University Of Rochester

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY The inner-ear fluids, unlike other body fluids, are stationary and isolated from the rest of the body. These characteristics give opportunities and challenges in maintaining inner-ear health and in treating inner- ear diseases. Due to the blood-labyrinth barrier, systemic delivery of drugs to the inner ear is highly inefficient. On the other hand, this isolation is an opportunity—drugs can be delivered locally with minimal off-target concerns. Unfortunately, the potential advantage of local delivery has been difficult to capitalize on because of the labyrinthine geometry of the inner ear. Application of drug at any location of the inner ear labyrinth filled with stationary fluids results in high concentration at the application site without reaching distant locations. A current remedy is to create surgical holes in the temporal bone to allow inner-ear fluids to flow despite the risk of surgical damage. We propose minimally invasive and efficient drug delivery mechanism into the inner ear. Specifically, we will develop a method to use sounds as the agitating source for cochlear drug delivery. Recent data regarding OoC micromechanics are both exciting and controversial because new observations do not fit well into existing frameworks for cochlear biophysics. For example, the outer hair cells are widely-acknowledged as the actuator for cochlear amplification. However, the outer hair cells generate force most efficiently at frequencies below the characteristic frequency at most cochlear locations, raising the possibility of additional functions. The proposed project combines two topics that have previously been investigated independently—mechanics and fluid homeostasis of the OoC. By combining these two subjects, we propose the novel hypothesis that active outer hair cells enhance mass transport along the cochlea. We will test the hypothesis with three aims that combine physiological and computational modeling approaches. For Aim 1, experiments in live animals (gerbil) will be used to characterize the effect of sound and outer-hair-cell motility on mass (neurotoxin) transport along the length of the cochlear duct. Aim 2 experiments will use excised cochlear tissues implanted in a novel micro-fluidic chamber to characterize the OoC peristaltic vibrations due to outer-hair-cell motility. For Aim 3, new biophysical computer models will simulate drug delivery along the cochlea, thereby integrating physiological results from Aims 1 and 2. Approximately one out of five adults in the United States has some degree of hearing loss. Multiple common forms of hereditary, age-related, and noise-induced hearing loss are ascribed to malfunctions of cochlear-fluid homeostasis. By investigating cochlear-fluid homeostasis from an innovative point of view (mechanics), this project will provide an explanation on why hearing of high frequency sound is more vulnerable. In the long term, we have ambition to provide a remedy to delay/prevent hearing losses related to fluid-homeostasis.

NIH Research Projects · FY 2024 · 2022-09

The outer blood retina barrier (oBRB) comprises of the retinal pigment epithelium (RPE) cells and underlying fenestrated choriocapillaris (CC) that interfaces with the blood supply. The RPE-CC complex functions synergistically to support photoreceptor cell health that is critical for vision. Consistently, dysfunction of the RPE- CC leads to retinal degeneration in myraid eye diseases, including age-related macular degeneration (AMD), the single biggest cause of irreversible blindness in adults > 50 years of age in the US. However, the lack of in vitro tissue mimetics that faithfully recapitulate the RPE-CC complex has significantly impaired the study of normal and diseased physiology of the oBRB. A major challenge for the development of RPE-CC tissue mimetics is our limited understanding of human retinogenesis. This is especially relevant to the CC layer in which the majority of inferences are drawn from histological studies of embryonic human retina. Human induced pluripotent stem cells (hiPSCs) provide a unique platform to develop in vitro oBRB models. Indeed, several studies have now shown that specific cell types relevant to the RPE-CC complex, including RPE, endothelial cells (ECs) and mesenchymal stem cells (MSCs) can be differentiated from hiPSCs. Furthermore, we have recently developed a primitive RPE-CC tissue mimetic by exploiting the versatility of poly(ethylene glycol)(PEG) hydrogel-based engineered ECM (eECM) and hiPSC-derived target cells to emulate the spatial organization of RPE, ECs, and mesenchyme. The RPE-CC tissue mimetic is able to recapitulate important physiological features of the in vivo RPE-CC complex, such as CC-like fenestrated vasculature, that had previously been elusive in vitro. Although this model provides a framework for physiological RPE-CC development, it currently has several limitations, including unoptimized eECM biochemical and biophysical cues, lack of developmentally-instructed temporal cell- cell cues, and absence of vascular perfusion, resulting in a tissue model that does not fully recapitulate in vivo structure (e.g., well-defined Bruch’s membrane-like ECM and CC spatial angioarchitecture) and function (e.g., nutrient transport and macromolecular diffusion). In this proposal, we hypothesize that better understanding of the eECM requirements (Aim 1), ii) incorporation of temporal developmental cues (Aim 2), and integration of vascular perfusion, (Aim 3) will promote development of modular, spatially relevant, and functional RPE-CC tissue mimetic(s). Ultimately, the development of a physiological and modular human outer retina (RPE-CC) tissue mimetic will have important implications for subsequent disease modeling, drug screening, and transplantation studies.

NIH Research Projects · FY 2023 · 2022-09

Project Summary Substance User Disorders (SUDS) and Intimate partner violence (IPV) are devastating to families and society (CT CASE, 2015) costing $700 billion annually in healthcare expenditures for SUDS and $12.6 billion in annual costs for IPV. There is a high co-occurrence of substance use and IPV (IPV; CT CASE, 2015). Rates of SUDS and IPV increased during the Covid-19 pandemic (Dubey et al., 2020) at a time period when access to care was disrupted in the absence of Telehealth or Digital Therapy Platforms. The alarmingly higher rates of SUDS and IPV during the pandemic underscored the need for more clinical research trials for device and digital technology developments (DTx). CBT is an evidence based therapy vehicle that has been shown to be effective in improving treatment outcomes across a number of behavioral health disorders (Dutra et al., 2008) including initial efficacy in treating substance abuse and IPV among male offenders in an individual (1:1) CBT therapy modality (Easton et al., 2017). Recent meta-analytic reviews report that digitized versions of CBT are showing effectiveness in treating a range of maladaptive behaviors (Spek et al., 2007) as well as SUDS (Carroll and colleagues, 2014; Carroll et al., 2008) but are limited digital versions that lack personalization and relevant content to patients self-reported symptom distress. To date, digitized platforms have not been used to treat SUDS and IPV among clients entering substance abuse treatment. Researchers are calling for more RCTs using DTx’s of CBT as a vehicle for healthy behavior change among SUD - IPV clients (Nesset and colleagues, 2019). Given our prior success with CBT to treat both SUDS and IPV across 12 weeks of 1:1 treatment, we extended our integrated CBT therapy to a DTx platform. DTx platforms are advantageous because they are easy to disseminate, cost-effective, lead to increases in clients’ engagement and maximize compliance with practice exercises. The technology that exists today is modernized to allow for personalized therapy content to be linked to reported symptom distress. We developed a 12- week digital, Avatar Assisted, interactive platform, RITch®CBT, as an intervention platform self -guided by patients and for patients to use “at home” to practice coping skill exercises at their convenience. In response to NIH’s PAR 21-183, we propose to conduct a Phase I and II Study: UG3 (Phase I) and UH3 (Phase II) in collaboration with the FDA regarding ongoing feedback and regulatory processes. In Phase I, we propose a feasibility study, a randomized controlled trial to test the efficacy of RITch®CBT among SUD-IPV clients entering addiction treatment comparing it to face to face 1:1 CBT. If efficacy is achieved, an effectiveness study will be performed in Phase II (UH3) to improve treatment outcomes among individuals and their families suffering from co-occurring SUDS and IPV, a common co-occurring problem within families across the U.S.

