University Of Florida

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Disease-modifying therapies for osteoarthritis (OA) remain a significant clinical challenge. Though numerous promising drug candidates are discovered, they suffer from persistent delivery challenges. The timing and dosing of multiple therapeutics targeting different disease mechanisms must be considered for effective management of OA. Due to issues with joint bioavailability and systemic toxicity, most OA therapies are studied by direct injections into affected joints. Intra-articular injections, however, require image-guidance and specialized personnel, and carry a risk of mis-injection and tissue injury. As such, this approach is limited, especially for repeat and/or timed treatments. Further, in patients with multi-joint disease, these intra-articular injections may not be feasible or tolerable. In contrast, intravenous injections are relatively easier and cheaper to administer, and are less painful and risky. However, to be successful, the drug must cross numerous physiological barriers to localize to diseased OA joints and with minimal off-target or systemic effects. Towards this goal, this proposal will identify peptides that preferentially localize to OA joints after intravenous injection, and leverage these peptides in for the development of targeted OA drug carriers. Aim 1 will use an in vivo phage display to select peptides that home to OA joints from the systemic circulation in an animal model of post-traumatic (PT) OA. In Aim 2, we will attach the selected peptide(s) to nanoparticle drug carriers and evaluate their ability to target to OA joints after intravenous injection. Nanoparticle tracking will be accomplished using conventional optical imaging techniques, and an emerging imaging modality, Magnetic Particle Imaging (MPI) to enable robust and quantitative assessment of nanoparticle biodistribution. Overall, this work will make important advances in systemic delivery systems for OA, which are needed to expand clinical options for comprehensive, long-term therapeutic strategies for OA.

NIH Research Projects · FY 2025 · 2024-07

The overarching objective of the proposed program is to provide an enriched and innovative predoctoral training experience to foster the next generation of stellar neuroscientists. The Broad-based Research, Analytics, and Innovation in Neuroscience (B2RAIN) Predoctoral Training Program takes advantage of the University of Florida’s exceptional strength in neuroscience and its burgeoning Artificial Intelligence/data science communities. Given these strengths, the University of Florida (UF) is uniquely suited to offer a training program that strongly aligns with the stated goals of the Jointly Sponsored NIH Predoctoral Training Program in the Neurosciences and to offer the first such program to predoctoral trainees in the State of Florida. Indeed, UF offers an exceptional environment for the B2RAIN program that includes a strong cadre of dedicated and productive mentors. The B2RAIN program is carefully designed to integrate a well-rounded core neuroscience curriculum with coursework and practical experiences that will enable neuroscience predoctoral trainees to build strong quantitative skills that emphasize AI principles, coding, experimental design, and statistical rigor. Moreover, students will acquire a full complement of critical interdisciplinary skills around communication, collaboration, rigor, and resilience that will bolster individual scientific and professional success. The requested funds, together with a strong institutional commitment, will enable a minimum of eight neuroscience predoctoral students to enter the program annually and for these students to be fully supported during their initial two years of doctoral training while acquiring the essential foundational skills offered by the B2RAIN program. A minimum of forty doctoral neuroscience students will benefit from this training over the course of 5 years. Following the comprehensive training in fundamental neuroscience and rigorous quantitative approaches offered by the B2RAIN program, the students will be well-prepared to complete doctoral research with one of over forty outstanding preceptors. These mentors, who hail from across the UF campus, are dedicated to the training mission and offer a wealth of expertise across a wide range of neuroscience subdisciplines including systems/behavioral neuroscience, neurogenetics, cognitive neuroscience, and neurodegeneration. Trainees will be recruited from a talented cohort of applicants to the UF neuroscience Ph.D. program and will demonstrate the potential to become the next generation of innovators to drive progress in uncovering the complexities of the nervous system in both health and disease. The emphasis on quantitative skills, scientific rigor, and on acquiring essential professional skills important for long-term career success will add significant value to the participating students’ doctoral training experience. Specifically, the B2RAIN Training Program will equip trainees with the cross-disciplinary knowledge, skills, and perspectives to launch and maintain productive research careers at the forefront of the rapidly-advancing neuroscience field.

NIH Research Projects · FY 2024 · 2024-07

Overall Abstract ABSTRACT Florida is a demographically and geographically diverse state. The University of Florida (UF) and Florida State University (FSU) CTSA hub will work within this environment to further the mission to improve human health by accelerating the translation of scientific discoveries and the implementation of evidence-based best practices for the diagnosis, treatment, prevention and cure of human diseases across the lifespan. The UF-FSU hub vision is “think globally, act locally”, offering research opportunities to underserved participants in North and North Central Florida, creating innovative career development and training opportunities, as well as collaborating across the country with the Accelerated Clinical Trials Network and PCORnet. Research strengths include precision medicine, team science, community engagement, implementation science and informatics which are conducted in diverse settings through the OneFlorida Clinical Research Consortium. The 2019-2024 period represents the next phase of evolution from creating a clinical and translational science infrastructure to enhancing the local, state and national impact of CTSI-led science. During this period, FSU will be integrated across all components and will engage six additional colleges. Hub activities will be centered around four strategic goals: (1) chart new pathways for developing the translational workforce by taking UF’s success in career development and translating this success to the FSU and historically black colleges and universities; (2) strengthen the capacity of the learning health system environment and develop transferable models for embedding translational science into the clinical enterprise by further integrating data and software, developing multi-site pilots in healthcare institutions serving unique patient populations and building on the success of the personalized medicine program to use genomics data to improve patient outcomes; (3) expand statewide collaborations and opportunities to advance a participant-centered research agenda that reflects the health priorities and diversity of the catchment area by continuing to strengthen stakeholder engagement and trust in research through the HealthStreet Program and enhance collaboration with policy stakeholders from Florida Medicaid, Florida surgeon general and others; and (4) strengthen regional and national collaborations to accelerate the collective impact of the CTSA network through continued work with ACT, PCORnet, genomic medicine, aging and metabolomics. Throughout this important work, the UF-FSU hub will remain dedicated to supporting the recruitment, retention and career development of underrepresented minority and disabled trainees and faculty. Integral to the success of the proposed work, the UF-FSU hub will further integrate health care and research in Florida guided by four keys to success: (1) organizational alignment, (2) clinical informatics, (3) clinician and stakeholder engagement, and (4) strong support from implementation and improvement science expertise.

