Florida State University

NIH Research Projects · FY 2025 · 2021-05

Program Director/Principal Investigator (Last, First, Middle): Ralston, Penny A. The Health for Hearts United Collaborative Project Summary Cardiovascular disease (CVD) is the leading cause of death in the United States, and disproportionately affects African Americans (AAs) who have the highest rates for CVD-related morbidity and mortality in comparison to other ethnic/racial groups. Risk factors for these high CVD rates are related to a variety of factors, including lifestyle. Church-based interventions have been shown to be effective in improving physical health outcomes of AAs. However, a critical barrier to advancing the science of church-based health is understanding the most effective strategies and the extent to which evidence-based health programs can be implemented and maintained by churches themselves The Health for Hearts United Collaborative (HHUC), a community- academic partnership comprised of 45 churches in collaboration with a broader multi-county health coalition, was established after two successful intervention studies to reduce CVD risk in AAs in a two-county area of North Florida, using community-based participatory research approaches. We now seek to use this collaborative environment to investigate implementation of this intervention by the churches themselves as we expand the HHUC. Thus, the proposed project will determine the effectiveness of HHUC implementation strategies in relation to process outcomes and reducing CVD risk in AAs, guided by ecological theory, the Consolidated Framework for Implementation Research (CFIR), and the RE-AIM framework, and using a two- phase approach. The HHUC model currently includes three components: governance structure, annual events, and basic support. Based on observed successes in selected HHUC churches, we propose adding a fourth component that includes one of two possible implementation strategies: 1) an internal champions (IC)-driven strategy that includes two features (leadership development, culturally-tailored planning approaches) or 2) an external change agent (external professionals [EP])-driven strategy without these features. In Phase 1, we will pilot and refine the IC and EP-driven implementation strategies using health leaders from four churches in the two-county area by determining feasibility and acceptability. In Phase 2, we will use an effectiveness- implementation hybrid Type 3 design to evaluate the IC and EP implementation strategies in relation to process outcomes (reach, adoption, implementation and maintenance); and individual health behaviors (food choice, dietary quality, physical activity) and clinical outcomes (BMIs, girth circumferences, systolic and diastolic blood pressure), using congregants ((>18, n=225) in nine churches in the two-county area: three IC treatment, three EP treatment, and three comparison with delayed comparable activities. The findings from this study will inform the expansion of the HHUC and the reduction of CVD risk in AAs, with implications for other communities and regions in the U.S. OMB No. 0925-0001/0002 (Rev. 03/2020 Approved Through 02/28/2023) Page Continuation Format Page

NIH Research Projects · FY 2025 · 2021-05

Project Summary Prediabetes, a comorbidity of obesity and a precursor of type 2 diabetes, affects more than one-half of women over 60 years of age. Obesity has multiple causes; however, it is known that obese insulin resistant individuals have a reduced ability to alter resting and stimulated lipolysis (fat breakdown). This lack of flexibility to respond to stimuli that regulate lipolysis has been attributed, almost entirely through studies in males, to changes in the predominant (catecholamine-mediated) lipolytic pathway. Our published preliminary data demonstrate that acute resistance exercise increases lipolysis in non-obese women. Published data also indicate that resistance exercise, like endurance exercise, increases lipolytic sensitivity in men. However, the alterations in lipolytic response due to resistance, as compared to endurance, training matched for energy expenditure have not been investigated. It is also unknown how training alters lipolysis during general physical activity (walking), which accounts for the majority of activity people engage in during a typical day outside of planned exercise. Furthermore, the lack of prior investigations in this area in women points to the need for resistance training studies of fat metabolism in women to determine if resistance training is as effective as endurance training. Therefore, the overall objective of this study is to compare the effects of 12 weeks of resistance training to endurance training with respect to fat metabolism, with a focus on lipolysis in postmenopausal women with obesity and prediabetes. Our central hypothesis is that both 12 weeks of resistance training and 12 weeks of endurance training will increase lipolytic flexibility. We will compare the effects of endurance training to the effects imparted by calorie-matched endurance exercise training. We will determine with powerful in-vivo microdialysis and stable isotope methodologies the extent to which 12 weeks of resistance training, as compared to calorie-matched endurance training: a) increases physical activity (walking)-stimulated whole-body and regional lipolysis (Aim 1); b) increases local adrenergic regulation of lipolysis in subcutaneous abdominal and gluteal adipose tissue (Aim 2); and c) increases insulin-mediated suppression of whole-body and regional lipolysis (Aim 3) in postmenopausal women with obesity and prediabetes. Secondarily, fat oxidation, lipogenesis and adipogenesis in adipose tissue, as well as lipolytic activity in skeletal muscle, will also be studied to develop a global understanding of fat metabolism response to resistance exercise training. In addition, we will investigate the influence of resistance and endurance training on glucose profile under laboratory as well as free-living conditions, as poor glucose control is linked to the aberrant lipid metabolism commonly associated with obesity. These studies will provide a greater understanding of how these exercise modalities affect metabolism in women with obesity and prediabetes, allowing practitioners to make more evidence- based exercise prescriptions intended to improve body composition, glycemic control, and weight management.

