Case Western Reserve University

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY/ABSTRACT Northeast Ohio has one of the highest rates of oral diseases (caries, periodontitis) and poor dental attendance among low-income older adults. Professional organizations and the IOM recommend integration of oral health (OH) activities into primary care for adults in order to reduce medical costs. But, published literature indicates a lack of outcomes data to assemble an effective medical-dental integration. Impeding integration are also factors such as lack of an electronic health record (EHR) based oral health (OH) assessment and referral, and inadequate OH education and training for medical providers. Our survey data indicate that the majority of providers would like OH facts to be communicated at primary care visits (PCV) but lack education and resources. There are misperceptions about oral diseases among older adults that prevent regular dental attendance. The proposed multi-level interventions will address factors that impede OH integration, and subsequently improve self-regulatory behaviors in adults. The interventions are: Practice (medical assistants, nurses): EHR systems based changes to ask, advise, assess, connect (AAAC). Provider (physician/nurse practitioner): improve knowledge and skills using Common-Sense Model of Self-Regulation (CSM) theory based education and skills training to communicate OH facts and reinforce importance of dental visits. A cluster-randomized clinical trial is proposed to test implementation (practice) and behavioral (provider) intervention to address self-regulation and increase dental attendance among low-income adults aged ≥55 years. The primary aims are: 1) UG3, Conduct qualitative work with stakeholders and practices; system-based changes in EHR; and pilot-test the interventions in 2 practices. UH3, randomize 8 practices to two arms to investigate the efficacy of a EHR based strategy at the practice level to ask [OH risk assessment], advise [going to dentist], assess [willingness for referral], connect [eReferral and/or resources] together with provider CSM theory-based education and skills to communicate OH facts versus provider alone (standard or usual oral health care) to increase dental attendance (primary outcome); and improve OH quality of life, oral hygiene behavior, and biometric measures of health (secondary outcomes). Secondary aims (UH3) are to explore: the delivery and documentation of AAAC implementation strategy; and to investigate causal pathways that affect the outcomes. The sample includes 209 providers and medical staff, and 800 Medicaid-enrolled adults. Data analysis (UG3) will utilize a mixed method design for qualitative and descriptive statistics for quantitative data. Data collection (UH3) will follow the RE-AIM framework: adults (outcome data from Medicaid claims, questionnaires, EHR); provider, practice (questionnaires); provider, practice (process measures: reach, fidelity, adoption, maintenance from audits). A generalized estimating equations approach will be used to assess effects of multi-level interventions on dental attendance and other outcomes, while accounting for clustering within practice. Mediation methods will determine if intervention effects occur through hypothesized mediators. A sustainable OH care model is proposed for primary care clinicians.