NSF Awards · FY 2024 · 2024-07

When thin layers of liquid with a free surface are heated, they create motion called Marangoni flows. These flows occur because of changes in surface tension when the fluid is heated. The Marangoni flows can cause the thin layers to break apart and dry out. This dry-out is detrimental to several materials processing techniques such as optical film deposition and 3D printing. It is also harmful to heat exchangers that are crucial to high computing device performance. Researchers believe that vertical shaking of the thin fluid layers can prevent flows from occurring and therefore stave off dry out. To test this idea, experiments will be conducted in microgravity on the International Space Station to avoid the interference caused by Earth's gravity. Besides scientific discovery this project promotes diversity and inclusion by offering educational opportunities for graduate, undergraduate, and K-12 students. It fosters critical thinking, experimental design, and communication skills through STEM outreach, enriching research with diverse perspectives. The main goal of this research is to determine how and when parametric forcing can prevent Marangoni flows in non-isothermal thin fluid layers and under what conditions they can lead to resonance. The proposed microgravity experiments with multiple test-fluid systems will validate theoretical predictions. Importantly, they will isolate the physics of parametric-forcing dynamics without interference from buoyancy-driven convection. The intellectual merit of the study is that it will provide compelling evidence that there are two regimes of parametric forcing on thermo-capillary flows: a low-frequency regime where such flows can be eliminated and a high-frequency regime where it cannot, but where resonance occurs and where heat transfer can be substantially increased in a closed system. The first regime finds applications in materials processing of thin films and in additive manufacturing. The second regime finds applications in micro-reactors and in micro heat pipes. These applications have substantial broad impacts and benefits to life on Earth, such as in the formation of thin optical films, directed energy deposition, and in the thermal cooling of high-speed computational devices. The study also has broad translational application to the physics of electrostatic and magnetic-induced resonance instability. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT The persistence of latently infected memory CD4+ T cells releasing a low-level trickling of viral RNAs and proteins in people living with HIV (PLWH) on antiretroviral therapy (ART) provides a strong rationale for the development of cure strategies. HIV functional cure approaches are being explored, at the transcriptional level, to achieve sustained ART-free viral remission that would result from epigenetic changes over time. These approaches are supported by evidence in PLWH who spontaneously control their viral load below the limit of detection, in the absence of ART (elite controllers, ECs). These ECs were shown to have a large proportion of their proviruses in condensed chromatin (heterochromatin) loci, with virus refractory to reactivation. The chromatin environment surrounding the HIV promoter is believed to be regulated by host transcription factors, host chromatin remodelers and the viral Tat protein. Our hypothesis is that the pioneer factor (PF) family may also remodel the HIV locus chromatin, via their unique epigenetic modulator properties, resulting in the regulation of HIV transcription. PFs play critical roles in chromatin remodeling, mostly during development and disease phenotypes. Unique properties of PFs include 1) trigger assembly of regulatory factors on target DNA in heterochromatin; 2) bookmark genes for rapid reactivation/repression and 3) induce persistent epigenetic modulation of the cellular chromatin. A few PFs were reported to modulate HIV latency and persistence in primary CD4+ T cells but were poorly studied mechanistically. Preliminary data using shRNAs targeting 13 PFs in the HIV latently infected CD4+ T cells, revealed 3 novel hits that hindered HIV transcription. This data further highlights the significance of uncovering the role played by the PFs on HIV transcription in CD4+ T cells, with the long-term goal to harness them for HIV cure approaches and/or diagnostic purposes. Here, we propose to: Aim 1. Screen a shRNAmir library targeting 31 PFs (and controls) by flow cytometry, taking advantage of the HIV reporter pMorpheus-V5 vector in Jurkat T cells. This vector allows the distinction of productively infected, latently infected, and uninfected cells. The shRNAmir relative abundance in these populations will reflect their activity on HIV transcription. The follow-up of the candidates will be based namely on reproducibility and novelty. Aim 2. Validate the hits identified in preliminary data and Aim 1. Their activity will be confirmed in cell line models of latency with different HIV transcriptional environments, and in primary blood CD4+ T cells from uninfected and virally suppressed ART treated female and male donors since estrogen modulates certain PFs’ activity. Their impact on CD4+ T cell proliferation, activation and cytotoxicity will also be measured. Aim 3. Characterize the 3 hits identified in preliminary data. Their recruitment to the host/HIV promoters (ChIP-Seq analysis), their modulation of the HIV chromatin (MNase nucleosomal mapping, enChIP-MS) and transcriptional regulation (RNA Seq analysis) in cell models of HIV latency will be studied for their comprehensive transcriptional regulatory profiling. Cell based- and in vitro- assays will define their dependence to HIV Tat.

NIH Research Projects · FY 2024 · 2024-07

ABSTRACT With the highest HIV incidence rates observed in the US, Florida strives to develop effective and sustainable HIV prevention strategies. HIV screening and pre-exposure prophylaxis (PrEP) are proven interventions to prevent transmission and reduce new HIV infections. However, uptake of these HIV prevention services is low among women relative to their needs and male counterparts. One of the key barriers to promoting HIV testing and PrEP among women is the challenge of identifying women at risk of HIV acquisition by healthcare providers. Researchers demonstrated potential HIV risk prediction models using electronic health records (EHRs) among men. Unfortunately, they failed to identify HIV risk among women due to having fewer women HIV incident cases in their datasets and a lack of risk factors tailored for women. To fill the gap, we propose to develop an HIV risk prediction model specifically tailored for women and integrate the prediction model into an EHR system as a clinical decision support prototype to assess its feasibility, acceptability, and usability with primary care providers. In Aim 1, we will develop an HIV risk prediction model specific for women to identify potential candidates for HIV testing and PrEP. Leveraging patients’ structured EHRs, ZIP code-linked community-level factors and social determinants of health, and factors extracted from clinical notes via a state- of-the-art natural language processing (NLP) algorithm, we will use AI/machine learning to develop and validate an HIV risk prediction model specifically developed for women (ACTION-HIV algorithm). The results will be used to design a clinical decision support (CDS) prototype to help providers better identify women in need of HIV testing and PrEP. In Aim 2, using a user-centered design approach guided by the five “rights” of the CDS intervention framework, we will conduct 6 focus groups with providers to design and prototype a women-specific HIV risk prediction CDS tool (ACTION-HIV CDS). In Aim 3, using think-aloud protocols and surveys, we will assess the feasibility, acceptability, and usability of the ACTION-HIV algorithm and CDS in a simulated EHR environment with 20 primary care providers at the UF Health outpatient clinics. We will integrate our ACTION-HIV algorithm into UF Health’s EHR system (Epic) to produce the CDS alerts for HIV testing and pilot test ACTION-HIV CDS in a simulated Epic environment presenting synthetic patient data. Our proposed research is highly innovative as it expands past HIV risk prediction models and pioneers in designing and prototyping HIV testing and PrEP-related CDS in primary care settings. This project will provide valuable insights into a future clinical trial which we will investigate the efficacy (including patient outcomes such as rates of HIV testing and rates of PrEP prescription) of a women-specific HIV risk prediction CDS tool within the real-time EHR production in real-world settings.