NIH Research Projects · FY 2026 · 2022-09

PROJECT SUMMARY Chronic pain is still a clinical diagnosis based on location, symptom report, and clinical expertise. Despite recent efforts to delineate specific and evidence-based criteria to diagnose different chronic pain conditions, substantial heterogeneity persists among chronic pain patients often within the same clinical pain syndrome (e.g., low-back pain). The lack of quantitative and reliable measures to diagnose chronic pain and the related heterogeneity that ensues are major obstacles to medical care for patients and for research studies. Chronic pain patients are often managed using a “trial and error” approach as targeted and precise treatment is not possible without quantitative biomarkers, like glucose levels for diabetes. In addition, patient-related variability in analgesic response is thought to be one of the main reasons why the current therapeutic interventions for chronic pain are unsatisfactory, as 20% of US adults live in chronic pain and 8% of US adults are disabled from chronic pain. Natural language processing analyzes semantic and emotional content, syntactic structure, and complexity of speech; audio-visual processing analyzes voice acoustics and facial expressions. These tools have recently been shown to be powerful quantitative and reliable biomarkers for discriminating between patients with psychiatric conditions like schizophrenia and major depression, and in predicting long-term outcomes, like the development of psychosis in high-risk groups. A parallel can be drawn between chronic pain and chronic mental illness like major depressive disorder, as both conditions are diagnosed based on subjective report of symptoms, diagnostic criteria, and clinical expertise. In addition, both conditions are closely associated with negative affect which has been corroborated by preclinical research and brain imaging data showing a critical role of the limbic brain in the pathophysiology of these conditions. Therefore, it stands to reason that natural language and audio- visual processing may serve as biomarkers to phenotype different types of chronic pain patients and to measure patients' responses to treatment. This proposal will study the ability of language analysis and audio-visual processing tools in discriminating between different types of patients with chronic pain (i.e., discriminant validity) in Aim1, and the ability of these tools to predict analgesic response of chronic low-back pain (CLBP) patients receiving spinal cord stimulation (SCS) (i.e., predictive validity) in Aim 2. In both aims patients will be video recorded during an interview where they speak about their pain or mood (for major depressive disorder patients). Language, speech, and facial expression features will be extracted from the recordings and used in multivariate machine learning models. In Aim 1 natural language and audio-visual processing patterns will be compared between patients with 3 conditions: (1) musculoskeletal CLBP, (2) musculoskeletal CLBP with clinically significant negative affect, and (3) moderate major depressive disorder. In Aim 2, natural language and audio-visual processing patterns will be used to identify responders and non-responders to SCS.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Rheumatoid arthritis (RA) represents a chronic progressive process which leads to significant morbidity and mortality. RA is driven by both inflammatory and stromal pathologies, and while current therapies improve inflammation, there are not effective treatments targeting fibroblasts and bone pathology in RA. The CD47 pathway can affect both immune cell phagocytosis through SIRP-a signaling and stromal pathology through TSP-1 signaling. This study will define the role of CD47 signaling in patient biospecimens and mouse models of arthritis, and assess the utility of combinations of anti-CD47 therapy and biologics in RA. Our central hypothesis is that CD47 is critical to RA pathogenesis and that its blockade will ameliorate or reverse inflammatory arthritis and bone erosion. We will test this hypothesis through three specific aims. Aim 1 will characterize the role of the CD47 signaling through TSP-1 and SIRP-a in patients with RA through histologic analysis, single cell RNA sequencing, and synovial organoid cultures. Aim 2 will assess whether CD47 is required for inflammatory arthritis by assessing arthritis, bone outcomes, and cellular function in mice deficient in CD47 after inducing arthritis. Aim 3 will determine the effectiveness of CD47 inhibition in combination with biologic therapies in treating arthritis first using an in vitro drug screen and then testing the most promising candidate therapy in vivo. The proposed research is significant both because it will substantially improve understanding of RA biology, and because it has the potential to identify novel therapeutic strategies which can treat both inflammatory and stromal pathways in RA.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Numerous genetic variants in tRNA modification enzymes have been linked to devastating neurodevelopmental and neurological disorders. However, the molecular mechanisms underpinning these pathologies are unknown. Why is it that perturbations to many different tRNA modification enzymes and thus, changes to the various chemical modifications they catalyze, seem to affect the brain more so than other organs? To resolve this question, our lab seeks to elucidate the molecular and cellular roles of tRNA modification enzymes in human health and disease. We have recently uncovered a novel tRNA synthetase-like mimic, DALRD3, that is required for a specific chemical modification in a subset of human tRNAs. Our preliminary results suggest that the DALRD3-dependent modification impacts tRNA conformational stability and function. Notably, we have also identified an autosomal-recessive variant in the DALRD3 gene that causes loss of function and the neurological disorder epileptic encephalopathy. Based upon these findings, we propose that DALRD3-mediated modification plays a critical role in the proper function of specific tRNAs important for protein synthesis during neurodevelopment. In our first Aim, we will define the requirements for tRNA recognition and modification dependent in DALRD3 and its cognate tRNA substrates. For our second Aim, we will measure the impact of DALRD3-dependent modification on tRNA structure and function. In our final Aim, we will determine the role of DALRD3-dependent tRNA modification on global protein translation in the brain through ribosome profiling. In total, the proposed research will have broad implications in understanding how tRNA modifications can impact proper neurodevelopment. Although DALRD3 was an unexpected player in tRNA modification, we now have a new target to explore potential therapeutics for individuals suffering from epileptic encephalopathies linked to tRNA biology.

