners, propose to launch the project, “Building a culture of health in the green: Participatory learning and action to address air and soil quality in rural underserved communities”. Our overall goal is to create a national deep engagement learning model of Science, Technology, Engineering, Arts and Math (STEAM) education that empowers students as environmental health advisory board members, photographers, scientists, and designers living in environmental justice (EJ) communities. EJ communities refers to areas affected by disproportionate exposure to environmental hazards and increased vulnerability to those hazards. There are fundamental and critical challenges that exist in environmental justice communities. Youth growing up in these spaces need the preparation to navigate through these challenges and be the future pillars of structural change. Students will acquire the tools, skills and support needed to (1) interpret intersectionality and environmental data and (2) communicate project results and recommend actions at local, regional, and national levels. The proposed project centers on the social and environmental factors that affect health and justice for 8th - 12th graders in rural Arizona communities with significant environmental health hazards posed from historical or active resource extraction activities. Intersectionality and environmental health and data literacy are essential for deriving meaning from the results of scientific inquiry. Potential outcomes for the proposed project are increased understanding and integration of intersectionality, environmental health (specifically air and soil quality), and data literacy among both youth and researchers, and the application such information to achieve social justice and health equity in EJ communities. This research education program aims to leverage intersectionality’s transformational power and help move the collaborating EJ communities closer to achieving social justice and health equity. Through the approach and rigorous learning research, the outcomes of this work will provide the evidence-base needed for building a culture of health in the green.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Stroke is the fifth leading cause of death in the United States. Approved stroke therapies are limited by narrow treatment windows, the risk of hemorrhagic transformation, and reperfusion injury. Therefore, there is a critical need for neuroprotective drugs that can improve post-stroke neurological performance. Currently, 3-hydroxy-3- methylglutaryl coenzyme A (HMG CoA) reductase inhibitors (i.e., statins) are given to stroke patients due to their proven utility in improving cognitive and motor outcomes. Studies in our laboratory have uncovered a specific biological mechanism that enables statins to be effective drugs for stroke treatment: transport across the blood- brain barrier (BBB) via the endogenous uptake transporter organic anion transporting polypeptide 1a4 (Oatp1a4). We have shown, for the first time, that Oatp-mediated transport is required for atorvastatin to reduce cerebral infarction volume and improve sensorimotor performance at 24 h following transient middle cerebral artery occlusion (tMCAO). We also observed increased atorvastatin uptake in female rats subjected to tMCAO as compared with their age-matched male counterparts; however, it is unknown if these differences in Oatp- mediated transport at the BBB cause variations in atorvastatin’s ability to prevent stroke progression and/or worsening of neurocognitive deficits in the acute/subacute period. Our goals are to assess the role of sex as a biological variable on statin transport in the ischemic brain and to determine how these differences affect statin efficacy as stroke therapeutics. The central hypotheses of this F31 application are i) that functional expression of Oatp1a4 at the BBB is different in males as compared to females following tMCAO; and ii) that statin neuroprotective properties and/or their effects on post-stroke neurological outcomes are influenced by sex-dependent differences in BBB Oatp1a4 activity. Two aims will test these hypotheses: Aim 1: Investigate sex-dependent differences in Oatp1a4-mediated transport of statins at the BBB in stroke. We will perform our studies in male and female SD rats using the tMCAO model. Oatp1a4 localization and protein expression will be assessed using confocal microscopy and western blotting, respectively. Blood-to- brain transport of statins will be measured using in situ brain perfusion, a state-of-the-art methodology to study drug transport at the BBB. Aim 2: Evaluate sex-dependent differences in statin-associated neuroprotection and functional neurological recovery in stroke. In tMCAO operated male and female SD rats, we will use confocal microscopy and western blot analysis to examine molecular biomarkers associated with neuroprotection. We will also assess motor and cognitive performance in tMCAO-animals using robust behavioral tests (i.e., rotarod analysis, Morris Water Maze, Novel Object Recognition test). Overall, this project will provide critical mechanistic data on efficacy of statins as neuroprotective drugs for stroke. Furthermore, this project directly aligns with NIH goals in studying sex as a biological variable in stroke.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT The goal of the work outline in this proposal is to understand the fundamental signaling that controls cell fate to maintain tissue homeostasis. Epithelial tissues demonstrate an intrinsic ability for their constituent cells to organize and maintain a steady-state of form and function. Many disease states lack these intrinsic controls. Epithelial tissues of the human body are in a constant state of renewal. Our understanding of the signaling systems that control how these complex epithelial tissues maintain robust organization is incomplete. Essential for progress, we need a quantitative understanding of signaling at the single-cell level in the context of physiological conditions to reveal systems-level behaviors that can be targeted therapeutically. We focus on protein kinases as critical mediators of signaling in the cell, which are well known to play prominent roles in tissue function and drivers of disease. Our kinome-wide studies have identified GSK3 and CLK3 as major tissue homeostasis regulators that govern the balance between proliferation and differentiation. In Project 1, we hypothesize GSK3 requires multiple suppressive inputs that uniquely produce different fate outcomes ranging from stem cells, transit-amplifying, and differentiated. We will provide the first systems-level mapping for multiple inputs onto GSK3 dynamics and how these dynamics are decoded into distinct cellular outcomes of renewing epithelium. In Project 2, we hypothesize CLK3 is a gatekeeper controlling stem cell fate through transcriptome regulation. We will define the activity of CLK3 in the stem cell niche as a regulator of expression and splicing of Wnt-target genes to promote stemness. Our approach uses high-throughput quantitative microscopy to measure single-cell behaviors in physiological organoid homeostatic culture models. Our research will define the regulatory mechanisms for two critical kinases, GSK3 and CLK3, and discover novel signaling circuitry needed for the accurate organization of renewing and regenerative epithelia, uncovering new strategies for treating diseases of regenerative tissues.