NIH Research Projects · FY 2025 · 2022-06

ABSTRACT Systemic inflammation and mortality in people living with HIV (PLWH) are associated with mucosal immune dysfunction and persistent viral reservoirs in the mucosa and lymphoid tissues. In the previous project period, we have demonstrated the role of CD4+ FOXP3+regulatory T cells (Treg) dysregulation in contributing to aging- and HIV-associated oral inflammation. The dysfunctional Tregs that we uncovered in the oral mucosa of PLWH mimic tissue-resident CD4+ T cells expressing high levels of PD-1 and amphiregulin (AREG). These cells correlate with CD38+HLADR+ CD4+T cell hyperactivation and dysfunction, toll-like receptor 2 (TLR-2), and inflammasome/IL-1β signaling in vivo, and require IL-1β/AKT pathways for their expansion in the context of HIV infection. Here we propose to address the role of oral fungal dysbiosis in establishing the inflammatory environment and triggering oral immune cells to become dysfunctional. Building on our more recent results from salivary metabolome analysis in PLWH, we will examine the underlying pathways at the interface of dectin-1, AREG, inflammasome, and immunometabolism signaling pathways in three well-connected aims. Thus, our proposal will reveal new directions to manage oral immune-dysfunction and thereby systemic inflammation, and alter current clinical practice.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Alzheimer's disease (AD) is the most common form of dementia, affecting ~10% of the population over 65. Processing amyloid-beta precursor protein (APP) into amyloid-beta peptide (Aβ) is an important and fundamental aspect of AD pathogenesis. However, the signaling pathways that control this process are not well defined. Mitochondrial dysfunction is a prominent early feature in susceptible neurons in the brain of patients with AD and plays a critical role in AD pathogenesis. APP and its metabolic fragments are known to localize to mitochondria where they negatively influence mitochondrial function. Conversely, neurons with impaired mitochondrial function and bioenergetics also contribute to APP processing and synaptic loss. Although evidence suggests that APP-mitochondrial interactions are important for the cognitive deficits and amyloid genesis in AD, the field lacks a detailed understanding of the mechanisms that coordinate APP processing and functional mitochondrial deficiency. The coiled-coil-helix-coiled-coil-helix domain-containing protein 6 (CHCHD6) is an evolutionarily conserved nucleus-encoded mitochondrial protein. CHCHD6 is a core component of the mitochondrial contact site and cristae organization system (MICOS), which controls mitochondrial respiration and redox regulation, lipid homeostasis, and membrane ultrastructure and dynamics. Recent proteomics studies have shown a decrease in CHCHD6 in brain samples of AD patients and AD mice. However, the role of CHCHD6 in AD pathology is unknown. Our preliminary studies have revealed a previously unidentified role of CHCHD6 in the regulation of APP-mediated neuropathology and cognitive deficiency. The objective of this application is to determine the role of CHCHD6-mediated AD pathology at both mechanistic and therapeutic levels. In Aim 1, we will determine the causes and consequences of CHCHD6 loss of function in AD models. In Aim 2, we will dissect the mechanism of CHCHD6 loss in AD pathology. In Aim 3, we will determine whether compensation for the loss of CHCHD6 in AD mice alleviates neuropathology and cognitive deficits. If successful, these studies will establish CHCHD6 as a key molecule linking APP processing, lipid disturbance, and mitochondrial dysfunction and advance our understanding of AD pathology. The findings generated by this project will also have a significant impact on AD research by identifying CHCHD6 as a novel target for limiting mitochondrial dysfunction, amyloid pathology, and cognitive deficits in AD.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY/ABSTRACT Cardiomyopathy (CM) encompasses a diverse group of diseases of the heart muscle that occur in 1 out of 500 adults and predispose to heart failure. Accurate determination of CM subtype (phenotyping) is essential to inform downstream tests, risk stratification, and targeted treatments. Cardiac MRI has emerged as the non-invasive standard for assessment of cardiac structure, function, and tissue properties in patients with suspected CM. However, cardiac MRI only comprises 1% of all MRI exams in the US, largely due to the need for (1) long and complex protocols where multiple images are collected with different contrast weightings, (2) unreliable and uncomfortable strategies to reduce motion, and (3) a lack of reproducibility of certain tissue property measurements. This multidisciplinary project between MRI scientists and cardiologists will validate 3D cine Magnetic Resonance Fingerprinting (MRF) as a comprehensive all-in-one imaging technique for CM detection and phenotyping. A streamlined and paradigm-changing cardiac MRI exam is proposed, consisting of a 5-minute free-breathing and ungated 3D cine MRF scan that will be collected before and after administration of gadolinium contrast. This technique will yield quantitative T1, T2, and spin density (M0) maps with 3D isotropic coverage over the left (LV) and right (RV) ventricles. Additionally, measured tissue properties and MRI simulations will be used to generate contrast-weighted cine and LGE images in an automated fashion, eliminating the need for multiple acquisitions and manual scan adjustments. A multicontrast LGE approach is also proposed where bright-blood, dark-blood, and novel “optimal-contrast” images will be generated to optimally highlight myocardial scar and fibrosis. The 3D cine MRF exam is expected to have advantages over routine clinical imaging and existing rapid imaging methods in terms of (1) improved accuracy/reproducibility of quantitative tissue properties, (2) shorter exam times, (3) reduced operator dependence, and (4) high diagnostic accuracy for specific CM phenotypes. Technical validation of 3D cine MRF in healthy subjects will be performed in Aim 1, including development of cardiac/respiratory self-gating methods tailored for MRF and development of a “physics-informed” deep learning reconstruction for artifact reduction and scan acceleration. Aim 2 will compare image quality and quantitative measurements from 3D cine MRF to standard MRI methods in patients with established CM. Additionally, quantitative thresholds for objective detection of specific CM phenotypes will be determined. In Aim 3, tissue properties and synthetic images derived from 3D cine MRF for will be tested in a prospective cross-sectional study to evaluate diagnostic accuracy for differentiating (1) ischemic vs nonischemic CM and (2) nonischemic CM phenotypes, using a standard cardiac MRI protocol as reference. The overall expected outcome of this work is an ultrafast all-in-one MRI exam for CM detection and phenotyping that will streamline cardiac MRI exams and assess cardiac structure, function, and tissue properties using reproducible quantitative imaging.

NIH Research Projects · FY 2025 · 2022-06

Project Summary Magnetic Resonance Imaging (MRI) is one of the most important medical imaging modalities because of its ability to detect and characterize lesions throughout the body. However, access to MRI is severely limited by its expensive hardware, complex siting requirements and typically-qualitative images, which require highly skilled radiologists to interpret. This project proposes a fundamentally new way to encode MRI that could enable sub- stantially cheaper and more flexible quantitative MRI scanners. Today the overwhelming majority of MRI scans are encoded using two primary methods: B0 gradients and parallel imaging using an array of receiver coils. B0 gradients take up a significant fraction of the bore diam- eter; are loud and induce peripheral nerve stimulation, compromising patient comfort; they have relatively long switching times due to the high inductance of the coils; they require bulky cooling systems and customized am- plifiers; they are expensive, representing 25-30% of the cost of a clinical scanner; and they must be carefully designed and customized to a scanner's B0 magnet. B0 gradient encoding also suffers from spatial errors due to concomitant terms, which increase with decreasing B0 field strength and will limit the performance of emerging portable and low-cost MRI systems. Parallel imaging enables scan acceleration by differentiating signals across large spatial distances, but cannot encode complete images on its own. While some have proposed a third class of encoding methods using radiofrequency transmit (B1+) gradients, none of the methods described to date have been translated into clinical use because of practical limits on their performance, stringent hardware requirements and lack of flexibility in image contrast. This project will develop and validate a fourth, fundamentally new way to encode MRI based on parallel transmission using B1+-selective pulses produced by wireless RF coil units with on-coil amplifiers that perform RF transmission and reception, combined with an acquisition and reconstruction process inspired by MR Finger- printing (MRF). This new method, Selective Encoding through Nutation and Fingerprinting (SENF), completely eliminates the need for B0 gradients and is compatible with a wide range of magnet designs and flexible ac- quisition strategies. Unlike previous B1+ imaging methods, SENF places no strict spatial variation requirements on the RF gradient fields, which enables flexible system design, and the same coils can be used for spatial en- coding and signal reception. Furthermore, instead of suffering from errors due to complex spin dynamics during RF encoding, SENF leverages those dynamics to its advantage to differentiate quantitative tissue parameters. Successful completion of this project will enable a new generation of cheaper, more accessible, more modular, and lower-maintenance MRI scanners with quantitative outputs that can be more directly related to disease and tissue states.