NIH Research Projects · FY 2025 · 2022-09

The brain’s transport system for cerebrospinal and interstitial fluid, the glymphatic system, was first described in 2012 by the Nedergaard team, operates primarily during sleep, and has been linked to pathological neurological conditions including Alzheimer’s disease, traumatic brain injury (TBI), and stroke. Obtaining quantitative measurements of glymphatic fluid velocity and pressure is crucial to understanding the function, failures, and potential rehabilitation of the glymphatic system. However, existing techniques for obtaining in vivo glymphatic velocities are limited to sparse measurements and specific regions, and pressure variation is essentially impossible to measure in vivo. We propose to quantify glymphatic flows from measurements of tracked particles and contrast agents using physics-informed neural networks (PINNs), which can infer velocity and pressure from sparse measurements and have not been used previously in neuroscience. We will adapt PINNs for three commonly-employed glymphatic imaging modalities: two-photon perivascular space imaging, transcranial whole-brain imaging, and dynamic contrast-enhanced magnetic resonance imaging (DCEMRI). For these modalities, each of which can probe different regions and scales of glymphatic flows, we will adapt the PINNs equations and artificial intelligence hyperparameters, evaluate the sensitivity of the approach to noise, spatiotemporal resolution, and imaging artifacts using synthetic data, and validate by comparing velocities inferred by PINNs to velocities from alternative techniques. Using PINNs will allow us to obtain in vivo velocity and pressure measurements of cerebrospinal fluid in previously unmeasured regions of the brain. Our collaborative team of neuroscientists, fluid dynamicists, and applied mathematicians includes the leaders who discovered the glymphatic system and invented PINNs. Moreover, we have extensive experience with all three imaging modalities and with velocity measurement (via automated particle tracking and front tracking) in glymphatic flows. This proposal seeks to reveal mechanisms by which the brain's transport system for cerebrospinal and interstitial fluid operates. Our novel velocity and pressure measurements of intracranial cerebral spinal fluid flows may demonstrate how improving sleep, the state during which the glymphatic system primarily operates, can counteract pathological processes related to glymphatic system failure including Alzheimer's disease.

NIH Research Projects · FY 2025 · 2022-09

Hypertension is the single most important, medically modifiable risk factor for the prevention of cardiovascular disease in the United States. Control of hypertension is critical to improving the length and quality of life in the United States and for addressing racial disparities in cardiovascular disease. Yet, national progress in controlling hypertension has stalled. The current model for hypertension care in the United States, which relies nearly exclusively on clinician-driven office visits, has proven inadequate. There is an urgent need for team- based, patient-centered models of care. Team-based home blood pressure monitoring (TB-HBPM) represents an evidence-based practice that is widely underused in primary care. Strategies are needed to promote its adoption in primary care. Based on published barriers to adoption of TB-HBPM, successful strategies must engage patients and clinicians in the implementation process, and provide patients and their care teams with the knowledge, skills, resources, and data needed to implement and sustain TB-HBPM. Notably, strategies must address financial sustainability. The primary goal of this proposal is to identify and rigorously evaluate translatable strategies for implementing and sustaining TB-HBPM within primary care. To accomplish this aim, we will recruit seven practices from a single site where hypertension control is suboptimal. These practices serve predominately low-income and minority patients. In phase1 (R61), we will convene a steering committee that includes patients, practice staff, and clinicians to guide planning, implementation, sustainability, and evaluation (Aim 1). During phase 1, we will assess the specific barriers and facilitators to implementing TB-HPBM within these practices. Based on these practice-specific barriers, we will operationalize strategies using the Practical, Robust, Implementation, and Sustainability Model (PRISM). In phase 2 (R33), we will deploy these implementation strategies using a hybrid type-2, stepped wedge cluster randomized trial (Aim 2). Implementation strategies will include patient and team training, actionable data provided to the teams, and adoption of new billing codes. We will assess the impact of implementation strategies using the Reach, Effectiveness, Adoption, Maintenance (RE-AIM) framework (Aim 3). Our primary outcomes will be HTN control and patient use of HBPM. Secondary outcomes will include the proportion of patients with uncontrolled BP who are seen within 60 days, establishment of team charters by teams (adoption), and financial sustainability based on a cost analyses (maintenance). We will use realist evaluation to test theoretical assumptions underlying the implementation strategies (Aim 4). This mixed-methods approach will allow us to develop transferable lessons for other settings. Our findings will advance the science on implementation of successful HTN management models and provide a roadmap towards broader implementation of TB-HBPM in primary care.

NIH Research Projects · FY 2024 · 2022-09

(PLEASE KEEP IN WORD, DO NOT PDF) Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Adult hematopoietic stem cells (HSCs) are known to rely on signals from neighboring microenvironmental cells, such as the endothelial populations. While the role of blood vessels in promoting metastatic progression of solid tumors has been extensively studied, their function in the initiation and progression of early myeloid disorders, such as myelodysplastic syndromes (MDS), is less defined. MDS is characterized by a block in HSC differentiation and progressive bone marrow failure, especially in the elderly. With our ageing population, >10,000 new cases are diagnosed in the US every year. Since many patients with MDS develop aggressive therapy-resistant acute myeloid leukemias (AML), there is a significant need to understand mechanisms promoting MDS progression. Extensive work has shown increased angiogenesis in MDS patient bone marrow biopsies, possibly induced by angiogenic factors secreted by MDS cells. Higher disease burden often correlates with increasing microvascular density. Endothelial niche-driven signals are known to be critical for survival and chemoresistance of AML. These observations indicate a functional role for aberrant angiogenesis in promoting disease progression. However, it is not known when the endothelial niche remodeling initiates during MDS progression, what are the molecular changes that occur within the endothelial cells over time, and if the newly formed blood vessels provide support for disease progression. Based on the extensive remodeling of the endothelial niche in MDS, and the requirement for endothelial niche-driven signals in AML that can develop from MDS, we hypothesize that the altered vascular bone marrow niche provides support for MDS initiation as well as growth and progression of MDS. To test this, we will determine the spatiotemporal changes in the bone marrow endothelial niche and determine the functional role of the altered endothelial cells on MDS progression. In the long term, our work may help in designing novel therapeutic approaches aimed at inhibiting endothelial cell remodeling in MDS.