NIH Research Projects · FY 2026 · 2022-08

PROJECT ABSTRACT: Adversity is a ubiquitous human experience associated with increased risk for poor health outcomes, including Alzheimer’s disease (and related dementias; AD/ADRD) and accelerated biological aging. Although the research linking adversity to cognitive functioning and aging is robust and growing, much of the current evidence is correlational, leaving questions of causation largely untested. Behavior genetic methods provide powerful tools to evaluate putatively causal associations by accounting for genetic and shared environmental confounds. In this competitive revision application, we expand the scope and scientific impact of an ongoing NIH-funded R01 study (#AG078361-01) with a new set of aims studying adversity, cognition and biological aging using behavior genetic methods. Data collection in the parent project—which includes a large sample of twins from the Washington State Twin Registry (WSTR; final target N = 1,782; 891 twin pairs)—is ongoing and the goals of this project are to leverage existing data in the WSTR to address the following aims: Aim 1a-1b: Using cotwin control design modeling, test whether the associations between adversity and both cognitive functioning and biological aging are consistent with causal associations. After accounting for shared genetics and environmental influences, we hypothesize that twins who experience greater trauma, stress, and adversity will also exhibit poorer cognitive function and more rapid biological aging. Aim 2a-2b: Test whether adversity is associated with unhealthy behaviors, as well as whether unhealthy behaviors are associated with poorer cognition and accelerated aging, consistent with a causal association. We hypothesize that twins with more adversity within twin pairs will have more unhealthy behaviors, which will be associated with poorer cognitive functioning and faster biological aging. Exploratory Aim 2c: Test which health behaviors explain associations between adversity, cognition functioning, and biological aging. We will achieve these aims by combining WSTR assessments and data collection in the parent grant with support from new investigators with extensive expertise in the study of trauma, stress, and adversity. The associations between adversity and accelerated cognitive and biological aging are of significant public health importance, but the field needs to move beyond correlational research alone. When realized, the aims of this project will allow us to test whether a range of adverse experiences contribute to future health in a manner that may be consistent with a causal influence. This work will provide key evidence for future interventions that might slow biological aging and delay the onset of AD/ADRD.