NIH Research Projects · FY 2026 · 2022-05

REVISED PROJECT SUMMARY In this project titled “Collision of Alzheimer’s disease and COVID-19 pandemic in the United States: risks, outcomes, disparities and treatments”, we propose to: 1) examine COVID-19 risk in patients with Alzheimer’s disease (AD) and follow its evolution over time; 2) examine new onset of AD in COVID-19 survivors and follow these changes over time; 3) characterize long-haul neuropsychiatric Post-Acute Sequelae of COVID-19 (Neuro-PASC) in COVID-19 survivors with AD and follow these changes over time; and 4) investigate therapeutic effects of Tumor Necrosis Factor (TNF) blockers on overall mortality and long-haul Neuro-PASC in COVID-19 survivors with AD. To answer these questions, we will utilize a US nation-wide, real-time database of de-identified electronic health records (EHRs) of 80 million patients (4.8 million confirmed COVID-19 cases). Our study will primarily focus on over 17 million senior patients age  65 years, including more than 1 million COVID-19 cases, 265,818 patients diagnosed with AD, and 22,518 patients with both COVID-19 and AD (as of May 20, 2021). Our study will establish methods for real-time analysis of EHRs of vulnerable aging population of tens of millions across the USA in examining infectious diseases and their interaction with existing disorders such as AD, which will allow us to be better prepared should another pandemic overtake us. The significance of these studies therefore extends beyond the data at hand.

NIH Research Projects · FY 2026 · 2022-05

Project Summary/Abstract __________________________________________________________ Two-thirds (67%) of family caregivers to persons living with Alzheimer’s disease and related dementias (AD/ADRD) provide complex care tasks, such as medical/nursing tasks (e.g., managing medications, transferring from bed to chair, managing swallowing difficulties). Yet, only 53% of AD/ADRD caregivers receive any training to prepare them to conduct complex care. Consequently, AD/ADRD caregivers experience high levels of worry about making a mistake. Most responses to caregivers’ need for complex care training are not specific to AD/ADRD caregivers, though complex care is exponentially more challenging in the context of AD/ADRD. Provision of complex care to this population is complicated by the presence of behavioral symptoms of dementia (BPSD), difficulty with communication due to cognitive changes, and greater likelihood of multimorbidity than found amongst cognitively intact older adult care recipients. Another limitation of current responses to caregivers’ need for complex care training is current resources do not fully integrate principles of psychoeducation known to be effective at improving caregiver self-efficacy. High levels of self-efficacy, a person’s belief in their ability to accomplish a specific task, are associated with more positive perceptions of caregiving (e.g., meaningfulness), while low self-efficacy contributes to emotional distress, including depression. To build caregivers’ self-efficacy in the performance on complex care, Learning Skills Together (LST) was developed in 2017 at the UT Health San Antonio Caring for the Caregiver program by a multidisciplinary team with expertise in nursing, occupational therapy, speech-language pathology, nutrition, dental hygiene, and gerontology. In its most recent rendition, LST was delivered online over 4 synchronous videoconferencing sessions and program content integrated principles of Self-Efficacy Theory, such as peer-learning, modeling, and assignments so caregivers could practice skills and access feedback. In a single-arm pre- and post-test pilot study of LST, we observed statistically significant increases in self-efficacy at 4-weeks post-intervention (p=0.003). Near significant effects persisted 8-weeks postintervention (p=0.057). To rigorously test the efficacy of participation in LST on caregiver self-efficacy, we propose to test the hypothesis that caregivers to persons living with mid-stage AD/ADRD who participate in LST will report greater improvements in self-efficacy compared to a randomized active control group (N=200). We will also test for secondary outcomes we anticipate will be affected by improvements in self-efficacy, including caregiver depression and appraisal of BPSD. Subgroup analyses will be conducted according to caregiver characteristics, such as sex, to examine differences in intervention effects. If self-efficacy is demonstrated in this proposed study, the next step will be to examine effectiveness when delivering this program in community settings (e.g., Area Agencies on Aging).