NIH Research Projects · FY 2025 · 2021-02

PROJECT SUMMARY/ABSTRACT Alzheimer’s disease is devastating for individuals and society. Impaired learning and memory, particularly in the context of spatial navigation, is one of its early and major symptoms. Similarly, rodents recapitulating aspects of Alzheimer’s disease also exhibit early impairments in spatial navigation. A preponderance of evidence suggests abnormal cortical-hippocampal communication in humans with Alzheimer’s disease. Hippocampal-cortical interactions during sleep are thought to be critical for consolidation of newly acquired memories. However, no studies have assessed these brain dynamics during sleep in rodents modeling Tau and amyloid beta (Aβ) aggregation aspects of Alzheimer’s disease. Thus, the proposed research will explore the functionality of brain dynamics during sleep in the hippocampal-PC network in animal models of Tau and Aβ aggregation (TAβA). To do this, we will use a triple transgenic mouse where three major genes associated with familial Alzheimer’s disease are expressed leading to TAβA. This mouse model mimics plaque and tangle pathological hallmarks of the disease, with a distribution pattern similar to human patients, including synaptic changes in the limbic system. In addition, all findings will be confirmed in a transgenic rat with Aβ accumulation, plaque formation, tau accumulation, cell loss, and spatial memory impairments. Specifically, we will: 1) assess the relationship between spatial learning and memory, as well as brain dynamics during sleep, both within and across the hippocampus and cortex; 2) use a novel targeted optogenetic approach to functionally dissect the relative contributions of TAβA in the hippocampus to impaired hippocampal-cortical coupling during sleep and impaired spatial learning. 3) test the efficacy of a non-invasive visual stimulation approach, known for clearing cortical TAβA, to relieve impaired hippocampal-cortical coupling during sleep and impaired spatial learning. This project will provide insight into the normal function of a circuit that is dysfunctional in Alzheimer’s disease and allow us to probe dysfunction in this circuit that emerges in very early stages of disease progression in rodents modeling TAβA aspects of Alzheimer’s disease. This research will allow us to begin understanding changes in this network which may underlie the emergence of cognitive impairments observed in Alzheimer’s disease and begin testing the efficacy of a non-invasive treatment for reversing the functional brain abnormalities and impaired cognition.

NIH Research Projects · FY 2026 · 2021-01

Abstract Functional reconstitution of membrane proteins has been the major roadblock for the application of NMR and other biophysical techniques to investigate their high-resolution dynamic structures in a native membrane environment. In this application, we propose to develop approaches to enable high-resolution structural studies of membrane proteins and protein-protein complexes by a variety of biophysical techniques. We will develop nanodisc technology for detergent-free direct extraction and functional reconstitution of membrane proteins for structural studies of a variety of membrane proteins including single-pass transmembrane proteins (such as mammalian cytochromes and heme oxygenase) and integral membrane proteins (including GPCRs and Guanidine exporter). Synthetic polymers developed in our laboratory exhibit the ability to form nanodiscs with easily controllable sizes (from ~8 to ~60 nm diameter), are stable against pH and divalent metal ions and capable of directly extracting membrane proteins. Our preliminary results demonstrate that these nanodiscs (<20 nm diameter) and macro-nanodiscs (>20 nm diameter) represent an exciting system for solution and solid-state NMR studies of membrane proteins. We also propose to use the newly developed nanodisc technology and NMR approaches to investigate the structural interactions of mammalian cytochrome-P450 (P450) with its redox partners (P450-reductase (CPR) and cytochrome-b5 (b5)) to better understand how redox partners regulate P450 catalysis and how P450s metabolize chemically diverse substrates. The structural aspects pertaining to the catalytic activity of P450s continue to remain elusive due to a lack of high-resolution structures in their full-length, active forms. Presently, structural studies of P450s are restricted to various truncated mammalian and water-soluble bacterial P450 homologs. In this study, we will investigate the structure, dynamics and transmembrane domain orientation of full-length mammalian P450s (2B4, 3A4 and 3A5 isoforms) alone and in complex with its redox partner b5 and CPR, incorporated in nanodiscs, using a combination of high-resolution solution and solid-state NMR techniques. We will also investigate the ternary P450-b5-CPR complex in nanodiscs in the presence of substrates to elucidate the molecular origin of the strikingly different effects CPR and b5 have on P450 2B4 catalysis. The outcome of the proposed studies on P450-redox complexes will provide structure and dynamics/function principles regulating P450 metabolism of a wide variety of substrates. The results obtained from this study will also be useful to design potent drugs to ultimately treat and prevent diseases including cancer.