NIH Research Projects · FY 2025 · 2022-09

The coronary blood vasculature provides the heart with oxygen and nutrients, and removes metabolic waste. Organization of this contiguous network requires the maturation of vascular endothelial cells (EC) into arterial and venous fates based upon their location in the heart. While many of the guidance factors that control vascular patterning have been defined, it is not clear how spatial information controls cell behavior and identity. The epicardium is a single layer of mesothelial cells on the surface of the heart that harbors an important population of cardiovascular progenitors. We previously reported that epicardial epithelial-to-mesenchymal transition (EMT) is required for coronary EC maturation. New preliminary data reveals profound EC patterning and specification defects upon disruption of the epicardium, culminating with the inappropriate localization of angiogenic ECs in the sub-epicardium. To define the cellular and molecular mechanisms linking epicardial EMT to EC patterning and maturation we performed single cell (sc) RNA-sequencing of epicardium-derived cells and ECs isolated from the embryonic mouse heart at key developmental timepoints. This study defined epicardium-derived “shepherding” and “guidepost” cells that express unique angiogenic chemokine signatures. We provide in vitro and in vivo evidence that suggest a common mechanism controls EMT and the expression of genes that encode important guidance cues. We also find that EC localization and arteriovenous fate specification may be controlled by a common molecular mechanism. Based on previously published and preliminary data, we hypothesize that EMT controls the expression and localization of epicardium-derived chemokines that coordinate coronary EC patterning and AV fate specification in the fetal heart. The current study will interrogate this novel paradigm of epicardium-directed coronary EC patterning (localization and branching) and maturation (arteriovenous specification). Here, we will use genetically modified mice, time-lapse live embryo multi-photon imaging, scRNA-seq and spatial transcriptomics, and cell and molecular biology approaches to: 1) Define a common mechanism regulating EMT and the expression of genes that encode EC guidance cues; and 2) Interrogate the mechanisms coordinating epicardium-directed EC patterning and AV fate specification. We expect these studies will provide important insights into the mechanisms that control vascular patterning. This study may also advance our understanding of the developmental origins of coronary artery disease, and lead to therapeutic strategies that stimulate revascularization and repair of ischemic heart tissue.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Overall Research Plan The Human Lung Biomolecular Multi-Scale Atlas Program (HuBMAP-Lung) The Human Biomolecular Atlas Program (HuBMAP) is entering the production phase, with an emphasis on progressive, high sensitivity and specificity assays, prioritizing spatial biomolecular analysis. Our Tissue Mapping Center (TMC) proposal will focus on one organ system, comprised of two lungs and related respiratory tract (trachea to alveoli) that for simplicity we refer to as the “Lung”. Our TMC will generate, standardize, and validate extensive data from high content, high-throughput imaging and `omics technology to produce systematic human lung tissue maps at high resolution. We will take a long view of an Atlas, as a collection of 2D and 3D maps containing images, tabular data, facts about multiple locations at varying resolution, and indexes of named objects keyed to coordinates of a locational grid analogous to latitude and longitude, to cartographically present the whole range of salient features of the human Lung. To accomplish this task, our center brings together Investigators from five institutions, the University of Rochester (URMC), University of California at San Diego (UCSD), Pacific Northwest National Laboratory (PNNL), University of Washington (UW), and University of North Carolina at Chapel Hill (UNC), each with complementary lung-focused academic interests, knowledge, and inter- consortia network connections and with internationally recognized expertise in lung and state-of-the-art biomolecular technologies. Our Lung data provided to the HuBMAP Data Portal in Phase I has been focused to single nucleus RNA-sequencing, chromatin availability and beginning spatial transcriptomics. In addition to further increasing representation of human diversity in these data types, the next phase of our TMC turns primary effort toward determining spatial organization of cells and matrices defined by not only gene expression but also proteins, protein modifications, lipids and select metabolites critical to specific cell function within anatomical and functional niches. With the recognized value of global investigative efforts within and outside the Consortium, the Specific Aims of the TMC Overall Component concentrate on Communication and Collaboration within the TMC, between all HuBMAP components and synergistically among national and international researchers and Consortia.

NIH Research Projects · FY 2024 · 2022-09

1 Project Summary / Abstract 2 This career development project will provide a path for a highly qualified candidate with a combined MD/PhD 3 degree into a career as an independent investigator in reproductive science. Female infertility due to disorders 4 of androgen excess or insufficiency is a growing problem in the U.S. and worldwide with high mental and financial 5 costs. Androgen imbalance is implicated in polycystic ovary syndrome (PCOS) and diminished ovarian reserve 6 (DOR); however, while the phenotypic effects of androgen activity in the ovary are significant, little is known 7 about the underlying molecular pathways. This proposal aims to elucidate the mechanisms of androgen actions 8 in ovarian follicle growth and atresia, which are disordered in PCOS and DOR. The research direction stems 9 from the recent finding by the candidate that the androgen-regulated mRNA transcriptome in ovarian granulosa 10 cells is surprisingly small, and focuses on non-coding RNA regulation by androgens in granulosa cells. The 11 proposal also builds on prior studies of paxillin as a mediator of androgen signaling in prostate cancer. The role 12 of paxillin as a regulator of the androgen receptor expression is assessed for the first time in granulosa cells in 13 this proposal. The effects of paxillin loss in granulosa cells on follicular development and fertility will be examined 14 in a granulosa cell-specific knockout mouse model. The molecular mechanisms involving microRNA regulation 15 by androgens in granulosa cells will be studied in cultured mouse and human cells. The hypothesis is that 16 androgen effects in granulosa cells are mediated in large part by microRNAs, and are enhanced by paxillin. The 17 project has the potential to suggest novel therapeutic avenues for PCOS, a disease with sub-optimal 18 management options. The training plan capitalizes on the applicant’s strong research background and long- 19 standing interest in female fertility. She aims to build on her technical skills, develop as a leader and a member 20 of the scientific community, and transition to independence in research. The mentor is a physician-scientist who 21 is a world expert in paxillin and androgen signaling, mouse genetics and ovarian biology with a long track record 22 of mentoring successful trainees. The training will take place in a highly collaborative environment where 23 expertise in reproductive research techniques is abundant. The applicant has assembled an advisory committee 24 comprised of world experts in ovarian biology, paxillin, steroid signaling, and microRNA sequencing. She will 25 take advantage of University-sponsored courses in translational research methods, leadership and professional 26 development, as well as research seminars, national meetings and one-on-one instruction. At the end of this 27 project, she plans to write several first-author publications, run a lab independently, mentor a graduate student 28 and apply for an R-01 grant.