NIH Research Projects · FY 2026 · 2022-05

The RNA-binding nuclear protein TDP-43 mislocalizes to the cytoplasm and aggregates in Frontotemporal lobar degeneration (FTLD-TDP variant), Amyotrophic Lateral Sclerosis (ALS), and >50% of late-onset Alzheimer’s disease (AD). Abnormal TDP-43 mislocalization and accumulation is associated with endoplasmic reticulum (ER) stress, synaptic dysfunction, and cognitive and motor impairments. While TDP-43 undergoes different post-translational modifications including phosphorylation, poly ADP-ribosylation, oxidation, acetylation, sumoylation, and ubiquitination, ubiquitination is a final key modification required for the turnover of TDP-43 via the ubiquitin-proteasome system and autophagy-lysosome pathways. TDP-43 is ubiquitinated by E3 ligases Parkin, PJA1, and Znf179. However, the role of deubiquitinases (DUBs) in the regulation of TDP-43 function, turnover, proteinopathy, and toxicity is poorly understood. The human genome encodes ~90 DUBs. Ubiquitin specific peptidases (USPs) are the largest family of DUBs comprising ~50 members in humans. Of these, 27 are expressed in the CNS. Our results from an unbiased screen of CNS-expressed DUBs identified USP19 as a major TDP-43 DUB, a positive regulator of TDP-43 stability, and a promising candidate for further study. Specifically, preliminary studies indicate that USP19, a DUB elevated during aging and in brains of FTLD-TDP patients, acts to increase TDP-43 stability/aggregation and participates in TDP-43-induced ER stress. By taking advantage of mouse models and human postmortem tissues together with molecular, cell biological, imaging, biochemical, proteomics, electrophysiological, behavioral, viral, histochemical, and recombinant protein toolsets, this proposal will 1. validate the role of USP19 in TDP-43 pathogenesis and associated phenotypes vivo, and 2. determine the mechanistic basis of USP19 in TDP-43 deubiquitination, stability, aggregation, and toxicity in genetically modified neurons and in vitro systems. Successful conclusion of these studies will determine the significant contribution of USP19 and its DUB activity to TDP-43 pathogenesis in humans and mice. Moreover, these results will provide novel mechanistic insights to USP19 DUB activity in concert with TDP-43 in ER stress and neurotoxicity. Together, these studies will enable the pursuit of a potential therapeutic direction of targeting USP19-mediated mechanisms to mitigate TDP-43 pathology and toxicity.

NIH Research Projects · FY 2026 · 2022-04

The microbiome and mucosal immunity in cervical cancer disparities African American women living in the United States continue to experience an undue higher burden of cervical cancer and a >2 times higher mortality rate than European American women. This survival disparity persists after accounting for socioeconomic status and disease stage. The presence of high-risk human papillomavirus (HPV) is the major cause of cervical cancer and cervical intraepithelial neoplasia. Prophylactic vaccines are available against the most carcinogenic types and are highly effective, but disparities persist and the number of people receiving the vaccine remains suboptimal especially for African American women, and the vaccine is ineffective for the 40% of female US population already infected with genital HPV. Treatment options for advanced stages are limited, and metastatic cancer is incurable. New therapeutic approaches are therefore needed but the mucosal mechanisms contributing to disease pathogenesis in African American women are not well understood. Therefore, identifying mucosal processes and/or mediators which modify HPV persistence vs clearance and progression vs remission of cervical neoplasia could lead to new or improved treatments and better CC outcomes for women. In this proposal we will investigate the contribution of the microbiome and mucosal immunity in the female genital tract to health disparities in cervical cancer in African American women. This is built upon considerable data from our lab and others that show that African American women have higher proportion of vaginal microbial dysbiosis; that vaginal microbial dysbiosis is linked to pro-inflammatory and cancer pathways in cervical mucosa; that these bacteria produce metabolites that are linked to an immunosuppressive phenotype; that dysbiotic bacteria can induce cancer pathways in vitro; and cervical cancer tissue gene expression data from African American women show increased activation inflammatory pathways compared to European American women. In this concept we will utilize state-of-the-art systems biology techniques, including metagenomics, metaproteomics, metabolomics, single cell RNA sequencing and flow cytometry to study vaginal mucosal biology in prospective studies of African American women, coupled with functional studies in PV-associated cervical cancer mouse models, to better understand these relationships.

NIH Research Projects · FY 2026 · 2022-04

Obesity is a chronic progressive disease that leads to the development of heart disease, stroke, and type 2 diabetes, which are among the top ten leading causes of death in the United States. The initiation and progression of obesity-related diseases is strongly associated with chronic low-grade inflammation, and the NLRP3 inflammasome is a key sensor that instigates inflammation in obesity. Targeting NLRP3 inflammasome-mediated inflammation should curb or prevent the disease progression and thus holds therapeutic promise to combat obesity-related diseases. However, effective and safe strategies that specifically inhibit the NLRP3 inflammasome in obesity have not been developed for patient treatment. As such, the long- term goal of our research is to develop a new dietary strategy or therapeutic modality to suppress NLRP3 inflammasome activity and inflammation in obesity and obesity-related diseases. As an initial step, this project will define the role of chive-derived exosome-like nanoparticles (C-ELNs) in suppressing inflammation in obesity. Our pilot studies found that C-ELNs strongly inhibited NLRP3 inflammasome activation in primary macrophages. One of their bioactive molecules, 1,2-dilinoleoyl-sn-glcyero-3-phosphocholine, was identified as an inhibitor of the NLRP3 inflammasome. Oral administration of C-ELNs, started concomitantly with high-fat diet feeding, reduced NLRP3 inflammasome activity and improved metabolic health in the C57BL/6J mice. Building on our preliminary work, this project will test the central hypothesis that C-ELNs contain active biomolecules that inhibit NLRP3 inflammasome activity and ameliorate inflammation in obesity. This hypothesis will be tested through two specific aims: 1) identify active biomolecules in C-ELNs that inhibit NLRP3 inflammasome activity and 2) define the role of C-ELNs and their active biomolecules in suppressing inflammation in obesity. Successful completion of the proposed research be the first step toward the translation of C-ELNs and active biomolecules into an intervention to suppress NLRP3 inflammasome activity and inflammation in obesity. Utilizing dietary ELNs to target the NLRP3 inflammasome is an innovative approach. The unique features of dietary ELNs, including tissue bioavailability, bioactivity, and biomolecule protection and delivery, as well as their abundance in edible plants confer upon them high translational potential.