NIH Research Projects · FY 2025 · 2021-01

Project Summary The project addresses a common challenge in the remediation of groundwater contaminated with chlorinated volatile organic compounds (CVOCs) and 1,4-dioxane. CVOCs include chlorinated solvents, such as trichloroethylene (TCE) and 1,1,1-trichloroethane (1,1,1-TCA), and their degradation products. Many CVOCs and 1,4-dioxane are known or potential human carcinogens and on the Substance Priority List (SPL) for Superfund sites. CVOCs bioremediation under anaerobic conditions (i.e. reductive dechlorination) is well established. However, bioremediation of mixtures of CVOCs and 1,4-dioxane is not yet feasible due to at least the following three obstacles: 1) low biodegradability of 1,4-dioxane at environmentally relevant concentrations, 2) requirement for aerobic conditions for 1,4-dioxane metabolism but anaerobic conditions for most CVOCs metabolism, and 3) inhibition of 1,4-dioxane biodegradation by CVOCs. This project proposes the following combined remediation approach to address these challenges: first, an innovative macrocyclic material approach to selectively adsorb CVOCs and promote the growth of dechlorinating biofilm on the material surface to anaerobically biodegrade CVOCs. After the CVOCs treatment, another type of innovative macrocyclic material as an effective and selective sorbent for 1,4-dioxane sustains biofilms consisting of a highly efficient culture to aerobically metabolize 1,4-dioxane. The macrocyclic molecules, which comprise repeating cyclic oligomers with unique geometry and internal chemistry, form specific host-guest complexes with only selected guest molecules (i.e., 1,4-dioxane or CVOCs). A highly efficient 1,4-dioxane-metabolizing culture (previously established) is much more effective at low, environmentally relevant concentrations compared to all others reported in literature. To understand the mechanisms of how the novel sorbents enhance bioremediation and to demonstrate the feasibility of the proposed remediation approach, the researchers will conduct the following work: 1) Computational study, synthesis, and characterization of novel macrocyclic materials. Two sorbents, one that selectively and reversibly adsorbs CVOCs and another that selectively adsorbs 1,4-dioxane will be optimized for use in the bioremediation studies. 2) Mechanistic study of the highly efficient 1,4-dioxane-metabolizing culture. Key microorganisms responsible for the high affinity to 1,4-dioxane in the mixed culture will be isolated and investigated for their degradation intermediates, pathways, and kinetics. 3) Elucidation of interactions among contaminants, microbial cultures, and the novel sorbents. To achieve this, completely mixed flow experiments will be performed, and they will be coupled with mathematical modeling that incorporates phenomena of both sorption and biodegradation in biofilms. 4) Proof-of-concept column studies for bioremediation of CVOCs and 1,4-dioxane mixtures. Two long-term column studies will be performed: ex situ treatment of 1,4-dioxane and in situ bioremediation of CVOCs and 1,4-dioxane mixture in series. Performance objectives will be Maximum Contaminant Levels for CVOCs and the Health Advisory Level for 1,4-dioxane (0.35 µg/L).

NIH Research Projects · FY 2025 · 2021-01

Project Summary The long term goal of this research project has been and continues to be an understanding of the molecular mechanism of muscle function. The existing project, funded continuously since 1983, has evolved in parallel with the capabilities of both microscopes and methods for 3-D image reconstruction from electron microscopes. Originally focused on understanding myosin-actin interactions in situ in muscle using chemically fixed, plastic embedded and sectioned muscle, it now proposes to take advantage of the resolution revolution in cryoEM to study the major structural elements of the muscle at the highest resolution possible using isolated components, followed by a return to imaging actin-myosin interactions in situ in frozen live muscle cells. Striated muscles have four major components: actin-containing thin filaments, myosin- containing thick filaments, a Z-disk to crosslink antiparallel thin filaments and a connecting filament to link the thick filaments to the Z-disk. The least understood of these four elements are the thick filament, the Z-disk and the connecting filament, whose interactions with the thick filament and Z-disk are its least understood elements. Thus, our study of Z-disk and thick filament can make major contributions to an understanding of all three elements. Following a major breakthrough of ours that showed that coiled-coil tail domain of myosin can be imaged at subnanometer resolution, even near atomic resolution, the project concentrates initially on subnanometer resolution imaging of thick filaments from several species, to examine the generality and structural conservation of the “curved molecular crystalline layers” across species and muscle types. The project will utilize the fruit fly, Drosophila melanogaster, to investigate how genetic removal of certain component proteins affects how the myosin tails interact with non-myosin proteins to affect thick filament properties. We will utilize mutations in the myosin tail of Drosophila that correspond to established disease causing mutations in human striated muscle. The Z-disk will be studied using methodology developed in our lab to isolate Z-disks from invertebrates applied to determination of the Drosophila melanogaster Z-disk. The experimental system will facilitate decoration of the Z-disks with various signaling proteins. Ultimately, the utility of these studies on components needs development within the myofibril. We will investigate by cryoelectron tomography frozen-hydrated myofibrils of Lethocerus and Drosophila thinned using FIB/SEM in states produced using various nucleotides to see how myosin heads interact with the thin filament in situ. Ultimately, we will apply what we have learned methodologically from these myofibril studies to studies of live cultured smooth and cardiac muscle cells fast frozen, thinned via FIB/SEM to visualize active interactions between thick and thin filaments. This work will open to future structural investigation all the structures present in a muscle cell within their natural context.