NIH Research Projects · FY 2024 · 2022-09

Timbre, the quality that allows sounds to be distinguished when they are identical in pitch, level, and duration, is a critical aspect of speech comprehension and music enjoyment. My proposal will fill a gap in neural studies of timbre by testing the hypothesis that capture and off-CF inhibitory mechanisms lead to robust representations of suprathreshold synthetic and natural-instrument timbre in the midbrain. To test my hypothesis, I will record single-unit inferior colliculus (IC) responses from awake Dutch-belted rabbits. I will also develop a new computational IC model based on these physiological responses. Spectral envelopes of harmonic sounds are correlated with the timbral perception of “brightness”. I propose two mechanisms that contribute to spectral-envelope encoding: capture and off-characteristic frequency (CF) inhibition. The first mechanism, capture, refers to the dominance of harmonics near spectral peaks over auditory-nerve fibers tuned near the peaks. Capture is due to saturation of inner hair cells. Capture by a single harmonic reduces the amplitude of low-frequency neural fluctuations in auditory-nerve fibers. Rates of IC neurons are sensitive to low-frequency neural fluctuations as characterized by modulation transfer functions. Ultimately, capture of auditory-nerve responses for fibers tuned near spectral peaks results in IC rate profiles that encode spectral peaks. Preliminary results are partially consistent with spectral peaks of synthetic timbre stimuli capturing peripheral responses, leading to a rate representation of salient spectral features in the midbrain. However, another mechanism that could explain IC representations of timbre is off-CF inhibition, which has been proposed to explain frequency-sweep sensitivity and psychophysical forward masking. A subcortical computational model that features capture, but not off-CF inhibition, was able to predict preliminary responses to synthetic timbre and narrowband tone-in-noise, but could not predict responses to wideband tone-in-noise or natural timbre, indicating the need to update the model. I have developed experiments to test the hypothesis that timbre is robustly encoded in the midbrain via capture and off-CF inhibition. Aim 1.1 will test the hypothesis that responses to wideband tone-in-noise are strongly influenced by off-CF inhibition, and reducing the noise bandwidth increases the influence of capture. In Aim 1.2 I will update a computational IC model by adding off-CF inhibition to test the hypothesis that capture and off-CF inhibition are necessary to explain tone-in-noise stimuli. Aim 2.1 will test the hypothesis that the spectral peak of a shaped harmonic complex, synthetic timbre, is robustly encoded in the IC over a range of suprathreshold sound levels. Aim 2.2 bridges the gap between synthetic and natural timbre by recording responses to real instrument sounds. Responses from Aim 2 will further test the new IC model. This project will provide insight on the IC representation of suprathreshold timbre. This research will lead to novel strategies to restore timbre perception in hearing aids and cochlear implants, which are not designed for timbre perception.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY This Mentored Clinical Scientist Research Career Development application will support Dr. Matthew McGraw in his transition to independence as a clinician scientist studying the mechanisms of airway basal cell dysfunction in chemical-induced bronchiolitis obliterans (BO). BO is a devastating fibrotic airways disease, most commonly seen after organ transplant. However, BO is becoming more frequently associated with inhalation exposures to certain viruses or chemicals. One of the most well-known chemicals associated with inhalation-induced BO is diacetyl (DA; 2,3-butanedione), a highly reactive diketone found in foods, coffee and e-cigarettes. Despite DA’s common use as a flavoring additive, the mechanisms of DA-induced BO remain poorly understood. Central to BO development is injury to the airway epithelium. When injured, the airway relies on epithelial progenitor cells for proper repair. The objective of this application is to better understand the functional role of airway basal cells, the primary progenitor cell of the human airway, in chemical-induced BO. Two preclinical models of chemical- induced BO were developed for this application. First, rats exposed consecutively to DA vapors developed persistent hypoxemia, reduced weight gain, and histologic evidence of BO. Poly-ubiquitinated proteins accumulated in rat airways after DA exposures not seen in air controls. Second, in human airway epithelial cells exposed to DA vapors, poly-ubiquitinated proteins accumulated and co-localized primarily with airway basal cells. With repetitive DA exposures, the accumulation of polyubiquitinated proteins resulted in proteotoxicity of airway basal cells. Our central hypothesis is repetitive DA vapor exposures results in abundant protein damage, leading to proteotoxicity of airway basal cells, impairing airway epithelial repair and promoting BO development. Aim I of this proposal will determine how abundant protein damage in airway basal cells impairs epithelial repair and promotes BO development using both models of repetitive DA vapor exposure. Aim II will determine the role of the ubiquitin proteasome system in airway basal cell toxicity and BO development. Aim III will compare the efficacy of multiple ubiquitin proteasome pathway drug targets in preventing basal cell toxicity and BO development. Dr. McGraw has assembled a mentoring team of experts in the fields of airway epithelial biology (T Mariani, PhD; primary), inhalation toxicology (JN Finkelstein, PhD; I Rahman, PhD), and proteomics (WJ Qian, PhD) for critically examining the role of airway basal cells in chemical-induced BO. Mentoring in airway stem cell biology and proteomics, as described in this proposal, will facilitate Dr. McGraw’s transition to independence. At K08 completion, the data generated from this application will significantly advance our understanding of airway basal cell function following inhalation exposures and have a broader impact on neighboring research fields of inhalation toxicology and BO development.

NIH Research Projects · FY 2025 · 2022-09

Abstract: RNA sequences have pervasive roles in biology and RNA molecules are also important pharmaceuticals. Many of these RNA molecules function by use of specific structures, and these structures are therefore conserved across evolution. With the availability of high throughput methods to identify RNA transcripts and probe structure, there is a need to model these structures to get the most information from available data. The goal of the Mathews lab is to develop algorithms and software to model RNA structures, including secondary and tertiary structures. We focus on using biophysical principles, and we are also at the forefront of incorporating additional information in our models, including experimental mapping data and structure conservation. Here, we will build on the foundation of prior work to develop new structure prediction methods. We provide our software freely to the community and we rigorously test our methods with close collaborations with experimentalists.

NIH Research Projects · FY 2024 · 2022-08

Project Summary and Abstract Fetal weight estimation, or the assessment of antenatal fetal weight for the purposes of growth tracking and labor planning, is a critical component of safe prenatal care. Estimations currently rely on ultrasound-derived measurements of specific fetal planes to indirectly assess growth and wellbeing. The standard fetal biometric measurements for the estimation of fetal weight (biparietal diameter, head circumference, abdominal circumference and femur length) are poorly correlated to actual fetal weight, defined as the measurement of newborn weight in grams at birth. For newborns who are above 4,000 grams at birth, current error estimates of fetal weight in the late-third trimester of pregnancy are only accurate approximately 40% of the time. By no longer relying on fetal biometric measurements, data science approaches have the potential to estimate fetal weight with lower bias and errors compared to standard regression methods. To date, no studies have used ultrasound images, not just the fetal measurements, as input into a neural network approach to estimate fetal weight. The overarching goal of this proposal is to develop the skills and training necessary to lead the advancement of data science for use in clinical assessment during pregnancy. Using existing ultrasound imaging and birth certificate data (n=17,478 patients) from the University of Rochester (UR) Medicine Hospitals and the Finger Lakes Regional Perinatal/Obstetrics Data System (PDS), and n= 310 patients in the R01 study, Understanding Pregnancy Signals and Infant Development (UPSIDE: R01HD083369), the specific aims are: 1) To determine the maternal (i.e., body mass index) and fetal factors (i.e., growth measurements) that increase the discordance between the estimation of fetal weight by the Hadlock formula and actual birth weight of neonates using birth certificate data from the PDS, 2) To evaluate the accuracy of a CNN algorithm on ultrasound images in the third trimester to estimate fetal weight compared to the Hadlock formula, and 3) To test the effectiveness CNN algorithm on new ultrasound images from the UPSIDE study. This proposal will leverage the expertise of Dr. Caitlin Dreisbach’s mentorship team, computational resources, and the exceptional research environment at the UR School of Nursing, Goergen Institute for Data Science, and the Rochester Institute of Technology. Results from this study have the potential to change practice and improve clinical assessments during the late third trimester of pregnancy. The research study and mentored training included in this award allows Dr. Dreisbach to establish her long-term career goal of becoming an independent investigator with expertise in the translation of data science to obstetric clinical care.