NIH Research Projects · FY 2026 · 2022-08

While the earliest phase of Alzheimer’s disease (AD) pathology is often described as “clinically silent”, prior work raises the possibility that early AD is associated with detectable alterations in autobiographical thought – a class of cognition encompassing memories, plans, and other mental simulations related to our personal lives. Here we introduce two multi-modal studies that investigate whether cognitive markers of early AD neuropathology can be detected by deploying smartphone applications (apps) to track autobiographical thoughts in everyday life. Using two smartphone apps developed by our team to naturalistically assess cognition, the proposed studies will a) examine the sensitivity of real-world autobiographical thoughts to AD plasma and brain biomarkers in clinically normal older adults, b) reveal the predictive and scalable potential of measuring autobiographical thoughts in older adults for a host of longitudinal AD biomarker and associated health outcomes, and c) shed light on neurocognitive autobiographical thought characteristics that may separate normal from abnormal cognitive and brain aging. MPIs Dr. Grilli and Dr. Andrews-Hanna have formed a team of researchers with expertise in smartphone-based assessment of cognition, autobiographical thought, functional magnetic resonance imaging, healthy and pathological aging, and longitudinal analysis of large datasets. Leveraging our team’s interdisciplinary expertise, we will execute an innovative two-pronged project harnessing in-lab, at-home, and online assessment methods that will evaluate the relationships of AD biomarkers and aging to the autobiographical thoughts of 1,225+ adults, with a subset completing additional in-lab experimental cognitive tests, neuroimaging, plasma biomarker assays, and longitudinal follow-up. In Aim 1, we will test the hypothesis that among clinically normal older adults, smartphone measures of autobiographical thoughts are sensitive to plasma AD biomarkers, and resting state functional connectivity in the default network, a brain network targeted by early AD. Aim 2 tests the hypothesis that these smartphone measures predict future plasma biomarker accumulation among older adults who were clinically normal at enrollment, as well as future resting state functional connectivity of the default network, and daily psychosocial / instrumental decline. Aim 3 deploys one of our smartphone apps to a large remote, clinically normal, and genotyped cohort, providing an opportunity to evaluate questions about effects of age and genetic risk for AD on autobiographical thoughts at an unprecedented scale. Across the aims, we also examine how smartphone measures of autobiographical thoughts compare to in-lab cognitive tests, and if they improve sensitivity to aging and AD risk. To our knowledge, this project will be the first to use smartphones to track autobiographical thoughts as a means to identify cognitive correlates of AD biomarkers, despite theoretical tenets and evidence that doing so could tap into the primary brain pathway of AD. Ultimately, our mobile tools may lead to more accessible cognitive tests of early AD, including initial stages of amyloid and tau, with broad impact for scientists, clinicians and patients worldwide.