NIH Research Projects · FY 2024 · 2021-01

Project Summary Tyrosine O-sulfation, i.e., transfer of a sulfonate group to tyrosine amino acid residues in proteins, is a widespread posttranslational modification (PTM) in eukaryotic cells with a variety of known functions in health and disease, including receptor binding, viral replication, inflammation, and retinal function. The enzymes that catalyze tyrosine sulfation are located in the Golgi apparatus. The function of this organelle is to ensure that correct protein modification occurs and to package proteins into vesicles for export to the cell surface, or the extracellular environment. Because proteins must typically enter the Golgi to become sulfated, most known sulfoproteins are secreted proteins or membrane proteins. Mass spectrometry (MS) is a powerful tool for global PTM analysis in cells and tissues; however, large scale analysis of tyrosine O-sulfation has not been feasible, due in part to its labile nature in the gas-phase environment of a mass spectrometer, and in part due to the lack of appropriate data analysis strategies. In MS experiments, proteins are typically digested into smaller peptides, which are ionized, detected, and fragmented to deduce sequence information. When measuring protein phosphorylation, another rather labile PTM known to regulate Golgi disassembly and reassembly during cell division, in interphase vs. mitotic Golgi, we found that tyrosine O-sulfation was co-enriched. This discovery is not surprising because the chemical properties of sulfation (O-SO3) are similar to phosphorylation (O-PO3H). However; high mass accuracy measurements are required to deduce the small mass difference of 0.0095 Da between these two PTMs. Even as such high performance measurements are becoming more routine, standard database search tools typically do not identify protein sulfation because this PTM is completely lost during analysis. We found that open database searching was able to overcome this problem and, thus, we were able to accomplish identification of a number of novel sulfoproteins in rat liver Golgi. While an exciting advance, the exact location of O-sulfation within proteolytic peptides could not be directly measured. In Aim 1 of this proposal, we seek to develop improved methods for detection of intact sulfopeptides by MS, including elimination of competing phosphorylation, determination of peptide sequence effects, implementation of stabilizing adducts, and conditions that selectively dissociate sulfopeptides. To further allow sulfate site localization, in Aim 2, we seek to develop technologies for fragmenting sulfopeptides while retaining sulfate in fragment ions. These approaches include negative ion mode free radical initiated peptide sequencing, which allows sulfopeptides to enter the mass spectrometer as more stable anions, and the development of “smart” data acquisition strategies for improved electron transfer dissociation. The final Aim 3 seeks to apply these improved approaches for comprehensive analysis of the Golgi sulfoproteome in cells and animal tissue, particularly under perturbed Golgi conditions, which are expected to alter sulfation. These types of measurements will provide transformative information regarding the regulatory roles of tyrosine sulfation and its impact on cellular function.

NIH Research Projects · FY 2024 · 2020-09

Summary/Abstract Cognitive, emotional, interpersonal, and physical functioning are profoundly impacted by Alzheimer's disease (AD) and related dementias. AD poses enormous public health and societal challenges, and the number of affected individuals is expected to rise if an effective intervention to stop or at least slow AD progression is not found. Research on the biological and psychosocial mechanisms that lead to dementia is essential to identify appropriate targets for interventions. The scientific premise of the proposed study is based on robust evidence for an association between long-standing personality traits and dementia-related outcomes. The mechanisms through which these traits lead to cognitive changes, however, are not well understood. Furthermore, with the onset and progression of AD, personality turns from a risk factor to a clinical sign of the disease, but the timing and trajectory of such changes are not well understood. By integrating multiple theoretical perspectives (five- factor model, lifespan models of personality and health, pathoplastic approaches to psychopathology, and cognitive reserve), the proposed research will test three innovative aims. The first aim is to advance a mechanistic understanding of the association between personality and dementia. We will test neurobiological (e.g., MRI brain region volumes and white matter integrity), physiological (e.g., cortisol and cardiovascular burden), behavioral (e.g., physical inactivity and smoking), and psychosocial (e.g., depressive symptoms and loneliness) factors in the pathway from personality to cognitive decline and dementia. The second aim is to test whether personality uncouples AD neuropathology from clinical dementia: We expect that, in the presence of neuropathological changes, a resilient personality profile (low neuroticism, high conscientiousness) will delay the time to onset of mild cognitive impairment (MCI) and dementia. The third aim is to identify the timing and trajectory of personality change during the prodromal phase of AD and related dementias. To address the study aims, we will leverage data from the Baltimore Longitudinal Study of Aging (BLSA). This ongoing prospective study include measures of the five major personality traits, along with in-depth assessments of relevant risk factors and AD biomarkers, and they involve long-term follow-ups (up to 40 years of serial assessments of personality in the BLSA). By leveraging rich prospective data, the proposed project will identify the biological and psychosocial mechanisms that underlie personality-based vulnerability and resilience to dementia and identify inflection points for personality change in the earliest symptomatic phase of the disease.

NIH Research Projects · FY 2020 · 2020-04

Amyloid diseases are a group of diseases in which more than 30 proteins are known to aggregate and cause degenerative diseases. These diseases have a common underlying amyloid fibril accumulation in specific organs such as the brain, pancreas, and heart. Neurodegenerative diseases such as Alzheimer’s disease (AD) and Parkinson’s disease (PD) are the examples of highly progressively debilitating age-related brain diseases under this category. Type 2 diabetes mellitus and cardiac amyloidosis belong to the extracerebral amyloid diseases that have the worst prognosis without treatment. These diseases have sporadic and familial origins, infectious forms such as the spongiform encephalopathies, and localized as well as systemic forms such as the transthyretin amyloidosis. Despite making a significant progress to understand the pathogenesis of these diseases over the last 3 decades, most clinical trials have failed to produce successful therapies so far for the neurodegenerative diseases (e.g., AD and PD). Furthermore, diagnoses of these diseases at the initial stages are difficult and no definitive laboratory tests exist for most of these neurodegenerative diseases. The diagnostic difficulties and clinical trial failures clearly indicate poor or incomplete understanding of the origin and pathomechanisms of these diseases. Also, it is possible that these diseases may have different origins or causes and disease progression pathways. Perhaps these diseases cross a much bigger basic science, engineering and clinical science disciplines, and thus a much broader inter-disciplinary team of research scientists should come together to make a better understanding of these diseases. This conference is being organized to educate junior faculty members and graduate students from diverse disciplines what was understood thus far about these diseases on diverse topics such as misfolding and aggregation of amyloid proteins, self-assembly process, amyloid toxicity, cellular studies to identify molecular mechanisms, diagnosis, biomarker and inhibitor developments, and treatment options. The organizers have identified speakers in multiple disciplines who have lately come up with novel and cutting-edge approaches to better understand or diagnose these diseases. With the above background, the primary goal is to enable junior faculty members, graduate students and post- doctoral fellows to attend this conference so that they will be better informed of these diseases that will motivate them to target their research focus on the diseases. The secondary goals include deliberations on: 1) as to how to bridge the gap between in vitro and in vivo studies and enhance collaborations among researchers working in these areas, 2) whether basic research be better focused to identify and characterize the most toxic early aggregates under in vitro and cellular conditions, 3) whether the identified amyloid species be tested in animal models for the development of biomarkers, 4) development of new approaches for in vitro and in vivo animal models, and 5) as to how to bridge the “translational” gap for improved identification of newer therapeutic targets for the neurodegenerative diseases.