NIH Research Projects · FY 2025 · 2022-08

Resolving amygdala microcircuits: implications for function Years of neuroimaging research in human psychiatric disorders, including at-risk populations, highlights alterations in circuits involving the amygdala. 'Fingerprints' of specific cortical and amygdala correlated activity promise to help identify specific circuits involved in particular diseases. Despite this, the structural or 'wiring' details of cortical-amygdala circuits remain obscure. Here we examine the idea that the unique symptom profiles across heterogeneous disorders are best explained by how different combinations of cortical inputs form 'microcircuits' in the amygdala, each associated with unique output paths. This proposal focuses on cortical nodes that project to the amygdala, and are frequently involved in psychiatric conditions. The subgenual anterior cingulate (sgACC) and rostral agranular insula (Ia) mediate internal body sensations, or 'interoceptive states'; the perigenual ACC (pgACC) and the middle, dysgranular insula (Id) are regions involved in 'social monitoring' and affiliative responses. Using dual anterograde tract tracing approaches in the same animal, our preliminary data show that afferent terminals from pgACC 'social monitoring' nodes are always 'nested' in terminals arriving from the sgACC ('interoceptive) in the amygdala. sgACC terminals cover a broader territory. This overlap creates terminal 'hotspots' of sgACC and pgACC afferent integration. At the cellular level, sgACC and pgACC associated terminals almost always co-contacted pyramidal cells (rather than segregating to different populations), and did so in a very stable ratio. These data indicate normal, tight afferent control, and integration of incoming information, onto amygdala neurons in these 'hotspots'. This terminal balance suggests an anatomic substrate that may be altered in various disease states. We also found that amygdala neurons in converging sgACC/pgACC hotspots had distinct projections back to the cortex, which were different than from non-hotspots regions. In this proposal, we dissect microcircuits involving sgACC, Ia, pgACC, and Id. We begin with determining specific cortical networks involving these nodes (Aim 1), then examine how afferent terminals from different two-node systems overlap or segregate in the amygdala, including at the cellular level (Aim 2A, B). We then examine whether amygdala neurons in converging afferent 'hotspots' have unique outputs to the cortex (Aim 3).

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT: Key components of skeletal muscle that regulate excitability and excitation-contraction coupling (ECC) undergo major shifts of isoform expression during development. This process of perinatal ECC remodeling is highly conserved throughout vertebrate evolution and results mainly from post-transcriptional mechanisms in which alternative splicing of specific exons for ClC-1, CaV1.1, RyR1 and SERCA1 occurs. In myotonic dystrophy (DM), these splicing switches revert to their fetal set points due to sequestration of MBNL splicing factors in nuclear RNA foci. We used gene editing to recreate individual DM splicing defects in mice and systematically analyzed mice for effects in isolation and in combination through breeding. Our preliminary studies indicate that loss of ClC-1 function combined with CaV1.1 exon 29 exclusion (Cav1.1∆e29), comparable to that observed in DM patients, results in severe muscle weakness and respiratory deficits and is lethal in mice by age ~9 wks. This effect is rescued by long-term treatment by oral feeding with a Food & Drug Administration (FDA) approved calcium channel blocker. Here we propose studies to define mechanisms and explore the possibility that drug treatments that target myotonia and Cav1.1 channels can mitigate muscle weakness in DM. In Aim 1 we will investigate the mechanism for the early demise of myotonic Cav1.1∆e29 mice, including the study of how Cav1.1∆e29 channels impact membrane excitability and Ca2+ homeostasis, and downstream effectors that include calpain, transcription and mitochondrial health. Further, we will determine if myotonic Cav1.1∆e29 mice exhibit skeletal muscle weakness and altered respiratory function. In Aim 2 we will treat myotonic Cav1.1∆e29 mice by oral feeding of FDA approved drugs that target the calcium channel or myotonia by factorial design (one, the other, both or neither) to see which treatment is most effective at extending life and improving muscle and respiratory function. In Aim 3 we will move the treatment into a CUG repeat expansion DM1 mouse model that exhibits severe muscle weakness, myopathic features and shortened lifespan. We will use a series of non- invasive techniques to measure muscle and respiratory function to determine treatment benefit in longitudinal studies. The ultimate goal of this proposal is to identify DM1 therapeutic interventions that can be rapidly transitioned to the clinic.

NIH Research Projects · FY 2025 · 2022-08

Project Summary/Abstract As many as 10 million adults in the United States have temporomandibular disorders (TMDs). Females experience TMDs at higher rates compared to males and African Americans have higher TMD incidence, yet lower prevalence compared to white Americans. Despite these findings, etiological mechanisms contributing to prevalence and incidence disparities by sex and race are largely unknown. Given our recent reports on sexual dimorphisms in incremental tensile stiffness and fixed charge density of the human temporomandibular disc, it is hypothesized that additional sexual dimorphisms may exist in properties of the temporomandibular joint (TMJ). Similar differences are also hypothesized to exist between racial groups, although this possibility has not yet been explored, which may limit research translational confidence across racial groups. The lateral capsule- ligament (LCL) complex, which spans the temporal bone and mandibular condyle, could impact temporomandibular disc derangement (TMDD) risk through a ‘loose ligament’ mechanism affecting joint articulation postulated in prior literature. Furthermore, sex- and race-specific differences in potential risk and resiliency factors may contribute to differential risk of TMDD development and observed TMD prevalence and incidence disparities. Our long-term goal is to enable individualized TMDD risk assessment to increase applicability of generated risk estimates. The objective of this study is to describe sex- and race-specific mechano-chemical LCL complex properties to investigate potential contributions to TMD disparities between sexes and racial groups. The central hypothesis is that TMJ properties observed in females and African Americans will be associated with higher risk of TMDD development compared to those of males and white Americans. The proposed studies will enable the determination of sex- and race-specific differences in potential mechanical and chemical risk factors for TMD development (Aims 1 and 2). This determination will contribute to a better understanding of TMJ properties across diverse populations and is crucial to enabling understanding of TMD incidence and prevalence disparities. Finite element models will also be used to investigate how differences between sexes and racial groups impact the temporomandibular loading environment, which will be developed, refined, and interpreted during the independent phase (Aim 3). This proposal aims to further development of scientific expertise and skill acquisition in temporomandibular and craniofacial biomechanics, with a plan to transition from mentored post-doctoral fellowship to independent faculty. NIH T32 training and professional development resources, joint resources and network of the Clemson- MUSC Bioengineering program, and the Clemson University Pathways Mentoring Program provide an ideal inter-disciplinary training environment to prepare for and transition to independence following the postdoctoral fellowship period.