NIH Research Projects · FY 2025 · 2022-08

Abstract Knee osteoarthritis (KOA) is a major public health problem that affects ~30 million US adults, with increased morbidity and premature mortality. KOA has been identified as one of the biggest unmet medical needs because there are no FDA-approved treatments available to prevent, slow or halt OA progression. Development and evaluation of potential OA treatments has been hampered in part by KOA typically being a slowly progressive disease, though progression is variable and some knees undergo rapid structural or clinical progression. Feasible high-throughput methods are needed to gain insights as to how to identify rapid progressors. We have developed a rapid, reproducible, and responsive quantitative software tool, Quantitative Radiographic Software Scores (QROS), to measure minimal joint space width (JSW), fixed JSW (fJSW), variance of 14 fJSW locations; degrees of knee alignment/ malalignment; and orientation and roughness of trabecular bone texture (TBT). The overarching goal of our research is to identify knees at high risk of rapid progression, and our core concept is that quantitative assessment of a novel combination of baseline structural features on radiographs will identify a group of knees with rapid structural and/or clinical progression in KOA. Our central hypothesis is that abnormalities of structural features on knee radiographs will be determinants of varying combinations of worsening rates of structural and/or clinical progression in KOA. Our approach takes advantage of a unique opportunity to innovatively assess radiographic features on ~10,500 individuals by combining three of the largest and most racially diverse longitudinal observational studies of individuals with or at risk of developing KOA: the Johnston County Osteoarthritis Project (JoCoOA), the MulticenterOsteoarthritis Study (MOST), and the Osteoarthritis Initiative (OAI). All three cohorts have fixed flexion, weightbearing knee radiographs that were acquired with identical protocols over multiple visits, and comparable data on demographics, KOA risk factors, and WOMAC pain and function. Our specific aims are: 1). To quantitatively assess structural and clinical progression of KOA, separately and combined; 2). To identify the structural determinants of rapid KOA progression; and 3). To determine the knee- specific probability and time to knee replacement (KR) based on structural and clinical determinants.

NIH Research Projects · FY 2025 · 2022-08

SUMMARY Hearing impairment is a common and disabling sensory defect which in a subset of individuals can be due to an abnormal cochleovestibular anatomy. Cochleovestibular (CV) and cochleovestibular nerve (CVN) anomalies can significantly impact a child’s development and currently pose challenges in treatment and management. Little research has been done to understand the etiology of these malformations, especially those that are non- syndromic and severe, such as cochlear aplasia. There is a crucial need to better understand the underlying molecular mechanisms of these conditions to aid in diagnosis, intervention and management. In addition, health disparities exist in the molecular diagnosis and treatment of hearing impairment (HI) in Hispanics, as the molecular etiology of HI has been scarcely studied in this ethnic group. It is imperative to study the etiology of CV/CVN anomalies in diverse racial/ethnic populations to understand which genes/variants are a frequent cause of this disorder in each population. Molecular diagnostics and treatment can therefore be tailored based on population-specific information. We hypothesize that a significant subset of severe non-syndromic CV/CVN anomalies has a genetic etiology, which may differ between populations, and knowledge of this information will improve our understanding of inner ear development. Our preliminary research suggests that rare genetic variants, including de novo variants, are implicated in the development of severe CV/CVN anomalies. Our proposal leverages genomics data and temporal bone imaging data to unravel the molecular basis of non-syndromic CV/CVN malformations. To achieve this, we will 1) recruit and establish a large genomic database of racially/ethnically diverse families with CV/CVN malformations which have been phenotyped in detail. 2) Next, we will determine the genetic spectrum of underlying variation implicated in CV/CVN malformations in both Hispanic and non-Hispanic individuals. 3) Last, using recruited and existing cohorts of individuals with CV/CVN malformations and prelingual sensorineural hearing impairment, we will identify novel causal genes implicated in CV/CVN malformations and assess their expression during early craniofacial and inner ear development. We have assembled a team that has the collective expertise to achieve these aims as well as a prior track record of productive collaboration. This work will elucidate the genetic architecture of severe non-syndromic CV/CVN malformations diverse ethnic/racial populations and improve our basic knowledge of human inner ear development and the mechanisms leading to abnormal development. This knowledge can then be used to improve molecular diagnostics, guide therapeutic intervention and management, predict outcomes, and develop novel therapeutic approaches benefiting individuals of diverse ethnicity/racial background.