NIH Research Projects · FY 2025 · 2020-01

Cognitive control refers to the ability to guide behavior in an intentional and goal-directed manner amidst competing demands. This hallmark of human cognition is supported by the prefrontal (PFC) and posterior parietal cortices (PPC), areas that have undergone extensive evolutionary expansion. The PFC and PPC are central to integrating the present context with plans for the future in order to guide intentional behavior. Accordingly, dysfunction of these regions leads to a wide-variety of deficits including inflexibility, inattention, impulsivity, and disorganization. Such cognitive deficits are evident in numerous psychiatric and neurological disorders such as schizophrenia, attention-deficit hyperactivity disorder, substance addiction, mood disorders, Parkinson’s disease, Huntington’s disease, stroke, and traumatic brain injury. However, impairments in cognitive control are particularly challenging to treat in part due to insufficient mechanistic understanding of the PFC, PPC, and their interactions. To understand and treat disorders of higher-level cognition we need to detail the directed interactions of the PFC and PPC, elucidating functional chains among brain regions and behavior. However, elucidating directed interactions in humans is challenging given limitations of available techniques. Animal models may not translate to the PFC and PPC-mediated abilities that are exceptional in humans. As a result, how the directed interactions of the PFC/PPC mediate higher-level cognition and how they can be manipulated to model dysfunction and move towards rehabilitation remains unclear. This proposal aims to fill this gap using a combination of techniques. Functional magnetic resonance imaging (fMRI) will be coupled with computational techniques to model how directed PFC/PPC interactions support cognitive control. Chief among the interests of this proposal are identification of putative hierarchical organizations with the PFC and PPC that are symbolized by asymmetries of directed influence. Areas at the apex of such hierarchies are predicted to exert widespread influence over other brain areas and cognition. Such apical areas would therefore serve as important biomarkers to monitor for disorder progression, and targets for treatment. Modeled apical roles will be causally validated using interleaved transcranial magnetic stimulation (TMS) and fMRI by examining the impact of focal stimulation on downstream brain areas and behavior. Both continuous theta-burst TMS (cTBS) and intermittent theta-burst TMS (iTBS) will be employed with putative inhibitory and excitatory effects, respectively. It is predicted that cTBS will impair behavior, serving as a model of dysfunction, while iTBS will enhance behavior, serving as a road towards treatment. Aim 1 will estimate hierarchical models in the PFC, validate these models using cTBS, and start the path towards treatment using iTBS. Aim 2 will apply a similar logic to the PPC and contrast the relative efficacy of PFC vs. PPC TMS. Collectively, these aims will provide directed models of PFC/PPC interactions supporting cognitive control, and causal data regarding how targeted manipulation of these networks can hinder or improve control.

NIH Research Projects · FY 2026 · 2017-09

PROJECT ABSTRACT Natural products continue to have a disproportionate impact on how we understand and treat disease. These biologically pre-validated therapeutic leads have inspired ~50% of FDA-approved drugs, a fact underscores the importance of studying biogenic chemotypes with unique activity in cell culture. Inspired by the rich opportunities for natural products at the interface of chemistry, biology, and medicine, this proposal focuses on a family of terpenoids distinguished by a fused [5-8-5] carbocyclic molecular framework. The flagship members of this family possess impactful, albeit divergent, pharmacological profiles. For example, fusicoccin A stabilizes 14-3-3 protein- protein interactions (PPIs), ophiobolin A is a cytotoxic membrane disruptor, and bipolarolide A inhibits HMG-CoA reductase. The specificity of these terpenoids for their diverse biological receptors is determined by the identity and arrangement of substituents surrounding a common 5-8-5 substructure. Given the complex conformational dynamics of cyclooctanoids, we postulate that the groups flanking the central eight-membered ring modulate the overall molecular shape. This potentially programmable feature might allow this terpenoid family to interface with diverse biological receptors. Thus, the [5-8-5] ring system appears to be a privileged scaffold for the design of novel therapeutics. However, existing approaches to this family of terpenoids rely on convergent, target-specific chemistry. Viewed through the lens of drug discovery, the lack of chemistry to diversify the [5-8-5] motif is the main impediment to understanding and advancing the pharmacology of this natural product family. During the past grant period, we developed a synthetic platform to prepare and edit the fusicoccin scaffold. We now seek to extend this chemistry in several directions. In Aim 1, we will exploit our platform to establish the features of fusicoccin A that impart selectivity for individual 14-3-3 PPIs. In doing so, we bring a unique chemical perspective to the long-standing problem of selectively modulating the 14-3-3 interactome. In Aim 2, we will extend our synthetic strategy to capture ophiobolin A and bipolarolide A. These isomeric sesterterpenes feature peripheral and skeletal modifications to the 5-8-5 scaffold that cannot be addressed by existing chemistry. We outline an enabling synthetic entry point to these molecules from a common 5-8-5 intermediate. Thus, successful completion of the proposed research will establish a divergent platform to chemically redesign [5-8-5] terpenoid chemotypes. It will also establish whether the fusicoccins can be exploited to interrogate specific components of the 14-3-3 interactome in cell culture. This research will signifyingly broaden access to [5-8-5] terpenoid natural products with diverse pharmacological profiles and shed light on how to chemically evolve this chemotype toward a specific functional endpoint.