NSF Awards · FY 2024 · 2024-07

This project aims to serve the national interest by significantly enhancing the spatial communication abilities of students on virtual field trips. Field trips are one of the most common methods used to deliver real life situated learning experiences to students. They represent a form of active learning, enriching traditional lessons by better engaging students and strengthening spatial, verbal, and math skills for many students. However, field trips also present major logistical, financial, and accessibility challenges for many educational institutions. In this IUSE level 3 project, Arizona State University in collaboration with the University of Missouri plans to engage in a 54-month project, the goal of which is to investigate the use of virtual field trips to improve student learning through a virtual field trip platform. The approach leverages web-based digital environments to deliver multi-user, synchronous, situated learning experiences offering students in-depth spatial communication practice. The project features a transformative change in the ways that spatial communication learning is conducted in STEM, democratizing it by making field trip experiences fully accessible anywhere, anytime. Broad dissemination of findings is assured through extensive faculty professional development and the implementation of iVisit in courses, workshops, webinars, and outreach. Intentional efforts will be made to reach low-income and students from groups underrepresented in STEM using iVisit, further improving STEM inclusion. Research questions posited to guide the investigations include (1) What learning affordances in virtual field trips foster active spatial communication and student group engagement? and (2) How do virtual field trips improve student group spatial communication learning? A design-based research approach is to be employed to obtain answers since this offers the opportunity to improve educational practice through iterative analysis, design, and implementation. Researchers and practitioners will collaborate in real-world settings to create and test design principles and solutions for educational core areas in the current curriculum of Construction Management programs in the United States. The iVisit field trip contents and technology will be informed by the conceptual frameworks for situated learning, including Problem-Based Learning (PBL), Computer-Supported Collaborative Learning (CSCL), and spatial communication in construction. Focus group interviews for instructors recruited to provide the field trip experience for their courses, will be recorded, transcribed, and analyzed via thematic analysis. By leveraging the results from the focus groups, the field trip contents and the PBL Student Activities will be created. In order to support the created contents and activities, multiple media representations of spatial data will be embedded into iVisit to digitally embody the required knowledge for learning spatial communication. These field trip contents and PBL student activities will then be used for the iVisit platform development. The NSF IUSE: EDU Program supports research and development projects to improve the effectiveness of STEM education for all students. Through the Engaged Student Learning track, the program supports the creation, exploration, and implementation of promising practices and tools. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

Understanding how specific actions of healthcare settings affect communication between providers and parents will inform the development of tailored or complementary strategies aimed at increasing cancer prevention. Text & Talk is a 30-clinic pragmatic trial in Florida assessing the effectiveness of two implementation strategies: (1) brief provider education and communication training and (2) parent-targeted educational text message reminders. Text & Talk has 58 participating providers (37 trained in communication skills during the study period and 21 to be trained in communication skills after data collection) and 7,837 11- to 12-year-olds randomized for their parents to receive educational text messages about available cancer prevention options. The goal of this proposal is to understand how providers’ integrated the communication training skills from the provider education training into their practice and for which families and geospatial areas the educational text messages were most useful. For the provider-targeted education and communication training, the hypothesis is that providers will have varying experiences with implementing and sustaining the trained communication skills in their practice that will guide improvement and refinement of future education and communication trainings. For the parent-targeted educational text messages, text message effectiveness is hypothesized to be moderated by individual and geospatial factors. Specific aims are to: (1) Evaluate providers’ experiences with and sustainability of integrating communication training skills into their practice; and (2) Measure moderation of the effectiveness of a parent-targeted educational text message intervention by individual and geospatial differences. Hypotheses will be evaluated by triangulating data collected in Text & Talk with additional data collected during the extension period via mixed methods including in-depth, semi-structured provider interviews, provider behavioral surveys about continued use of trained communication skills, and spatially linking addresses. The innovations are a more in-depth focus on provider learning than previous studies and expanding existing understanding of parent-targeted educational text messages. The significance lies in the potential to improve the communication from healthcare providers and practices to parents about cancer prevention opportunities and other important health outcomes.

NSF Awards · FY 2024 · 2024-07

Young children’s language development is crucial for their later academic success. During the preschool years, teachers contribute to children’s language development by engaging in conversations with them. However, it is not clear how often preschool teachers have language interactions with children, or if every child in the class receives opportunities to talk with teachers. This project uses innovative sensing technology tools to examine teachers’ language interactions with preschoolers over the course of a school year. The project employs advanced speech processing algorithms that automatically analyze language data, providing a more detailed understanding of how language is used in the classroom than has previously been possible. The results provide insights into how preschool language experiences shape language development and whether children’s language experiences differ based on their background characteristics, such as their standardized English language skills, temperament, or gender. The eventual goal of this work is to provide teachers with concrete, actionable data on how to enrich language experiences in preschool and lay a strong foundation for future reading success. The study seeks to answer the following research questions: How often do teacher language interactions occur in preschool classrooms? Which children in a classroom are involved in fewer or briefer teacher language interactions, as compared to their peers? Do teacher language interactions change over the course of the year? Are teacher language interactions associated with growth in children’s English language skills? Monthly data are collected in preschool classrooms using sensing technology tools to address these questions. These sensing technology tools allow for the identification of language interactions, or moments when a teacher and child are in proximity to each other and are talking. Information is also collected on children’s language skills, temperament, teacher-child relationships, and demographic information. The outcomes of this project are expected to inform models of language development in preschool-age children and provide fine-grained data that can be used by educators to equitably support language development in preschool settings. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Odors are capable of potently influencing emotional and behavioral responses. The ability to detect odors and assign them hedonic value, or valence, is essential to navigating a world filled with rewards and dangers. Olfactory sensory loss is correlated with changes in emotional responses, reductions in memory, lack of appetite, and overall apathy. Despite strong evidence that odors acquire appetitive and aversive valence, the neural circuitry underlying odor valence learning is unknown. The basolateral amygdala (BLA) acts as a hub for the integration of multisensory information, including olfactory input, with aversive and appetitive outcomes. While there is ample compelling evidence that the BLA encodes for positive (reward) odor valence encoding, whether the same holds true for negative (aversive) odor valence learning is not clear. This is especially true given that studies to date used mildly aversive quinine, which could obscure the firing dynamics associated with more highly aversive outcomes. Dopamine is a potent neurotransmitter that influences cellular plasticity and learning in the BLA, but its influence on odor-valence behavioral learning is unclear. In the proposed F31 NRSA, I will test the hypothesis that through learning, BLA neurons represent the full spectrum of odor valence, both positive and negative, and that the display of learned odor valence behaviors is regulated by dopamine. I will use a combination of electrophysiological and computational methods, optogenetic stimulation, and site-specific pharmacology as mice engage in a go/no-go behavioral task wherein odors are either rewarded or punished. The results from Aim 1 will demonstrate that BLA neurons encode the full spectrum of learned odor valence while the results from Aim 2 will determine the influence of dopamine within the BLA on odor valence learning behavior. Overall, these findings will enhance our understanding of how the brain processes and assigns emotional significance to odors. Furthermore, the results from this project will expand our understanding and appreciation for the role of dopamine in olfactory processing, and has implications for human disorders characterized by disturbed affective regulation.