NIH Research Projects · FY 2026 · 2022-08

Abstract The intestinal epithelium is vital to maintain the barrier between the body and the outside world, and membrane trafficking is essential for both the development and maintenance of this barrier. Abnormal membrane trafficking can result compromised barrier leading to intestinal disease, including inflammatory bowel disease. Autophagy is a specialized membrane trafficking process that allows cells to respond to changes in metabolism. In addition, autophagy is important for maintenance of the epithelial barrier and the innate immune response, mediating the isolation and degradation of intracellular pathogens. Furthermore, LC3-associated phagocytosis (LAP) is important for uptake and degradation of pathogens, and dysfunction of this pathway is associated with hyperinflammation. Importantly, variants of proteins in the autophagic and LAP pathways have been linked to increased susceptibility to Inflammatory Bowel Disease (IBD), and particularly Crohn’s disease, although the molecular mechanisms that underlie this connection remain incompletely understood. MAMDC4 is an integral membrane protein that localizes to endosomes of the intestinal epithelium. Deletion of MAMDC4 compromises intestinal enterocyte morphology, and RNAseq studies have indicated that MAMDC4 is down- regulated in IBD. In previous work, we found that MAMDC4 interacts with the small GTPase Rab14. In professional phagocytes, Rab14 is recruited to phagosomes and prevents phagosome-lysosome fusion but its role in the intestinal epithelium is unknown. In our preliminary data, we show that Rab 14 is present on autophagosomes and both Rab14 and MAMDC4 are present on membranes containing invading bacteria. Furthermore, deletion of MAMDC4 results in accumulation of autophagy and lysosomal markers on tubular membranes. These results suggest that MAMDC4 and Rab14 act in a molecular network to maintain mucosal immunity through control of autophagy or LAP. In this proposal we will use intestinal epithelial cells in culture, organoid culture, and patient-derived organoids to define the role of autophagy and/or LAP in intestinal homeostasis.

NIH Research Projects · FY 2025 · 2022-08

Summary This proposal focuses on the structure and function of nebulin, an unusually large sarcomeric protein that is expressed in skeletal muscle. The giant size of nebulin has made it challenging to elucidate its functions but its importance is supported by the many nebulin mutations that cause nemaline myopathy (NEM2), the most common non-dystrophic congenital myopathy. The protein structure of nebulin consists of a large number of simple repeats that are actin-binding, most of which are organized into super-repeats (SRs). Approaches to treat NEM2 are sorely lacking and gaining an in-depth understanding of the many roles of nebulin in muscle structure and function is essential. We will comprehensively study nebulin, building on major advances that we and others have made in recent years. To help achieve our goals we utilize mouse models, some of which mimic severe and others milder NEM2, as powerful tools for our basic science and translational studies. We will investigate nebulin's functions from the single-molecule to the intact muscle levels, using multidisciplinary approaches that involve transcriptomics, proteomics, super-resolution imaging, low-angle X-ray diffraction, and biomechanics. Aim 1 focuses on thin filament length regulation. Our recent work supports that in slow muscle, nebulin collaborates with leiomodin-2 (Lmod2), with nebulin regulating the length of a proximal thin filament segment and Lmod2 regulating the length of a distal segment that is nebulin-free. Here we will critically test this dual length regulation model and study whether it has translational potential, by determining whether upregulating Lmod2 is an effective treatment for severe nebulin-based nemaline myopathy. Aim 2 studies the functional significance of weak actin-binding of centrally-located nebulin SRs that bind actin more weakly than those near the ends of the molecule. This is likely functionally important, considering that a mutation that increases the binding affinity of a central SR causes a skeletal muscle myopathy in patients world-wide. We will study mouse models in which centrally located weak-binding SRs have been converted into strong-binding SRs using mechanical assays and X-ray diffraction on intact muscle. Aim 3 studies the C-terminus of nebulin, its layout in the Z-disk, and the functions of nebulin's differentially expressed Z-repeats. Many NEM2 patients have truncating mutations that result in the loss of most of the C-terminus (located within the Z disk), yet few studies have investigated this region of the molecule. Nebulin's C-terminus contains Z-disk repeats that are alternatively spliced (expressed at high levels in muscles with wide Z-disks). We will establish the layout of nebulin in the Z-disk, the location of the Z-repeats, identify protein binding partners, and study the effects of deleting differentially expressed nebulin Z-repeats on the Z-disk structure and function. Capitalizing on our >15-year track record of innovative nebulin research, and utilizing our team of experienced scientists and talented trainees, this proposal sets ambitious goals that are expected to accelerate the understanding of the biology of nebulin, its role in disease, and to identify possible novel therapeutic avenues.