NIH Research Projects · FY 2025 · 2017-06

PROJECT SUMMARY Dietary indices and scores provide a comprehensive and robust approach for nutritional exposure assessment, especially in relation to chronic disease risk. While dietary supplement (DS) use is very common among the U.S. population, with over 52% of adults and 33% of children routinely taking one or more (predominantly nutrient containing) products, no existing metrics assessed intakes from DS in addition to those from the diet. For this reason, we developed the Total Nutrient Index (TNI), a measure designed to evaluate total usual micronutrient intakes, inclusive of intakes from foods and DS, relative to the Dietary Reference Intakes for eight micronutrients identified as under consumed among the U.S. population by the 2015-2020 Dietary Guidelines for Americans (DGA) in the first period of this award. We examined the relative validity of the TNI compared to biomarkers of nutritional status among a nationally representative sample of U.S. adults and determined that 1) the TNI is a useful metric for total micronutrient exposure assessment, and 2) that the TNI yielded higher correlations with nutritional biomarkers when compared with those obtained from dietary intake alone. However, with the emergence of the precision nutrition era, understanding how unique combinations of nutrients and specific amounts of these nutrients can optimize the nutritional and health status of different population subgroups is of utmost importance. Accordingly, in the present competing continuation proposal, we intend to update the TNI to accommodate a life stage approach, to utilize machine learning techniques to identify critical nutrients and in what combinations optimally benefit the health and nutrition of individuals across the life course, and relate the TNI to risk factors, health outcomes, and mortality. In Aim 1, we plan to develop and validate the TNI-2, an updated extension of the TNI that is tailored to different life stages and includes nutrients and food components of

NIH Research Projects · FY 2026 · 2017-04

Project Summary Abstract The 26S proteasome conducts most regulated protein degradation and eliminates toxic proteins from cells. The proteasome is a validated anti-cancer target, and holds substantial promise as a target for treatment of neurodegenerative disorders and some infectious diseases. Our long-term goal is to understand how the three major complexes of the proteasome—the lid, base, and core particle—engage and communicate within and between one another. We have thus far made significant progress toward this goal and have developed a number of novel tools and reagents that have furthered our understanding of intra- and inter-complex communication. Conceptual advances have included: i) discovery of additional conformational states of the yeast proteasome relevant to substrate catalysis; ii) demonstration that six highly similar ATP-hydrolyzing subunits differentially influence the activation state of the proteasome; iii) discovery of a link between the conformational state of the proteasome and release of a dedicated proteasome assembly chaperone; and iv) and the finding that proteasomal subcomplexes disengage one another prior to their destruction by autophagy. A paradigm emerging from this initial budget period is that rather small binding events or molecular movements are transmitted, often over long distances, to enact largescale conformational changes. Understanding how such local events are amplified and transmitted to distant areas of the proteasome to coordinate assembly and catalysis is thus a critical knowledge gap. In this first renewal, we propose three Aims that explore examples of this paradigm newly discovered by us during the initial budget period. Together, they will push our knowledge of proteasome dynamics and inter-complex communication into new arenas. In the first, we will use newly developed FRET-based kinetic assays to decipher how local changes to the lid-base interface regulate the timely binding and release of dedicated assembly chaperones from nascent proteasomes. In the second, we will explore a surprising allosteric conduit originating from the substrate unfolding center of the proteasome that regulates the stability between two key subcomplexes. In the third Aim, we will investigate an unusual eukaryotic proteasome from a poorly studied human parasite from the phylum of Microsporidia. Microsporidia lack several proteasome subunits that normally span a key inter-complex interface. The missing subunits contain several small sequence elements with essential roles in assembly and catalysis in other eukaryotes, so exploring these unusual proteasomes will thus reveal both conserved and unique elements of inter-complex communication. These studies are anticipated to produce important insights into the engagement and communication between the proteasomal subcomplexes, significantly advancing several aspects of proteasome biology and drug discovery. Further, microsporidia are NIH priority pathogens of interest for which no broadly effective treatments exist. Insights into microsporidial proteasomes thus may reveal new targets to combat certain parasitic infections.