NIH Research Projects · FY 2025 · 2022-04

Although in cystic fibrosis (CF) patients, respiratory and digestive disease is the primary source of morbidity and mortality there are many other clinically relevant symptoms such as depression and anxiety, and poor sleep quality, including sleep disturbances, and altered sleep patterns. Sleep disturbances experienced by individuals with CF are consistent with circadian rhythm (CR) phase delays. The clinical relevance of CR and sleep regulation in CF can be seen in studies that demonstrate CF patients with poor sleep quality have more severe lung disease and poorer outcomes over time. Whether CR disruption in CF is a direct effect of impaired CFTR function or a secondary manifestation of disease progression is currently unclear. Also unclear is the efficacy of modulatory therapy in CF in reversing these phenotypes. We have recently published that CR gene expression is altered in a CF mouse model suggesting that CR dysregulation is a primary manifestation of CF. Mechanistically, we have previously reported alterations in microtubule regulation in CF cells. These findings lead to the hypothesis that CR regulation in CF is altered due microtubule instability and consequent reductions in melatonin production. Microtubules have been suggested to play an important role in CR, and we have previously demonstrated that CF cells display reduced acetylation and slower microtubule formation rates. We have also recently published that depletion of a microtubule modulating protein (tubulin polymerization promoting protein, TPPP) from mice replicates CF-like CR disruptions that support a role of microtubules in CF phenotypes. Preliminary data demonstrate that CF mice produce reduced levels of melatonin, a key CR regulator and known regulator of microtubule stability. These data suggest melatonin and/or microtubule targeted compounds as possible therapeutic interventions that can augment modulator therapy or allow an alternative approach to address CR-related phenotypes in CF patients. The goals of the study are to determine the role of CFTR in CR regulation and if CFTR correction influences CR gene expression and related behavioral outcomes. We also will strive to understand the cellular mechanisms involved in these regulatory relationships that can be therapeutically targeted. To achieve these goals, the following specific aims will be studied: Aim 1. To determine the CFTR- dependency of CR regulation and the efficacy of highly-effective CFTR modulators in reversing CF-related CR phenotypes. Aim 2. To identify mechanisms of CR dysregulation in CF. Aim 3. To determine the effect of CFTR modulators on circadian rhythm and to determine melatonin production in children and adolescents with CF.

NIH Research Projects · FY 2025 · 2022-04

Abstract Ulcerative colitis (UC) is a chronic form of inflammatory bowel disease (IBD) with no cure. Current treatment strategies offer only partial remission, with many patients remaining refractory to treatment, and carry risks of significant adverse events including serious infections and cancer. Thus, an improved understanding of inflammatory signaling pathways and identification of novel preclinical mechanisms are critical challenges in IBD research. Signaling downstream of the proinflammatory cytokine tumor necrosis factor (TNF) and toll like receptor (TLR) pathways play major roles in IBD pathogenesis. In previous work, we discovered that the RNA binding protein Sam68 is required for both TNF- and TLR-induced activation of the transcription factor NF-κB, a master regulator of inflammation, suggesting that Sam68 contributes to NF-κB-dependent inflammation. Our new preliminary results show that Sam68 is prominently expressed in human and murine intestinal epithelial cells (IEC); Sam68 knockout (KO) mice are significantly protected from dextran sulfate sodium (DSS)- and oxazolone-induced colitis; and Sam68 protein is significantly elevated in inflamed colons of UC patients. Based on this, we hypothesize that Sam68 is a critical mediator of inflammation in IECs, and targeting IEC Sam68 will provide a novel therapeutic strategy in UC via dual inhibition of TNF- and TLR-mediated inflammatory pathways. We propose to study the molecular mechanisms of IEC-specific Sam68 signaling in human and experimental murine colitis using cutting edge tools such as 3D colonoids derived from UC patient colonocytes and novel IEC-specific Sam68 conditional KO (cKO) mice. Aim 1 will delineate molecular mechanisms of IEC-specific Sam68 signaling in TNF- and TLR-induced inflammatory signaling and identify the structural domains and posttranslational modifications of Sam68 required for its inflammatory functions. Aim 2 will study the in vivo role(s) of Sam68 in primary IECs under homeostatic and inflammatory conditions, using Sam68-cKO mice challenged with DSS and oxazolone as experimental colitis models, and using a spontaneous colitis model, Sam68-cKO / Wiscott Aldrich Syndrome Protein (WASP) KO double KO mice. This aim will utilize 3D colonoids prepared from wild type and Sam68-KO mice to study role of Sam68 in IEC proliferation, permeability and inflammatory signaling to study the homeostatic role of Sam68. Aim 3 will delineate the proinflammatory role of Sam68 in UC patients. This aim will study IEC-specific growth, proliferation, apoptosis, and expression of TJ proteins, and activation of proinflammatory signaling in IEC's and mucosal immune cells using co-culture assays of control and patient derived 3D intestinal organoids and immune cells. Successful completion of this study will fill critical knowledge gaps in understanding novel mechanisms underlying TNF- and TLR-dependent inflammatory signaling in UC and pave the way for the development of novel therapeutics targeting Sam68 to treat UC. Moreover, this study will also serve as the basis to explore the role of Sam68 in other TNF- and TLR-dependent chronic inflammatory diseases as well.