NIH Research Projects · FY 2025 · 2022-08

Project Summary In response to PAR-20-317 (NIA MSTEM: Advancing Diversity in Aging Research through Undergraduate Education [R25]), the University of Arizona Center on Aging (ACOA) proposes to implement the ACOA/ MSTEM THRIVE Program, a substantively focused, structured, longitudinal, mentored research education program targeting University of Arizona (UA) undergraduates (Freshman through Seniors) from groups underrepresented in the health sciences (URHS). Students will be recruited from four pre-health programs in the University of Arizona Health Science Center (UAHS): medicine, nursing, pharmacy and public health. Students will be principally of Latino and American Indian/Native American descent, or from U.S-México border regions. The project is supported by the well documented lack of diversity among the research workforce in aging, the rapid growth of older adults (55 years and older) in diverse groups, and the well documented health disparities in those groups. Consistent with the NIH Strategic Directions for Research--2020-2025 document, research education will focus on the interrelationships of the environmental, social, cultural, behavioral, and biological factors that create and sustain health disparities among older adults. The program will operate within the context of already existing, world-class resources at UA focused on geriatrics/gerontology research and education (ACOA), and existing on-campus programs designed to recruit and promote success among students from groups URHS. The project specific aims are: (1) Develop and sustain undergraduate student interest, involvement, and retention leading to commitment to a research career focused on our theme of Improving health, wellbeing and independence of older adults in diverse groups; 2) Develop, sustain and support undergraduate student engagement in research on our theme by providing students with distinct hands-on experience, requisite knowledge and research skills; 3) Facilitate and sustain undergraduate student progress toward achieving career goals and Cultivate and sustain a cadre of culturally competent research mentors to support MSTEM/ACOA THRIVE students. The innovation of this project lies in our capacity to leverage excellent, already existing on-campus programs designed to recruit and promote success among students from groups URHS with nationally recognized resources and researchers in gerontology/geriatrics, with scientists studying health disparities to improve the health, well-being and independence of older adults in diverse populations. This project is supported within a research-intensive university that is guided by a strategic plan to increase diversity and prepare for an aging world.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Arthrogryposis is present in 1 in 3,000 live births causing joint contractures in both upper and lower limbs. There is no cure making it an unmet medical need. Mutations in the MYBPC1 gene encoding slow skeletal myosin-binding protein C (sMyBP-C), expressed in both slow and fast muscle types are associated with distal arthrogryposis (DA). MYBPC2 encodes for fast skeletal MyBP-C (fMyBP-C) and is found only in fast- twitch muscle. As a myosin-anchored protein of muscle, MyBP-C extends toward actin, positioned centrally in the sarcomere to regulate actomyosin interactions in force development. MyBP-C in skeletal muscle has three major regulators: isoform (slow vs. fast), splice variant (long vs. short sMyBP-C), and posttranslational modification (phosphorylation). sMyBP-C is phosphorylated by protein kinase A (PKA) at its N terminus. The role(s) of sMyBP-C, its phosphorylation and DA mutations in skeletal muscle are not known. Our preliminary studies of sMyBP-C show that binding to actomyosin is dependent on phosphorylation and DA mutations. We have developed innovative biophysical tools that enable evaluation of skeletal MyBP-C structural dynamics, actomyosin interactions in muscle, and effects of phosphorylation and mutations. Our new preliminary studies demonstrate that we have successfully developed fluorescent sensors in N terminal sMyBP-C whose structure and dynamics are sensitive to PKA-mediated phosphorylation and binding to actin. We have also developed inter-molecular fluorescence assays that resolve actin binding between fMyBP-C, long sMyBP-C, and short MyBP-C due to phosphorylation and the presence of tropomyosin on actin. These preliminary results suggest key physiological mechanisms of regulation for the different skeletal MyBP-C and provides additional scientific premise and feasibility for pursuing the proposed studies. Aim 1 will evaluate effects of sMyBP-C binding and DA mutations on interactions with actomyosin, capturing structure and proximities of N terminal sMyBP-C, actin and myosin. Spectroscopic probes will be placed in these proteins and approaches will be employed to detect key conformations in vitro and in situ with wild type and DA mutant sMyBP-C. For fiber experiments, muscle will be isolated from novel sMyBP-C knockout (KO) mice and permeabilized with recombinant sMyBP- C, DA mutants, and muscle protein probes. Samples will be assessed for binding and contractile function. Aim 2 will determine how PKA-mediated phosphorylation of sMyBP-C affects the parameters evaluated in Aim 1. Aim 3 will determine how fMyBP-C affects the parameters evaluated in Aim 1 except using fMyBP-C KO and sMyBP-C/fMyBP-C double-KO mice for fibers experiments. The proposed studies capture structural dynamics and interactions in real time and myofilament space using novel high-resolution approaches. These aims outline a stepwise plan for studying normal and mutant skeletal MyBP-C during the contractile cycle. By monitoring distances between points on proteins and the order (or disorder) of those distances under physiological conditions, mutants can be separated into bins to facilitate targeted mechanistic-based therapies.