NSF Awards · FY 2024 · 2024-07

The Sandhills Region of Nebraska is one of the largest intact temperate grasslands in the world. The landscape of the Sandhills consists of grass-covered sand dunes overlaying the Ogallala Aquifer, a vast underground reservoir that supports diverse communities of plants and animals. The most diverse and abundant animals in the Sandhills are the little known and seldom observed microscopic nematodes. Nematodes have adapted to a wide range of Sandhills habitats, from pristine groundwater-fed streams to alkaline lakes with a water chemistry that restricts most other forms of life. It is unknown what allows nematodes to survive in these extreme environments. However, given the combined effects of climate change and groundwater depletion, the Nebraska Sandhills provide a natural experiment to study the forces that drive diversity and adaptations in this ubiquitous group of organisms. This research will provide insights into how nematodes respond to environmental shifts resulting from the changing climate. Graduate and undergraduate students will engage with a multidisciplinary team of researchers to address these topical questions. Of specific interest to this research are freshwater nematodes in three families (i.e., Tobrilidae, Plectidae, and Monhysteridae) representing three evolutionarily distinct lineages known for their physiological and morphological plasticity but also critical to understanding nematode evolution and phylogeny. There are three main research objectives: 1) describing new species from three targeted nematode families, 2) determining their spatial distributions across the Sandhills, and 3) establishing their taxonomic and ecological context. To accomplish these objectives, three Sandhills regions will be sampled including Alkaline Lakes in Year 1, Sheridan and Cherry Counties in Year 2, and the eastern Sandhills in Year 3 for a total of 128 samples. Samples will be collected from a wide range of aquatic habitats such as lakes, fens/wet meadows, and streams/rivers. Each sample will be split into four aliquots and subsequently used for archiving, individual nematode specimen morphological and barcoding analyses, community metabarcoding and mitochondrial metagenomics, and biochemistry. These data will then be analyzed phylogenetically, and other analyses will include species delimitation, phylogenetic endemism, and multiple regression models and co-occurrence networks. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-07

Project Summary The current administrative supplement request is for a 12-month extension with funding to complete the ongoing IGNITE Network pragmatic clinical trials, GUARDD-US and ADOPT PGx. The GUARDD-US and ADOPT PGx trials have been underway since July 2020 and February 2021, respectively. These trials will help determine the impact of implementing genetic testing on hypertension, depression, and pain therapies. GUARDD-US: Chronic kidney disease (CKD) is associated with hypertension. People with African ancestry (AAs) have the highest risk of CKD and kidney failure, the highest prevalence of hypertension, and the lowest rate of blood pressure (BP) control. While this disparity is in part due to social determinants, ancestry has biological underpinnings, and APOL1 high-risk genetic variants, exclusively found in AAs, increase kidney failure risk 10-fold. We propose a genotype-guided trial to determine the effect of early vs. delayed knowledge of a positive APOL1 genotyping result on 3-month systolic blood pressure (SBP). The clinical trial aims to recruit African Americans with hypertension, with or without CKD, randomized to immediate versus delayed return of APOL1 genetic testing. In those who are APOL1 negative, we will also conduct a pilot study to test the impact of pharmacogenetic (PGx) testing on SBP. ADOPT PGx: Pain and depression are conditions that impact substantial proportions of the US population. The treatment of acute and chronic pain is challenged by the difficulty of finding effective therapies while minimizing the risk of adverse effects or opioid addiction. For depression, there are few clinically relevant predictors of successful treatment, which results in inadequate therapy for many patients. We propose a prospective randomized pragmatic genotype-guided clinical trial that tests the effect of genotype-guided therapy in three scenarios of patients: acute post-surgical pain, chronic pain, and depression. For each scenario, participants will be randomized to genotype-guided drug therapy versus usual approaches to drug therapy selection. Changes in patient-reported outcomes representing pain and depression control using standard PROMIS scales define the primary endpoints. Secondary analyses include safety endpoints, changes in overall well-being, and economic impact represented by differences in healthcare utilization. A 12-month extension with funding is needed due to unanticipated network-wide delays in launching each trial and shutdowns due to COVID-19. The funding requested in this administrative supplement reflects the trial close-out needs and the costs associated with leading analyses and publication costs for 13 secondary manuscripts.

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY A critical gap in rotator cuff tear research is understanding why some individuals have a torn rotator cuff but are free of symptoms, while others experience pain and dysfunction. Dr. Pozzi’s Katz R01 (parent project, AR080058) approaches this problem by examining the neural and somatosensory aspects (neuroimaging, central sensitization, psychosocial distress) of symptoms in well-characterized cohorts of symptomatic tears, asymptomatic tears, and healthy controls (no tears, no pain). We have a unique window of opportunity to submit an ancillary project that evaluates the musculoskeletal aspects of symptoms, specifically neuromuscular control and glenohumeral joint health, in the same cohorts as the parent project. The high prevalence of individuals with asymptomatic rotator cuff tears as well as the success of some conservative exercise interventions for symptomatic tears suggests neuromuscular control strategies can mitigate symptoms. However, since the anatomical integrity of the torn tendon is not restored, to what extent previously identified changes of neuromuscular control are adaptive versus pathological remains unknown. Current investigation on the interaction between rotator cuff tears and glenohumeral joint health are limited to murine models, radiographic imaging studies, and studies characterizing end-stage disease (i.e., co-presence of massive tears and glenohumeral osteoarthritis). We will advance this prior work through three aims. In Aim 1, we will elucidate the interrelationship between neuromuscular control and symptom expression in individuals with rotator cuff tears by simultaneously measuring shoulder motion (including scapular motion) and electromyography (EMG) of shoulder muscles (including intramuscular EMG of rotator cuff muscles) during functional tasks. In Aim 2, we will elucidate the interrelationship between neuromuscular control, symptom expression, and shoulder joint health in individuals with rotator cuff tear using static (magnetic resonance imaging to characterize cartilage) and dynamic (musculoskeletal simulations to calculate glenohumeral joint forces) measures. In Aim 3, we will explore potential treatment targets by leveraging recent advances in explainable artificial intelligence to evaluate the relative contributions of various organ systems to rotator cuff symptoms. The organ systems evaluated in the parent (nervous and somatosensory systems) and ancillary (musculoskeletal system) projects will be explicitly studied. Completing this ancillary project will advance our knowledge of the musculoskeletal factors that influence patient outcomes. Integrated analyses of the musculoskeletal (ancillary), neural (parent), and somatosensory (parent) contributions to symptom expression will further elucidate the biological mechanisms driving outcomes, thereby aiding the identification of rotator cuff tear phenotypes and treatment targets.