NIH Research Projects · FY 2026 · 2022-04

This new D43 application proposes a training program entitled “Strengthening Research Capacity in Innovative Global Health Technologies for Non-Communicable Diseases in Uganda” (SIGHT). This program will build on the extensive 32-year training record of the Uganda-CWRU Research Collaboration (UCRC), a partnership between Case Western Reserve University and Makerere University The burden of non-communicable diseases and disorders (NCD) is growing rapidly in low- and middle- income countries (LMIC) and sub-Saharan Africa (SSA) and Uganda particularly. Developing capacity for biomedical research in LMICs is necessary for managing these new healthcare challenges, and encouraging progress is being made. We assert that building capacity in particular for biomedical engineering (BME) research is critical to development of much-needed screening, diagnostic, and therapeutic technology relevant to the context of LMICs. Training programs to build this capacity are lacking. We have identified three broad categories of NCDs to target in the SIGHT program that will particularly benefit from building capacity for technology innovation: cardiovascular disease, blood disorders, and chronic movement disorders. These target areas have been identified as critical public health needs by program leaders and other stakeholders. We have also identified four areas of technology focus as highest priorities for building capacity in BME research expertise at MU in Uganda. These are: biomaterials and drug delivery, point-of-care diagnostics, biomedical imaging, and data analytics and artificial intelligence. These technology tools are cross-cutting in their potential to address high-priority NCD healthcare needs. The SIGHT program has been designed with the long-term goal of building and strengthening the capacity of academic, public, private, and NGO institutions in Uganda to conduct biomedical engineering research, train biomedical engineers up to a PhD level and grow a local biomedical engineering industry based on local needs. SIGHT aims to contribute to this goal by: Aim 1: Training Ugandan students in the BME Ph.D. program at CWRU (6 within the proposed project period) Aim 2: Supporting research projects for M.S. students at MU in BME-adjacent fields (approx. 10 per year). Aim 3: Providing opportunities for MU investigators to enhance their research experience and expertise. Expected outcomes of the SIGHT program include: ? Ph.D. researchers prepared to fill faculty and research positions at research institutions in Uganda. ? M.S. graduates who are prepared to fill research positions or go on to Ph.D. training. ? MU investigators who are better prepared to carry out high quality BME research. ? A robust network of research collaborations between BME and clinical investigators at CWRU and MU. ? Increased research output, including publications in peer reviewed journals and grant proposals.

NIH Research Projects · FY 2025 · 2022-04

Background. This proposal, which is submitted in response to RFA-AI-21-021 “Understanding Post-Transcriptional Regulation of Intact and Defective HIV RNA”. N6-methyladenosine (m6A), is the most common RNA modification and is known to regulate RNA stability, splicing and nuclear export. m6A modification of HIV transcripts is crucial for the early stages of HIV infection during acute infection of primary T cells, but it is an open question whether m6A modification controls HIV latency and reactivation in ART-suppressed patients. Our goal. Our multidisciplinary team has extensive experience in studies of HIV latency and reactivation in patients and in reliable primary cell models, studies of RNA m6A modification, and cutting-edge technologies such as NGS sequencing and scRNA-seq analysis. To overcome the challenge of measuring m6A in RNA recovered from the extremely low numbers of HIV+ cells present in patient samples, we will develop a sensitive next-generation sequencing assay for the profiling and quantification of m6A modification in different HIV transcripts from patient samples. This assay, which we call MeRIP-EDITS combines methylated RNA immunoprecipation with the EDITS assay, which has been used in multiple clinical studies to measure the inducible HIV reservoir. We will use the MeRIP-EDITS assay to characterize m6A modification of different HIV transcripts at different reactivation kinetic points of latent HIV and examine changes of the m6A pathway during HIV latency and reactivation. In parallel we will perform mechanistic studies on the m6A pathway using the QUECEL primary cell model of HIV latency. We will use the model to develop a sensitive nanopore RNA-sequencing assay which can subsequently be applied to patient samples. We will also inhibit the activity of the m6A writer METTL3 and the erasers FTO and ALKBH5 by knocking out the expression of these genes by using the CRISPR gene editing technology. High resolution mRNA FISH experiments, which distinguish between spliced and partially spliced HIV mRNA transcripts will be used to study the colocalization of m6A readers and HIV mRNAs. How will we advance the field? Demonstration of a central role of m6A in the control of HIV latency would immediately suggest pharmacological strategies to incorporate into HIV cure regimens. To date, it has been impossible to efficiently reverse HIV latency using agents that are designed for “kick and kill” strategies for an HIV cure. Using the sensitive assays described above, we will evaluate the impact of inhibitors of m6A erasers as part of a “kick and kill” strategy for HIV latency reversal. As a complementary approach we will also evaluate whether inhibitors of m6A writers can inhibit HIV reactivation and lead to long term silencing, as part of a “block and lock” strategy.