NIH Research Projects · FY 2025 · 2022-07

ABSTRACT Many breast cancer survivors (estimated 70% in some studies) experience clinically significant depression and/or anxiety (together psychological distress, hereafter distress) in the months and years after finishing cancer treatments. To address this important need, meditation interventions have been developed to reduce distress for breast cancer survivors, while also improving (or stabilizing) objective indicators of stress physiology (i.e., diurnal cortisol rhythm; DCR). In the last several years, progress has been made to deliver these meditation interventions through digital technology to improve scalability and accessibility (i.e., online and/or through apps) to reduce distress experienced by survivors. Online meditation interventions are especially important because they ensure ongoing viability of a popular integrative modality for many survivors who are unable to engage in face-to-face supportive care activities outside the home. Despite progress, existing online meditation interventions have several limitations that undercut their effectiveness to reduce survivor distress. Online meditation interventions have: 1) centered mostly on mindfulness, which does not actively work to cultivate prosocial and interpersonal relationship capacity and provides limited contemplative skills for survivors who may suffer from feelings of social isolation that contribute to distress; 2) often been asynchronous, with no direct interventionist contact over the course of meditation training; and 3) have often not included survivors' informal caregivers (i.e., adult family members who live with and typically provide half the care for survivors). This research will address these limitations. We will build on the rigor of prior research to reduce survivor distress with a compassion meditation intervention called CBCT (Cognitively-Based Compassion Training) for online synchronous delivery by trained interventionists that is also inclusive of caregivers. CBCT is an 8-week intervention that starts with attention and mindfulness but then builds to contemplation about compassion for the self and others, and is different from other meditation interventions (i.e., mindfulness meditation). The goal of CBCT is to directly enhance feelings of social connection, equanimity, and self-compassion, and reduce negative affect, and through these pathways lessen distress. We seek to determine if CBCT reduces distress and improves DCR for survivors compared to an active attention control (Health Education, HE) when delivered remotely by Zoom to both survivors and caregivers as a dyad (i.e., CBCT for dyads; CBCT-D). We will also test whether or not including caregivers in CBCT-D reduces distress and optimizes DCR more than when survivors receive CBCT by themselves alone (i.e., CBCT for survivors; CBCT-S). If successful, this research will provide evidence regarding efficacy of an online, synchronous dyadic CBCT intervention and the extent to which it does so through the mediational pathway of social connection with caregivers. This research will fill an important gap in evidence for interventions that can reduce distress, an NCI priority for the growing numbers of survivors and caregivers.