NIH Research Projects · FY 2026 · 2024-06

Project Summary/Abstract. Chronic musculoskeletal (MSK) pain is experienced by millions in the United States (US), with highest prevalence in older adults, leading to disability. An approach to attain improvement in diagnosis and treatment that has gained momentum over the last decade is to increase understanding of the neural underpinnings of chronic pain by looking at the associated brain changes. Biomarkers of healthy brain aging, derived from machine learning (ML) models that map MRI patterns to chronological age, offer a look at these pathological changes since deviations from healthy brain aging indicate potential underlying pathologies. The Predicted brain Age Difference (PAD; predicted brain age minus chronological age), proposed as a biomarker of disease, was found to be higher with the presence, and positively correlated with the severity, of MSK pain. However, because it is based on a ‘global’ brain age measure, the PAD is limited to signal a ‘poorer health’ state without specifying the type of underlying pathology. This project proposes to develop novel spatially distributed brain age measures (brain age maps) able to capture the brain atrophy signature of different MSK conditions like chronic back pain, osteoarthritis and neck or shoulder pain. These brain age maps will be obtained from T1-weighted MRIs via very innovative convolutional neural network (CNN) architectures that fuse local and global mechanisms of contextual information in the image using the so-called “transformers”. The project then proposes to develop biomarkers specific to these MSK types informed by the brain age maps via CNNs and the so-called “vision transformers”, a cutting-edge methodology ideal for image classification. By accomplishing these goals, this project will reveal useful information about distinct neurobiological mechanisms of different MSK types and their determinants (e.g., aberrant sensory testing or resting functional networks), and how brain age is associated to the multidimensional experience of pain. This could be particularly useful to understand the causes of the high MSK prevalence in older adults. For example, the highest MSK prevalence in older adults might be the consequence of a more ‘natural’ accelerated brain aging, in contrast to more ‘insult-like’ causes in younger adults. Thus, the project also aims to investigate possible age-related differences in PAD maps of MSK. Finally, we evaluate the brain age maps’ ability prognosticate chronic pain chronification and pain-related functional decline. This proposal is powered by the tens of thousands of participants with MRI and pain data in the UK Biobank and leverages the University of Florida’s Artificial Intelligence (AI) Initiative, endowed with one of the most powerful high performance computing infrastructures across universities in the US. With this significant study, the applicant pursues an independent career as an expert in ML/AI methodologies to be used to identify novel pain and brain aging biomarkers. He will also boost his understanding of experimental pain and of the neurobiological mechanisms of pain and aging. He will leverage strong institutional support, stellar mentorship in pain, aging and AI, a thriving collaborative AI network, and powerful computing resources.

NSF Awards · FY 2024 · 2024-06

Modern cyber-physical systems (CPS) are increasingly neuro-symbolic. A typical CPS control pipeline consists of 1) neural networks (NNs), used to process raw high-dimensional data, such as camera images, and 2) downstream symbolic components, such as state estimation and control, that take the NNs' output in order to close the loop. However, there is a fundamental mismatch between the uncertainty on the NN outputs and the assumptions of the downstream components. NNs are known to be vulnerable to even minor input perturbations and distribution shifts that make it hard to characterize the properties of NN outputs. In turn, such robustness issues violate the symbolic tasks' assumptions and guarantees, thus compromising the overall system safety and predictability. The project’s novelties are neuro-symbolic calibration and training methods that aim to repair the fundamental neuro-symbolic mismatch. The project's impacts are safer and more predictable CPS with NN perception across a variety of CPS domains, including transportation, agriculture, and medicine. The research would enable the application of powerful symbolic tasks (e.g., resilient state estimation and robust control) to modern perception-based CPS where the presence of NNs might otherwise violate the symbolic tasks' assumptions. On the educational front, the investigators will co-develop a graduate course on NN calibration that will expose students to the adverse effects of miscalibration in modern CPS and ways to combat it. The main innovation of this project is the formalizing of the connection between calibration, training and neuro-symbolic methods, especially in the CPS domain. There is a need for a CPS calibration framework that: 1) is robust to data artifacts such as temporary sensor faults; 2) provides calibrated outputs that are consistent with system dynamics over time; and 3) considers the assumptions of the downstream symbolic task. We provide a general framework for combining standard (neural) calibration losses with symbolic losses that aims to align the NN outputs with the assumptions of the downstream symbolic tasks. The research agenda consists of two directions: (i) extrinsic neuro-symbolic calibration to align the uncertainty in NNs with the subsequent symbolic tasks without retraining the NNs, and (ii) intrinsic neuro-symbolic training and calibration to simultaneously train and calibrate NNs for the subsequent symbolic tasks. Both directions are being applied to two broad classes of symbolic tasks, namely state estimation and control, for two general types of symbolic assumptions, i.e., probabilistic and bounded inputs. The benefits of neuro-symbolic calibration and training are being demonstrated on a 1/10-scale autonomous racing platform - the F1/10 car. In addition, the PIs will conduct outreach activities within Rensselaer Center for Open Source and Course-Based Undergraduate Research Experience (CURE) with the Center for Undergraduate Research at the University of Florida. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-06

The overall goal of this project is to better understand how the nervous system evolves. By linking traditionally separate fields of genetics and neuroscience, the researchers will reveal how sensory systems change over time in the blind Mexican cavefish. The cavefish is a special animal in that it has both blind and eyed populations, thereby providing a unique opportunity to connect basic, fundamental research with a broader understanding of evolution for society (through journal publications, textbook chapters, popular press coverage etc.). The results of this work will generate new techniques and ideas that can be used to better understand how genetic variation contributes to neural processing in naturally evolved populations. In addition, this work on how the hair cell system processes information will provide insight into human auditory and vestibular (sense of balance) systems, as well as inspire engineers seeking to design efficient, low-cost sensors in autonomous vehicles. Furthermore, the researchers will present their work to the thousands of K-12 students who participate in the Whitney Lab’s Scientist for a Day Program, enhancing scientific literacy in Florida. New content on “sensory brains” will be developed for the YouTube channel, Fish Code Studios (established in 2014), that reaches a global audience, with 11,700+ subscribers and over 450,000 views and will continue to serve as a major public outreach platform. Researchers will investigate how selection influences sensory detection and modulation by applying powerful physiological and genetic approaches to an emerging model system for neural evolution. Combining electrophysiology and genome wide analyses of genetic architecture, investigations will reveal how the cavefish lateral line system evolved to enable prey capture. Aim 1 will focus on neurophysiology underlying variation in lateral line sensitivity across development. The lateral line system in fishes consists of mechanosensitive neuromasts distributed across the body, with the hair cells of each neuromast connected to afferent neurons that relay information to the brain. This information is simultaneously regulated by an efferent feedback circuit to enhance flow signals during swimming. Using in vivo patch clamp recordings, lateral line sensitivity in surface and cavefish populations will be defined to test whether evolutionarily derived changes in lateral line physiology have enhanced the sensory capabilities of cavefish. This will be done for the first time across developmental stages in larval cavefish. Aim 2 will involve a comprehensive genome wide analysis of genetic architecture regulating lateral line-mediated prey capture behavior. To identify the genetic architecture associated with lateral line differences, F2 surface-cave hybrid fish will be generated and the relationships between quantitative genetics and behavior will be measured. Genotyping-by-sequencing and high-resolution quantitative trait locus (QTL) mapping will be used to identify genomic regions associated with lateral line function and prey-capture behavior. This aim will determine the relationship between lateral line genetics and biomechanical differences in behavior. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-06