NIH Research Projects · FY 2025 · 2022-02

Project Summary Tyrosine kinase inhibitors (TKIs) are important therapeutic agents to treat various cancers. However, any agent has a tradeoff between efficacy and on or off target deleterious effects. This notion became evident in the treatment of chronic myelogenous leukemia (CML) when a potent and broadly inhibitory tyrosine kinase inhibitor, ponatinib (Iclusig, Ariad Pharmaceuticals, now Takeda) was recognized to have a 31% incidence of cardiovascular (CV) events of which 21% overall were significant adverse events (SAEs). In a Phase II trial (PACE) at 4 yrs. the incidence of arterial occlusive events was 26% (myocardial infarction 14%, stroke 11%, and limb ischemia 11% - some patients have more than 1 organ event). Ponatinib (poni) is one of 5 TKIs approved for the treatment of CML We have created a murine model to examine the effects of TKIs on blood coagulation, vascular, and platelet function. In aged mice treated with the various TKIs under steady-state conditions, ponatinib, unlike imatinib, demonstrated an increased risk of arterial and venous thrombosis. Poni treatment leads to decreased arterial occlusion times, larger venous clots and generates hyperactive platelets - features that contribute to heightened thrombosis. Our laboratory has identified key mechanisms underlying the prothrombotic phenotype of poni. First, poni-treated mice have increased vessel wall reactive oxygen species (ROS), apoptosis, and inflammatory vascular lymphocyte infiltrates that expresses coagulation factors V and VIII. Second, platelets from poni-treated mice are hyperactive to in response to collagen. Additionally, we have determined that pioglitazone (pio), a PPAR agonist, when given with poni normalizes the vessel wall inflammation and platelet hyperactivity to correct murine thrombosis risk The overall hypothesis of this application is that poni-associated thrombosis results from immune cell vascular inflammation expressing prothrombotic genes and altered platelet signaling resulting in platelet hyperreactivity. Poni treatment has identified a novel mechanism of prothrombotic vascular dysfunction by which vascular infiltrating lymphocytes express coagulation enzymes FV and FVIII potentially to contribute to thrombosis. At therapeutic dosing in man, poni inhibits p-LynY507, a negative regulator of activated GPVI, in both unstimulated and activated platelets with little effect on p-LynY396 and p-SykY352, suggesting that these platelets may be more reactive. In fact, poni-treated mice have platelets that react to lower concentrations of CRP. These defects are genetically and functionally corrected by pio’s genomic and non-genomic PPAR agonism. The specific aims of the proposal are as follows: The specific aims of the proposal are as follows: 1) Determine the mechanism of ponatinib- and other TKI- induced vascular inflammation 2) Identify the mechanisms of poni-induced platelet hyperactivation. These studies will determine the mechanisms of poni and other TKI effects on vessel wall and platelets that lead to cardiovascular events. They present a pre-clinical model for poni-associated thrombosis and correction with pio, a PPAR agonist. Last, they will serve as a paradigm for CVD assessment for TKIs in general.

NIH Research Projects · FY 2026 · 2022-02

Project Summary PM2.5 is a leading global risk factor for morbidity and mortality and is the leading environmental factor implicated in cardiovascular disease. Recent data suggest that PM2.5 and/or traffic-related air pollutants may play a role in chronic diseases such as Type 2 diabetes mellitus, that may in turn confer susceptibility to cardiovascular disease and has been the topic of multiple systematic reviews and meta-analysis by us and others. In a series of studies in animals and humans, we have provided convincing evidence linking PM2.5 to pathways playing an etiologic role in cardiometabolic disease. Our findings and reviews have fundamentally shaped the understanding of the effects of PM2.5 and its association with increased risk of cardiometabolic disease. In particular, our group has demonstrated pro-atherogenic effects of chronic PM2.5 exposure, through ROS and inflammation dependent pathways. Although our understanding of pathways contributing to progression of disease has been dramatically enhanced, the mechanisms involved in the resolution of inflammation and an understanding of how dysregulation of resolution pathways contributes to disease progression has been limited. Identification of these mechanisms could pave the way for new opportunities to target non only air pollution mediated cardiovascular risk but other pervasive health risks. Through the implementation of three specific aims this proposal seeks to: 1) characterize air pollution-induced changes in inflammation resolution pathways; 2) determine to what extent these pathways overlap with dietary manipulation; 3) identify potential intervention strategies to promote upregulation of conserved resolution pathways in response to air pollution-induced inflammation.

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY Myelin degeneration and white matter loss that result from oligodendrocyte (OL) death are early events in Alzheimer’s disease (AD) that lead to cognitive deficits and correlate with disease status. The loss of OLs, accompanied by a reduction of myelin density, axonal loss, and astrogliosis, are major changes in white matter that occur in the brains in both AD patients and animal models of AD. OLs are the most abundant glial cell type in the brain but the least studied cell population in the context of neurodegeneration, despite their vital role in myelin maintenance and neuronal support. Recent genome-wide association studies and large-scale single- cell transcriptomics of AD patient brains emphasized the crucial role of OLs in the development of AD. The underlying mechanisms of OL dysfunction and its contribution to the initiation and progression of AD remain unknown. Our recent work reports, for the first time, that mature OLs in AD patients and AD mice exhibit NLRP3 inflammasome-associated inflammatory injury, concomitant with demyelination and axonal degeneration. Unbiased proteomic analysis further suggests that the hexokinase 1 (HK1)-dependent glycolysis pathway is most suppressed in AD mouse white matter. Mature OLs rely heavily on glycolysis for energy production, even in the presence of oxygen. HK is the rate-limiting enzyme that initiates the first step of glycolysis by the phosphorylation of glucose. OLs specifically express a brain HK isoform, HK1. HK1 localizes to the mitochondrial outer membrane, and the dissociation of HK1 from mitochondria decreases its enzymatic activity, which is sufficient to inhibit glycolysis and induce NLRP3 inflammasome activation. We found that HK1 immunodensity and enzyme activity significantly decreased in OLs in AD patients and AD mice. In mature OLs in AD, the HK1 mitochondrial association is disrupted by overactivation of the mitochondrial fission protein dynamin-related protein 1 (Drp1), and Drp1 and HK1 synergistically elicit NLRP3 inflammasome activation and the release of interleukin-1β, triggering inflammation. The mature OL-specific heterozygous knockout of Drp1 in AD mice restores HK1-dependent glycolysis, abolishes NLRP3 inflammasome activation, corrects myelin loss, reduces neuroinflammation and axonal degeneration, and improves cognitive function in animals. These findings support the scientific premise of the proposed project that glycolytic deficiency in OLs, driven by the Drp1-HK1 molecular switch, induces OL metabolic dysregulation and inflammation and causes white matter degeneration, AD pathology, and cognitive impairment. Successful completion of the proposed studies will support the hypothesis that OL metabolic deficiency is a key pathological process that induces inflammation, demyelination, white matter loss, and AD-associated neuropathology and cognitive deficits. These studies are crucial to further reveal the role of the novel Drp1-HK1 OL pathway in AD and determine whether this pathological pathway is a plausible treatment target for AD.