NIH Research Projects · FY 2025 · 2022-08

Clinical and epidemiological studies have consistently connected HIV infection with increased risk of cardio- vascular disease (CVD). This is due to dysregulated health status associated with residual virus replication, release of soluble viral proteins, and HIV-induced immune activation, which may also be associated with premature immune senescence and persistent inflammation. Chronic inflammation is associated with acceleration of atherosclerosis (AS) and is increasingly prominent among the HIV+ individuals on cART. Monocytes respond to inflammatory stimuli and are precursors of the macrophages within atherosclerotic lesions, including lipid-laden foam cells, and lesion-associated macrophages represent a major source of a number of cytokines and chemokines that direct monocytes into vascular lesions, thus creating a positive- feedback loop. Differentiation of monocytes is enhanced upon interaction with platelets. We and others have shown that platelet-monocyte complexes (PMCs) are increased during HIV infection and CVD. Therefore, we propose to investigate a hypothesis that the PMCs promote atherogenic processes in persons with HIV by triggering monocyte differentiation through horizontal transfer of micro-RNA and protein encapsulated in the platelet-derived microparticles (PMs) to the monocytes and thereafter their interaction with endothelial cells. To test these hypotheses in first aim we define and model molecular networks in monocytes that have or have not interacted with activated platelets in HIV+/- patients with/without carotid plaques using state-of-art CITE-seq. We will employ discrete-state modeling approaches to identify pathways and subnetworks dysregulated in HIV+ individuals with carotid plaques. In second aim, we propose to characterize micro-RNA (miRNA) and proteins encapsulated in platelet-derived microparticles by small-RNA sequencing and proteomics in HIV+/- individuals with/without carotid plaques. The list of miRNAs and proteins will be integrated with the network model from aim 1 to simulate the effect on signaling cascades and their cross-talk. The network model will reveal the role of miRNAs and proteins on the monocyte differentiation into pro-inflammatory phenotype. Additional analysis by machine-learning techniques will allow further prioritization and selection of genes. The ex-vivo experiments will test binding to human carotid endothelial cells and receptiveness of endothelial cells to the differentiated monocytes. In conclusion, our multidisciplinary approach of integrated omics, computational modeling and ex- vivo experiments will reveal mechanisms by which AS is prematurely induced in HIV+ individuals, novel targets to curb AS in HIV+ individuals and biomarkers for early detection of AS. This study holds strong prospects to significantly improve aging related comorbidities in persons living with HIV in general and AS in particular.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY / ABSTRACT This R35 program aims to delineate a previously unappreciated role of Piezo1 mechanosensitive ion channels in regulating cellular activity under inflammatory conditions using articular chondrocytes (ACs) as a model system. Mechanotransduction is a highly dynamic and ubiquitous process, and virtually all mammalian cells exhibit rapid and robust ion influx upon mechanical stimulus. In disease states, cells present abnormal mechanotransduction revealed by altered mechanosensitivity and biochemical transduction. Piezo1 is a novel mechanically-activated (MA) Ca2+-permeating channel abundantly expressed in cardiovascular, musculo- skeletal, and neural tissues. Augmented functional expression of Piezo1 in ACs is associated with cartilage disorders. Interestingly, ACs present a morphological differentiation from spherical ACs to cells with point-like processes in osteoarthritis. The actin filament (F-actin) networks have been focused on understanding morpho- logical differentiation and mechanical stability of AC, yet our preliminary data suggest a potential role of Piezo1 in F-actin remodeling in process formation under IL1α-mediated inflammatory conditions. Here, we propose a paradigm-shifting concept of Piezo1-mediated morphological changes and pro- cess formation in inflammatory conditions. Both mechanotransduction and inflammation influence or are re- sponsible for a wide array of physiological processes and abnormalities. A knowledge gap exists in the mecha- notransduction-inflammation coupling, morphology-dependent cellular mechanosensitivity, Piezo1-mediated morphological changes, Piezo1 localization on the membrane cortex of round-shape ACs, and Piezo1-recruit- ment in processes of fibroblast-like shaped ACs. This proposed program will identify: (i) if activated Piezo1 al- ters local membrane curvature and triggers to form cellular protrusions and reorganize cytoskeleton meshwork, (ii) specific inflammatory cytokines triggering morphological changes and augmenting Piezo1 channels in pro- cesses, which in turn altering the gating properties of Piezo1, and (iii) if Piezo1-mediated inflammatory re- sponse increases mechanical fatigue and death. The whole-cell patch-clamp method allows to record ion channel activities, yet ACs present technical challenges on sealing the membrane primarily due to their round morphology and small size (~10 μm in diameter). Thus, we have custom-built a novel microscopy, named ‘Mechano-microscopy’, by combining an atomic force microscope (AFM), a high-speed ratiometric Ca2+ im- aging microscope, and a confocal microscope to simultaneously record cellular morphology and biochemical signals in response to mechanical cues under inflammatory conditions. In addition, we will utilize the super- resolution STED microscope and Piezo1-TdTomato reporter mice to interrogate dynamics of membrane curva- ture and Piezo1 localization in ACs after treating specific inflammatory cytokines. Our program will conceptually unveil fundamental processes of Piezo1-dependent inflammatory re- sponses leading to cellular morphological differentiation. In addition, our results will provide new insights into the mechanotransduction-inflammation coupling mechanisms and lead to new therapeutic strategies to prevent morphology changes at the early stages of cartilage disorders.

NIH Research Projects · FY 2025 · 2022-08

Program abstract: This proposal aims to identify the neural circuit mechanisms that control periarterial cerebrospinal fluid (CSF) pumping and glymphatic clearance of fluid and solutes. We have developed a collaboration to quantify CSF transport dynamics in both humans and mice across several scales, spanning molecular transport, neuronal and glial activity, vascular and brain-wide fluid dynamics. We propose that coordinated neural activity during sleep drives global and local changes in blood volume, which in turn are the primary drivers of CSF transport. Our model establishes a novel conceptual framework, namely that neuronal circuits control clearance via their effects on astrocytes and the vasculature, opening an array of testable hypotheses across spatial scales and species. Project 1 will build quantitative fluid-dynamical models to establish how arterial dilation, mediated by neural activity, drives periarterial CSF pumping and glymphatic efflux across length scales. Models for both mice and humans, informed by experiments in Projects 2-4, will drive hypotheses to be tested in those Projects. Project 2 will dissect how neural activity transmits Ca2+/cAMP signaling to the neurovascular unit, thereby altering the physical dimensions and functional properties of the perivascular spaces. Viral tagging combined with optogenetic stimulation of individual cell populations will reveal neural effects on CSF flow, measured by particle tracking. The Project will also provide the first systematic analysis linking periarterial CSF inflow with glymphatic solute clearance. Project 3 will dissect the local neural and global neuromodulatory drivers of vasodynamics during NREM sleep using optogenetic and chemogenetic manipulations. Additionally, local and global arterial dynamics during sleep will be imaged, providing key information on the vascular pumping of CSF movement. Project 4 will use novel MRI-based techniques to establish how neural activity and large-scale fluid flow are linked in the human brain. By driving local neural activity with sensory stimulation, and imaging spontaneous neurovascular and CSF dynamics across arousal states, it will test how specific spatiotemporal patterns of neural activity affect hemodynamics and CSF flow in wakefulness and NREM sleep. The Projects will be supported by Cores focused on Viral Tools, Data Science, and Administration, all overseen by Internal and External Advisory committees. Together, the Projects will provide a quantitative, circuit-based understanding of the neural mechanisms governing brain fluid flow and solute clearance during sleep.