Project summary JC virus (JCV) is a human polyomavirus which infects most of the human population worldwide during early childhood with 50-70% of the adult population being seropositive for the virus. JCV is thought to be transmitted by the oral/fecal route however the primary site of infection has not been identified. Following an initial infection, JCV transits into the kidneys where it establishes a lifelong infection. For most people this infection is latent and no new virus particles are detected, however about 8-20% of people have a low-level virus production leading to the excretion of virus into the urine. For immune-competent individuals these lifelong infections will be asymptomatic. However, for immune-suppressed patients, these viruses can reactivate leading to fatal disorders. Upon immunosuppression, JCV will leave the kidney and move to the brain where it will infect the myelin-producing oligodendrocytes. Infection of the oligodendrocytes will lead to their lytic destruction causing the fatal disorder Progressive Multifocal Leukoencephalopathy (PML). JCV genomes have been found in both upper and lower gastrointestinal tissues and in feces suggesting that human intestinal cells can support JCV infection. While the consensus view is that initial infection of the human population by JCV is by ingestion, direct evidence of infection of the intestine is lacking. Most importantly, how JCV passes the intestinal epithelial cell barrier en route to the kidney remains unknown. We plan to fill these gaps in knowledge and study how JCV infects, replicates, and spreads in human intestinal epithelial cells. Understanding the enteric phase of JCV is of major significance to understand JCV pathogenesis as it is believed to constitute the entry site from which 70% of the population becomes infected. In a set of preliminary data using human intestinal organoids, as a surrogate system to study JCV infection in human primary intestinal cells, we could show that JCV can replicate in human intestinal cells and produce de novo infectious virus particles. In this proposal, we will build on our preliminary data and address the cellular tropism of JCV by evaluating the susceptibility of different human intestinal cell types to JCV infection. Additionally, we will test whether all sections of the gastrointestinal tract (duodenum, jejunum, ileum, and colon) can support JCV infection. Finally, we will determine how JCV passes the intestinal epithelial barrier en route to the kidney to establish its chronic infection. We anticipate that this work will provide us with unique opportunities to identify novel therapeutic to prevent infection/transmission from the intestinal mucosa. Importantly this work, will constitute the building block for future research that will integrate the natural gut/kidney axis that is likely to play an important role in JCV infection and pathogenesis.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Subacromial pain, a generic term used to describe pain with disorders of structures within the subacromial space, is the most common reason for shoulder-related visits to primary care and has a high one-year prevalence. Patients with subacromial pain frequently complain of difficulty using the arm, impaired lifting, and limited exercise tolerance. Strengthening is a clinically relevant outcome in patients with subacromial pain. However, the high loads required for strengthening exacerbate pain and stress the rotator cuff tendons. Therefore, clinical practice guidelines recommend strengthening with low intensity and high frequency, but this combination often results in undertraining exercise prescriptions and lasting weakness. Blood flow restriction training (BFRT) is emerging as a novel adjunct treatment to address muscle weakness. It consists of strengthening exercises while an inflatable cuff restricts blood flow at the extremity. It is popular with athletes because it promotes muscle hypertrophy. It is also highly appealing as a rehabilitation strategy because it requires low external loads for strengthening. BFRT could have a tremendous positive impact on the recovery of patients with subacromial pain, but research is lacking. Our long-term goal is to conduct a large-scale efficacy trial of BFRT in patients with subacromial pain. As a critical step toward our long-term goal, this proposal is a pilot randomized trial to test trial procedures, safety and feasibility of the intervention, and preliminary efficacy of BFRT. Our pilot trial will be double masked (investigator & outcome assessor), include two parallel arms (active or sham BFRT), and two endpoints (after 16 BFRT sessions, primary endpoint; and after six months, secondary endpoint). This design is robust because it controls the non-specific effects of receiving BFRT (sham). The first aim tests recruitment, randomization, participant masking, retention, and data collection procedures. The second aim determine the safety and feasibility of the interventions (active BFRT and sham BFRT). The third aim describe the effect of active BFRT on clinical outcomes (shoulder strength, patient-reported outcomes, shoulder performance, muscle hypertrophy, and tendon thickness & elasticity). This application is innovative because: a) applies BFRT in a clinical population with subacromial pain; b) has clear objectives, analytic plans, and criteria to establish the success of the trial procedures, safety, and feasibility outcomes; and c) uses comprehensive outcomes to evaluate clinical recovery. The popularity of BFRT among professional athletes favors quick transitions to clinical applications even without support from high-quality research. Completing this preliminary trial will demonstrate that we have the capacity and capability to conduct and deliver a large-scale efficacy trial as planned. Collectively, this line of research will directly impact clinical practice.

NSF Awards · FY 2024 · 2024-06

With the proliferation of high-tech gadgets such as smartphones, tablets, wearables, and smart-home devices, the amount of data that needs to be made secure is increasing. Malicious attackers target even encrypted devices to steal confidential information using advanced techniques. Side-channel attacks, which exploit various sources of leakage such as power consumption, timing data, and electromagnetic (EM) emissions from a computing device during data handling processes, are commonly utilized to breach the security of integrated circuits (ICs). Due to their cost-effectiveness and potentially non-intrusive methods, these attacks pose a significant threat to the confidentiality of personal, commercial, and military data. This project leverages the synergetic collaborative effort between the USA and Indian research teams to provide security against power and EM side-channel attacks at different abstraction levels, including EM analysis, circuit/logic/layout design, and architectural/system-level choices. The project develops secure libraries and tools that can be used by circuit design engineers and a scientific methodology to evaluate EM and power side-channels in custom ICs. The outcome of this project makes a positive impact on the electronics industry through security-aware design, pre-Silicon side-channel vulnerability analysis, and quantifying security using commercial design tools. This project also provides integrative, multidisciplinary training to graduate and undergraduate students on a multitude of technical areas such as security-aware circuit, layout and package design, development of custom tool flows, and characterization/quantification of security on hardware devices. This project also fosters increased collaboration between USA and Indian researchers, potentially setting the foundations for long-term engagement and scientific partnerships. The project team's technical approach entails a massive co-design of electromagnetics, materials, circuits and architecture for secure IC design. To this end, this USA team conducts the research on modeling of the EM and power side channels for various real-world devices, along with the design and analysis of on-chip side-channel countermeasures. The Indian research team leads the pre-silicon side-channel security analysis for EM and power, as well as standard cell library design for security enhancement. The design of the side-channel secure standard-cell library utilizes a novel split-pole based CMOS design methodology that creates complementary dipoles to cancel EM emanation at the gate level. This effort is complemented with side-channel aware on-chip global routes utilizing dipole, quadrupole, and higher order EM emission characteristics, which helps in reducing EM side-channel leakage resulting from long on-chip metal lines. Designing encryption cores with the side-channel secure standard cell library is further benefitted from a configurable, design-tool-friendly version of a custom current-domain signature attenuation circuit. The research team is also exploring design-tool-aware and timing-aware clock path optimization techniques to reduce the EM and power side-channels. Additionally, material properties of metal layers in CMOS ICs are being analyzed to provide potentially better side-channel resistance for both IC-level and three-dimensional (3D) integrated package-level implementations. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2024-06