NIH Research Projects · FY 2026 · 2017-04

Project Summary Alzheimer’s disease is prevalent at the end of life and remains the only leading cause of death without a cure or way to stop or significantly slow its progression. Prevention remains the best hope for reducing risk of Alzheimer’s disease in older adulthood. Given that Alzheimer’s disease has a complex etiology, with risk factors that range from genetics to the environment, multipronged approaches to prevention will likely be needed for an intervention to be broadly effective. Among the psychosocial risk factors for Alzheimer’s disease, personality traits have emerged as consistent predictors of cognitive health across adulthood. Specifically, higher neuroticism (the tendency to experience negative emotions and vulnerability to stress) and lower conscientiousness (the tendency to be organized, disciplined, and responsible) are associated with worse performance on cognitive tasks, more subjective cognitive complaints, and greater risk of Alzheimer’s disease and related dementias. Even after diagnosis, these traits are associated with behavioral and psychological symptoms at the end of life. Lifespan models of personality and health indicate that personality contributes to long-term health outcomes through both behavioral and clinical pathways. Missing from these models, however, are the antecedents of personality, novel mechanisms that go beyond behavioral and clinical risk factors, and how informant ratings of personality and cognition provide unique information about the target’s cognitive health. This work builds on the success of our previous award that found that personality is shaped by socioeconomic factors and that personality is one mechanism in the pathway from childhood socioeconomic status to adult cognitive health. The purpose of this project is to expand consideration of advantages and disadvantages experienced across the lifespan to include other domains to better understand how the accumulation and interplay of such factors across childhood and adulthood shape adult personality traits. This project will further evaluate socioemotional health and behavioral life skills as novel pathways from personality to cognitive health, which are hypothesized to be mechanisms that go beyond traditional behavioral and clinical risk factors. Finally, this project will also include informant ratings of personality and cognition as an additional source of information that provides unique information about the target’s health. We will address these aims in an established, ongoing longitudinal cohort study. The ultimate goal of this work is to develop a personality-informed intervention to support healthier cognitive aging and reduce risk of Alzheimer’s disease. We seek to build a robust and replicable evidence base for a lifespan model of personality and cognitive health that includes antecedents of personality and mechanisms in this pathway, as a step toward this goal.

NIH Research Projects · FY 2025 · 2017-01

This LD Hub P20 renewal entitled, “Determinants of phenotypes within the word reading (dis)ability population: The impact of varied language experiences and child attributes on emerging reading skills” responds to the NICHD invitation for LD Innovation Hubs, FOA’s (RFA-HD-22-005). The overarching goal of this LD Hub is to continue to lay the foundation for a generation of research that situates educational practices in a novel theory of individual word reading development. The knowledge and product generated from this Hub will be used to inform future behavioral, computational, and neurobiological studies examining the development of word reading skills and will be used to align theories describing the relations between child- and word-attributes that explain individual differences in word reading more closely with the educational challenges confronting educators of typically developing (TD) and more specifically children with reading disability (RD). The proposal addresses the second priority of the RFA; namely “pushing innovation”, by exploring new and complex behavioral phenotypes of RD that vary as a function of child experience and cognitive ability across linguistically varied samples of learners. Our LD Hub adopts an interdisciplinary approach to developing the foundational and translational research needed to better understand the general development of item-level word reading skill in a large portion of the English language, explore important differences in word reading development across TD and RD populations, and examine the interactions between child- and word-attributes that explain individual differences in word reading development. The overall specific aims for the Hub include: (1) expand our understanding of the basic mechanisms undergirding word reading development in English; (2) increase the scientific infrastructure for research on reading disability by establishing a publicly available database (developmental English Lexicon Project) incorporating all of the data collected in the research project; (3) maintain and expand the Hub’s multidisciplinary team of experienced and early researchers with expertise spanning educational, computational, and neurobiological research; and (4) strengthen the scientific workforce by providing career enhancing opportunities for early career scientist. In achieving these aims, we will create a research team, a body of empirical knowledge, and a theoretical framework setting the stage for (i) better educational practices, particularly related to RD; (ii) translational research on related topics such as literacy acquisition by TD and RD populations; and (iii) a new generation of theories embracing individual differences and strongly tied to the neurobiological bases of learning.

NIH Research Projects · FY 2025 · 2012-07

The Integrated Clinical Neuroscience (ICN) Training Program, operating within the Clinical Psychology and interdisciplinary Neuroscience programs at Florida State University for the past 9 years, seeks to train the next generation of investigators to become leaders in translational research and make major advances in several areas of psychopathology characterized by dysregulated behaviors including eating disorders, depression, anxiety disorders, suicidality, post-traumatic stress disorder, and substance use disorders. These problems are associated with significant psychological and medical morbidity, elevated mortality, and high economic burden. This underscores the need for research that translates the neural mechanisms underlying normal and abnormal behavior in animals into clinical studies of the causes and treatment of mental disorders. However, segregation of doctoral training in Clinical Psychology and Neuroscience, with separate courses, lab experiences, and exposure to outside speakers, impedes new scientists’ preparation to undertake translational approaches in their own research. The ICN Training Program was designed to break down these barriers. Here, we seek continued funding to provide integrated instruction, research experience, and mentorship to 4 predoctoral ICN trainees earning PhDs in Clinical Psychology and Neuroscience through 1) cross-area courses, 2) cross-area lab rotations, 3) a Special Speaker Series in which trainees present their research to invited scholars conducting translational science, and these scholars present their work to trainees, our participating Departments, and the community, 4) presentation of research at conferences and in published papers, and 5) instruction in grant writing, data science, and cross-area instruction in the responsible conduct of research. Predoctoral trainees apply for the ICN Training Program by describing research they plan to conduct in collaboration with their primary advisor and cross-area mentor, classes they will complete, and how this cross-area exposure will contribute to their career development as translational scientists. Students are typically appointed for two years beginning in their 3rd to 4th year to ensure selection of the most promising trainees who have completed basic program requirements and established research interests that can be extended by a cross-area lab rotation. Our training model has supported a total of 19 trainees. Among the 13 ICN trainees who have completed the Ph.D., 92% have continued in research careers (with 4 currently in tenure-track positions) and 77% have received highly competitive fellowships and independent research grants. Training faculty, selected for their cross-area connections in research addressing dysregulated behaviors, continue to excel in securing grant funding and training students for research careers with strong publication records. Continuation of the ICN Training Grant for an additional 5 years will build upon our accomplishments and ensure that a third cohort of Ph.D. students is ideally positioned to initiate cutting-edge translational research to address mental disorders characterized by dysregulated behaviors.