NIH Research Projects · FY 2025 · 2022-08

1. PROJECT SUMMARY/ABSTRACT This NIA career development award will establish Dr. McGarry, an Assistant Professor in the Department of Medicine at the University of Rochester (UR), as an independent investigator with expertise in the areas of cognitive aging, including Alzheimer’s Disease and related dementias (ADRD), and consumer choice in the Medicare program. With support from the award, Dr. McGarry will undertake a training plan that focuses on: 1) the clinical trajectory of ADRD, 2) ADRD family caregiving and research, 3) qualitative research methods, and 4) leadership of a multidisciplinary research team. This training will enable Dr. McGarry to conduct multidisciplinary, mixed-methods research examining the impact of ADRD on Medicare choices and resultant outcomes. It builds off his current areas of expertise, including Medicare policy and benefit design and quantitative health services research. Dr. McGarry has leveraged the considerable resources available at UR to assemble a multidisciplinary team of nationally-recognized experts. Dr. Kathi Heffner, the primary career development mentor, is a social psychologist with expertise in cognitive aging and the study of caregivers with ADRD. Dr. Helena Temkin-Greener, the primary methods mentor, is a health services researcher with expertise in the delivery and quality of health services for older adults and mixed methods research. Together they will lead a mentorship team with additional expertise in qualitative research methods, the measurement of functional cognitive abilities, the diagnosis and clinical care of individuals with ADRD, and the use of national survey data linked with Medicare claims to study health outcomes in older adults. The Medicare program depends on consumer choice, yet little is known about the ability of older adults with ADRD to navigate this complexity. Available evidence suggests this population is at increased risk of suboptimal coverage choices, yet little is known about how these risks evolve over the disease course and their health and financial effects. This mixed-methods study will examine the Medicare coverage choices of individuals with ADRD throughout the disease course (Aim 1). The Health and Retirement Study (HRS) linked with Medicare claims will be used to describe the Medicare coverage, and propensity to make changes, of individuals with ADRD at various stages of cognitive decline. Analyses will also examine whether the impacts of cognitive declines are moderated by the availability of family caregivers and socioeconomic resources. Qualitative interviews with caregivers of individuals with ADRD will add depth and support the interpretation of the quantitative results. Using the same HRS data, the effects of impaired household Medicare decision making due to ADRD on health and financial outcomes will be estimated using a quasi-experimental design (Aim 2). Study results will provide critical information to policy makers about the effects of Medicare program design on the well-being of individuals with ADRD. In combination with mentored training, this research will form the basis of an R01 proposal that will evaluate the effects of potential policy reforms designed to support Medicare choices in households with ADRD and the acceptability of such reforms to this population.

NIH Research Projects · FY 2026 · 2022-07

Project Abstract Mutated clones in hematopoietic cells, also known as clonal hematopoiesis (CH) are present in healthy individuals and expand with aging. In spite of normal hematopoietic parameters, individuals with CH have increased risk of myeloid neoplasms, cardiovascular risk and all-cause mortality. We recently showed that the aged microenvironment contributes to hematopoietic stem cell fate choices. While the presence of clonal hematopoiesis in healthy individuals has been widely reported, and its expansion with aging is established, whether the aging microenvironment modifies clonal dynamics and contributes to clonal selection leading to progression to myelodysplastic syndromes (MDS) remains unexplored. Sirtuin6/SIRT6 is a regulator of genome and epigenome stability. SIRT6 is responsible for more efficient DNA repair in long-lived species. Moreover SIRT6 plays a critical role in suppressing retrotransposon expression, demonstrating that retrotransposon activity directly contributes to the progeroid phenotype in mice lacking SIRT6, in part through activation of innate immunity. Nucleotide reverse transcriptase inhibitors (NRTIs) developed as HIV drugs, inhibit retrotransposition, reduce inflammation, improve aging biomarkers in wild type mice, and extend the lifespan of progeroid mice lacking SIRT6. We hypothesize that aging-associated de-repression of retrotransposons promotes pro- inflammatory changes of specific hematopoietic stem cell-supportive niche populations (marrow macrophages and multipotent stromal cells) which drive clonal progression to myeloid neoplasms. To test this hypothesis, using relevant models of clonal hematopoiesis we will examine whether 1) pre-leukemic mutations form clones and progress to MDS more readily in the aged microenvironment by transplanting them into young versus aged recipient mice; 2) SIRT6 overexpression in key microenvironmental populations slows the rate of microenvironmental and hematopoietic aging, clonal expansion and progression to MDS; 3) repressing LINE1 retrotranspositions with inhibitors of reverse transcriptases (NRTIs) slows clonal expansion and provides a mechanism to discover novel microenvironmental regulators of clonal hematopoiesis progression.

NIH Research Projects · FY 2026 · 2022-07

PROJECT SUMMARY Developmental progression from cleavage phase to gastrulation occurs during the mid-blastula transition (MBT). This transition coincides with several critical events, including maternal to zygotic transition, zygotic genome activation (ZGA), and stem cell formation. How the zygotic genome becomes activate and how proper developmental timing is regulated are critical unknowns in biology. Recent data in zebrafish and Drosophila indicate that these processes are highly sensitive to the levels a particular histone, the histone variant H2A.Z (H2Av in Drosophila). However, it remains largely unknown why altered H2A.Z levels disrupt MBT events and how the embryo ordinarily assures that the correct amount of H2A.Z is incorporated into chromatin. Preliminary observations indicate that in both species a subset of zygotic genes is prematurely activated when H2A.Z levels are elevated. A combination of genomics approaches and expression analysis of candidate genes will be used to determine the temporal pattern of gene activation in embryos with up- or downregulated nuclear H2A.Z levels. It will also be tested to what extent the observed changes are due to chromatin associated factors (in particular the pattern of H2A.Z distribution across the genome) and epigenetic marks. Use of the two distinct model systems will reveal to what extent the underlying mechanisms are conserved between vertebrates and insects. In Drosophila, it is known that H2A.Z levels in the nucleus can be regulated by the H2A.Z binding protein Jabba that sequesters H2A.Z in the cytoplasm, a process active during the time of the MBT. Preliminary studies have led to the hypothesis that a different H2A.Z-binding protein plays an analogous role in zebrafish. This hypothesis will be tested using live imaging, embryo injections, and structure-function analysis. Successful completion of this project will define the role of H2A.Z in controlling gene expression patterns and in cellular programming during one of the most crucial periods of development, the mid- blastula transition.