Chronic musculoskeletal conditions and their primary source of pain, movement-evoked pain (MEP), causes significant pain interference, long-term mobility impairment, healthcare costs, and psychosocial inequalities for individuals, families, communities, and society-at-large. MEP affects more adults in later life and is perceived as more severe than resting pain. Many older adults hesitate to use opioids and non-steroidal antiinflammatory drugs long-term and rely on non-pharmacological pain self-management (SM) strategies. Current studies of chronic musculoskeletal pain (CMP) SM interventions do not measure MEP as a primary endpoint nor address faith/spirituality, varied traditions, and underlying potentially modifiable health indicators (e.g., health literacy, financial hardship), and therefore only offer small or no benefit over control conditions. We propose to enhance older adults’ capacity to manage MEP by investigating the effects of a tailored pain SM intervention (Pain Relief for musculOskeletal conditions and Arthritis using Carefully Tailored InterVEntions [PROACTIVE]). This prototype intervention will provide focused pain SM education with a SM resource toolkit, active prayer, and financial counseling to explain participants’ healthcare benefits for pain care. This intervention moves beyond basic tailoring and targets multiple areas for behavior change that are of importance to adults in later life. We have assembled an interdisciplinary and clinically experienced team to lead this study. We will enroll 120 adults (age 50-92 years) with CMP into a parallel group, single-blind, randomized controlled trial to test whether PROACTIVE decreases MEP (primary outcome) in the immediate post-intervention period (n= 60) (Aim 1) and over time (Sub-Aim 1a) compared to a usual care control group (n= 60). To determine efficacy, we will use state-of-the-art and real-time technologies to measure MEP as well as pain interference, pain coping, and physical function (secondary outcomes) (Sub-Aim 1b). The PROACTIVE group will work with a nurse and financial counselor over four weeks to enhance knowledge of CMP SM, utilization of active prayer and faith, and financial literacy of health insurance benefits and resources available to cover treatments for pain. Each weekly session will last up to 2 hours and will be followed by six days of ecological momentary assessments of pain and SM outcomes and ecological momentary interventions guiding participants through weekly SM skills. A fourth booster session will reinforce content and training and help sustain SM. Also, we will examine the differential effects of PROACTIVE on MEP according to demographic and psychosocial factors (Aim 2). The proposed intervention is expected to produce meaningful reductions in MEP in aging adults experiencing disabling chronic musculoskeletal conditions. Ideally, this study will identify precision behavior targets and responders to inform intervention refinement.

NSF Awards · FY 2024 · 2024-06

This grant will support travel of students and post-doctoral researchers who are US citizens or US permanent residents to attend the Hilton Head Workshop 2024 (HH2024): A Solid-State Sensors, Actuators, and Microsystems Workshop. Specifically, this grant enables the HH2024 organizing committee to support participation of diverse attendees with a focus on those underrepresented. The committee will select the awardees from the pool of high school, undergraduate and graduate students, and postdoctoral researchers who are the first or second authors of the accepted papers. HH2024 organizing committee will target awarding approximately 50 percent of the awards to women or underrepresented minorities. The 21st in the series of Hilton Head Workshops (HH2024) on the science and technology of Solid-State Sensors, Actuators, and Microsystems will take place 2-6 June 2024 at the Sonesta Resort on Hilton Head Island, South Carolina. This exciting multidisciplinary event has occurred biennially since 1984, and this year the Hilton Head Workshop’s 40th anniversary will celebrate Micro-Electro-Mechanical Systems (MEMS) and microsystems successes over the past four decades. Additionally, participants will look ahead to the challenges and opportunities for the field to contribute to addressing national and global grand challenges in the next 40 years. The HH2024 Workshop is the prominent and top-tier conference in the field of microelectromechanical systems (MEMS) and Microsystems and related fields, and is expected to draw 350-500 academic, industry, and government participants from diverse engineering and scientific backgrounds, including chemistry, materials science, chemical engineering, electrical engineering, mechanical engineering, physics, biology, and bioengineering. This year’s workshop will have a focus on design, fabrication and manufacturing of emerging nano/microscale device and systems for sensing, actuation, and computing with broad impact on biomedical, energy, information processing and quantum technologies. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2024-06

ABSTRACT/ SUMMARY This R01 study will apply neurocognitive, brain, and plasma biomarkers to quantify prodromal signs of dementia in autistic adults ages 40-65. Autism spectrum disorder (ASD) is a lifelong condition that continuously demands precision diagnosis, targeted treatment, and high-quality care. Emerging clinical and epidemiological studies suggest that autistic adults are more vulnerable to developing early-onset dementia than the general population as they age. Robust pilot data from our group show accelerated verbal memory and executive function deterioration in middle-aged autistic adults. Our free-water diffusion MRI (FWdMRI) data show FW accumulations in the hippocampal and frontotemporal areas relevant to these behavioral deficits in ASD. Our amyloid PET scans highlight early amyloid load in Phase 1 (i.e., neocortices) Thal regions and elevated amyloid burden in Phase 2 (e.g., entorhinal cortex, hippocampus, and amygdala) Thal regions in autistic adults, accompanied with cortical thinning and volume reductions of these affected areas. Our preliminary plasma biomarkers also show significant increases in amyloid β42 (Aβ42), Aβ40, total Tau, and neurofilament light chain (NfL) in autistic adults relative to controls. Despite promising findings from our lab and others, studies focused on neurodegenerative comorbidities in ASD are scant. We are still far from a systematic understanding of dementia onset, manifestation, and progression in ASD. To address this critical scientific gap, we plan to acquire cross-sectional data to quantify established neurocognitive, brain, and plasma biomarkers of dementia in autistic adults and compare these markers with age- and sex-matched cognitively normal controls. Forty-five autistic adults and 45 controls ages 40-65 will be recruited. Aim 1 will quantify verbal memory retention and executive function impairments in ASD. Aim 2 will quantify dementia-related brain degenerations in autistic adults using T1 weighted MRI, FWdMRI, and amyloid PET. Regions of interest will focus on the hippocampus, hippocampal input and output tracts, and frontotemporal areas. Aim 3 will focus on quantifying plasma Amyloid/Tau/Neurodegeneration (A/T/N) biomarkers in autistic adults, including Aβ40, Aβ42, Aβ42/Aβ40 ratio, phosphorylated Tau (pTau), NfL, and glial fibrillary acidic protein (GFAP). The relationships among cognitive (Aim1), brain (Aim2), and plasma (Aim 3) biomarkers will be explored to achieve a multi-dimensional evaluation of dementia pathophysiology in ASD. This work holds significant promise to develop a foundational understanding of the pathophysiological mechanisms underlying dementia in ASD across outcome measures at multiple levels.