NIH Research Projects · FY 2025 · 2008-04

The mechanisms that synchronize hormone secretion across millions of islets of Langerhans in the pancreas are unknown. The long-term goal of the Roper laboratory is to decode cellular communication to enable understanding of normal biological function and disease progression. The objective of this proposal is to identify the mechanisms that generate synchronized rapid and ultradian insulin and glucagon oscillations from multiple islets. The central hypothesis is that multiple mechanisms working in concert produce both rapid and ultradian oscillations of hormone release. The rationale for performing this work is that a thorough understanding of the dynamics of glucose-regulatory hormone secretion will lead to the design of therapeutic approaches that alleviate the complications associated with diabetes and other metabolic diseases. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Determine the effect of time delays and dual entrainment on insulin synchronization, and 2) identify glucagon secretion and synchronization dynamics. Under the first aim, two methods for inducing islet synchronization will be used together, one a negative insulin/glucose feedback loop with time delays, and the other, pulsatile activation of M3 receptors. To accomplish this aim, a high-speed method for insulin measurement will be developed using droplet microfluidics and will be used for testing a range of time delays and the ability to perfuse multiple secretagogues in parallel. In the second aim, glucagon secretion will be measured for the first time from single islets of Langerhans using a homogeneous time resolved fluorescence assay using a microfluidic system. With this method, we anticipate observing single islet glucagon secretion dynamics and will parallelize the method to discern how glucagon pulses are synchronized across multiple islets. The proposed research is innovative because the microfluidic systems and measurement approaches developed in this proposal will allow rapid and ultradian oscillations of islet secretion to be observed for the first time. These results will provide a significant increase in the knowledge of islet regulation, which is crucial for fully understanding the mechanism of glucose homeostasis and how it goes awry in metabolic diseases. Ultimately, this knowledge has the potential to guide therapeutic development for reducing the problems associated with unregulated glucose levels in type II diabetes.

NIH Research Projects · FY 2025 · 2006-07

The long-range objective of the Learning Disabilities Translational Science Collective is to build on successes of the Florida Learning Disabilities Research Center to explicitly address the research to practice gap associated with the identification and treatment of children with specific learning disabilities (SLD) in word reading and reading comprehension. SLDs are a significant public health concern, with many children experiencing difficulty with reading, writing, and language skills in school. It is estimated that as many as 15-20 percent of the population experience effects of SLD. Without effective identification and treatment, individuals with SLD are more likely to drop out of school, have difficulty with employment, and encounter other challenges in daily life. To address this health disparity, the Center is organized around the translational science continuum and the concept of multi-disciplinary team science as a necessary approach to tackle this complex public health problem. This Center has four research projects and three cores (administrative, data management, and engagement) that work synergistically to address applied research questions about SLD. The research projects represent the translational science continuum (T1-T4). Project 1 addresses identification, classification, and risk prediction of SLD. Project 2 (high risk) investigates the genetic and environmental influence on the responsiveness to intervention for children who show early signs of reading risk. Project 3 (engagement) uses an implementation science approach to better understand how to develop sustainable process for assessment systems in authentic school settings. Project 4 will utilize population level screening data to better understand classification systems at scale. Every project includes engagement with early career researchers and community members to advance research and to enhance the translation of findings into practice to benefit children.

NIH Research Projects · FY 2025 · 1995-07

Summary This Chemosensory Training Program (CTP), operating within the interdisciplinary Program in Neuroscience at Florida State University, is a continuing application in its 30th year. The CTP Program is geared to train the next generation of researchers to become leaders in basic neural mechanisms of chemosensory systems interfaced with behavior. The program prepares 4 pre- and 1 post-doctoral trainee for research careers focused on olfactory and gustatory senses in context. One important context is the regulation of food intake and metabolic state, dysregulation of which can lead to obesity and diabetes, or anorexia. The powerful links between chemosensory systems and brain circuitry associated with emotional, motivational, and neuromodulatory processes requires a wide perspective for full understanding. The broad long- term objective is to provide the basic neuroscience platform upon which clinical understanding of chemosensory disease is built using a wide spectrum of experimental approaches including molecular neurobiology, neurophysiology, biophysics, psychophysics, and behavioral analysis. The strength of the CTP program that anticipates to provide 2 to 3 years of training for approximately five post- and fifteen pre-doctoral scholars, respectively, is the close guidance of trainees by expert faculty whom are accustomed to productive collaborations fostered from a wealth of historical chemosensory knowledge that shapes cutting-edge investigations for training. Trainees have access to state-of-the-art custom-designed chemosensory equipment, technical support staff, and modern building infrastructure to perform their research. Value- added activities include – 1) chemosensory tutorials (hands-on lab practicum, rigor and reproducibility training), 2) “scholar in practice” (career internship with former alumni), 3) chemosensory retreat (research progress/sharing and mentor/mentee career development), 4) structured oral, written, and analysis skill building and feedback, 5) depth of a continually evolving curriculum, and 6) an opportunity for alumni and speaker interaction that relays latest discoveries, allows career networking, and provides supplementary evaluation of the training program. The CTP Training Outcomes continue to be outstanding as reflected in published productivity, trainee extramural grants, and job placement (100%-predoc and 100%- postdoc in research-intensive and -related positions). Nine expert chemosensory trainers will shape the intellectual and scientific practice of trainees at two levels as they bridge to independent and externally-funded scientific research programs in chemosensory problems important for the quality of life and human health.