NIH Research Projects · FY 2026 · 2024-06

Despite advances in our understanding of HIV pathogenesis, a knowledge gap remains concerning HIV viral reservoirs. Unknowns include, their various locations, size of the reservoir and the spatial/temporal kinetics for reservoir establishment. The difficulty in quantifying and mapping HIV using common histological techniques has limited progress towards answering these questions. These challenges are recognized under RFA-DA-23- 001 in a call to action to “develop or exploit in situ imaging technologies to investigate HIV infection, or latent HIV/SIV reservoirs” whilst also in the context of substance abuse. Here we address the above by leveraging the latest in optical tissue clearing and volumetric imaging to resolve single cell information while maintaining the 3D structural arrangement in whole tissue. These techniques provide high spatial resolution that in-vivo imaging cannot match, affecting the ability to identify reservoirs that are sparse throughout the entire tissue. Importantly, this application will focus on novel CNS associated reservoirs including the calvarial bone marrow environment and CSF draining superior lymph nodes. Comparisons will be made to brain viral reservoirs within the basal ganglia and hypothalamus. The overall hypothesis is: Volumetric deep tissue microscopy and machine learning image analysis reveals the level of HIV present, spatial distribution and cells infected within novel CNS-associated sanctuary/reservoir sites. In the R61 development phase, Clarity will be performed to eliminate cellular lipids and generate a physical structural support via transparent acrylamide hydrogels that allows light to penetrate deeply for 3D reconstruction. Due to the inherent variability of organs and species-specific tissue composition, no universal protocol is appropriate. Thus, hydrogel formulations, mechanisms for lipid removal, and analytical determinations for optical clearing will be evaluated. The goal is to analytically determine parameters ideal for excellent structural retention and minimal tissue damage/protein loss in non-human primate whole tissue. Additionally, these clearing methods will be optimized for compatibility with antibody immunostaining and in-situ hybridization of HIV RNA/DNA probes in CNS associated HIV reservoirs. Our evaluations will include evidence for a novel HIV reservoir, the calvarial bone marrow (Aim 1), the superior cervical lymph node (the end site of CSF drainage) (Aim 2) and key areas within the brain (Aim 3). The significance of these studies is reflected on the fact that to date, no published work exists in whole tissue optical clearing of the aforementioned viral reservoirs in non-human primates. In the R33 application phase, protocols developed in the R61 phase will be utilized to determine standing questions about HIV reservoirs such as how soon the reservoir is established, which cell types are infected and to what degree do these sites harbor HIV. The above questions will be answered as a function of chronic opioid exposure (Aim 4) or presence of ART (Aim 5). Overall, these studies will provide critical information on CNS associated HIV reservoirs using novel in-situ imaging methodologies, fulfilling the main objective of RFA-DA-23-001.

NIH Research Projects · FY 2026 · 2024-06

Project Summary/Abstract Feeding is an essential behavior for animals to obtain nutrients for survival, but the influx of nutrients during and after feeding must be properly managed to avoid disruption of metabolic homeostasis. Neural circuits in the brain utilize sensory cues associated with food consumption, such as the taste of food and the amount of ingested food in the digestive tract, to prime endocrine systems for incoming nutrients and regulate feeding pace and amount. Although progress has been made in recent years to identify neural populations that regulate endocrine function and feeding behavior, the complete neural circuits that transform food sensory signals into endocrine regulation and feeding behavioral control are largely obscure. The long-term goal of the proposed research is to elucidate the complete neural circuits that carry out such sensory-endocrine/feeding transformations, and obtain a comprehensive circuit-level understanding of endocrine and feeding regulation and its function in health and disease. To achieve this goal, the fruit fly, Drosophila melanogaster, is used as a model system. Drosophila has long been a valuable model for neural circuit studies, providing insights into common principles for chemosensory processing and feeding regulation that are shared with mammals. The recent completion of an electron microscope connectome of the entire Drosophila brain at synaptic resolution provides an unprecedented opportunity to delineate complete neural circuits linking food sensory detection to endocrine and feeding control. Guided by our connectomic analyses, this proposal aims to test the central hypothesis that a subset of serotonergic neurons in the Drosophila brain integrates both external and internal food sensory signals to regulate endocrine function and the feeding motor program. Aim 1 will use state-of-the- art optogenetic and functional imaging approaches to test the hypothesis that serotonergic neurons integrate food-related gustatory and mechanosensory signals from multiple organs, including external and internal mouthparts and the digestive tract. Aim 2 will test the hypothesis that serotonergic neurons activate two parallel output pathways to (a) regulate endocrine function to promote sugar metabolism and digestive function, and (b) inhibit the feeding motor program to suppress food intake, using a combination of calcium imaging, optogenetic, metabolic, and behavioral approaches. These studies will reveal a complete neural circuit from food sensory inputs to endocrine/motor outputs, and provide mechanistic insights into the sensory integration of external chemosensation and gut interoception, gut-brain-endocrine interactions, and feeding behavior regulation. The findings from this research will advance our understanding of how neural circuits regulate feeding behavior and feeding-related endocrine function, providing a foundation for examining how compromised function of these circuits may contribute to endocrine dysfunction, eating disorders, and obesity.

NIH Research Projects · FY 2026 · 2024-05

This study aims to investigate the dynamic relationship between TDP-43 and Tau proteins in normal and pathological contexts. We will characterize the interaction between TDP-43 and Tau by probing their binding using NMR and surface plasmon resonance. We will also test small molecules and peptides for their efficacy in mutilating or disrupting TDP-43/Tau interaction. Additionally, our study aims to determine the physiological and pathological consequences of TDP-43 and Tau interaction by examining phenotypes, behaviors, and cellular interactions using a humanized knock-in disease model in which normal aging and axonal injury will serve as the paradigm for examining lifespan changes that do not involve genetic overexpression or pharmacological initiation. The identification of binding partners and the role played by phosphorylation within the TDP-43/Tau protein-protein network will further expand our understanding of neurodegenerative disease development. The findings from this study will contribute to the development of novel therapeutics approaches targeting the interaction between TDP-43 and Tau spawning generation first-in-class therapeutics for AD and other neurodegenerative diseases.