NIH Research Projects · FY 2025 · 2022-02

PROJECT SUMMARY/ABSTRACT Non-alcoholic Fatty Liver Disease (NAFLD) is characterized by hepatocyte fat accumulation in the absence of alcoholic or viral etiologies, and is part of a spectrum of disease ranging from isolated steatosis to hepatocellular carcinoma (HCC). NAFLD is estimated to affect nearly one-quarter of the world's population. As the incidence of NAFLD-associated HCC has risen rapidly over the past few decades, it has become clear that NAFLD is an emerging public health crisis for which there is currently no approved pharmacologic intervention. This project will investigate the role of AKR1a1—a recently discovered protein denitrosylase—in the pathogenesis of NAFLD through interrogation of its role in modulating hepatic lipid metabolism via reversible S- nitrosylation of key lipogenic enzymes. Further, the potential for AKR1a1 as a novel therapeutic target to prevent NAFLD will be investigated. To this end, the study will employ a variety of techniques including: CRISPR-Cas9 genome editing and administration of small molecule inhibitors in dietary models of murine NAFLD; isotope tracing studies to interrogate whole-pathway and enzyme-specific effects on lipid metabolism; resin-assisted capture to assess endogenous S-nitrosylation of enzymes; and site-directed mutagenesis to determine the functional role of S-nitrosylation. Together, this study will provide novel insight into the regulation of hepatic lipid metabolism and the pathophysiology of NAFLD, and identify potential therapeutic targets.

NIH Research Projects · FY 2025 · 2022-01

Red blood cells (RBCs) play a vital role in gas transport—carrying O2 from the alveolar air to systemic tissues, and CO2 in the opposite direction. Their task is central to many diseases of major public-health relevance, in- cluding including heart failure, pulmonary disease (including COVID-19), vascular disease, and sepsis (hy- poperfusion). An important component in the movement of these gases within the body is the transport of these gases across of the plasma membrane (PM) of the RBCs. The dogma had been that all gases cross all mem- branes merely by dissolving in and diffusing through membrane lipids. However, challenging this dogma was the discovery of the first CO2 impermeable membranes, and the first evidence that a gas (CO2) moves through a membrane protein (the water channel aquaporin 1, AQP1). In human RBCs, aquaporin-1 (AQP1) and the Rh complex (including RhAG) account for 90% of membrane CO2 permeability. Preliminary data on O2-offloading from RBCs from knockout (KO) suggests that these two channels, together, are responsible for ~55% of O2 permeability (PM,O2). The addition of the membrane-impermeant inhibitor pCMBS to RBCs from the double- knockout (dKO) mouse reduces PM,O2 by ~90%. Aging mice appear to gradually undergo a decrease in PM,O2 that does not occur in dKOs. A surprising preliminary observation is that the knockout (KO) of one or both of these channels reduces maximal O2 uptake rate (V?O2 max) without decreasing—and, in fact, often increasing—running performance. This grant has two aims. Aim 1 is to determine the extent to which channels vs. lipid composition contribute to the rate of O2 offloading (kHbO2). One approach is to study aging wild-type (WT) vs. KO mice. Another is to examine mice with RBCs genetically depleted or replete in AE1, or depleted in MCT1. The third approach is to examine mice of disease models or widely different genetic background. In each case, the investigators will examine hematology, RBC size and shape, proteomics, lipidomics, and genomics. 3D macroscopic mathemati- cal modeling will play a central role in data interpretation. Finally, the investigators will use exercise protocols to to determine V?O2 max, critical speed, exercise economy, and speed of V?O2 kinetics. They will also examine cardio- vascular and muscle parameters. In Aim 2, the goal is to elucidate the molecular mechanism of O2 movement through AQP1, RhAG, and candidate O2 channels (e.g., AE1). The investigators will use an iterative approach, the first step of which involves identifying prioritizing missense single nucleotide polymorphisms (SNPs), as well as other mutations that come forward in Aim 1. The investigators will use a novel neutral buoyance assay to measure O2 uptake into oocytes and thereby assess these mutants channels. Molecular dynamics and molecular biophysics will complete the iteration before choosing additional laboratory mutation for analysis. The proposed research will reorganize thinking about O2 carriage by blood and could lead to therapies to improve exercise in patients with diminished exercise capacity.

NIH Research Projects · FY 2026 · 2021-12

SUMMARY Dominant FBN1 mutations cause Marfan syndrome, an inherited human connective tissue disorder affecting fibrillin-1 microfibrils and leading to thoracic aortic aneurysms with risk of aortic dissection and rupture. Fibrillin- 1 is a product of vascular smooth muscle cells (VSMC), which provides an important link in the mechanical continuum from the SMC contractile cytoskeleton to the extracellular matrix, in addition to providing a template for elastic fiber assembly. Reduced tissue fibrillin-1 content as a result of FBN1 haploinsufficiency is thought to be the mechanism underlying a significant proportion (over 1/3) of Marfan syndrome mutations. In recent work we found that ADAMTS6, a metalloprotease secreted by VSMC, cleaves both fibrillin-1 and fibrillin-2. The latter is produced primarily during the embryonic period and is thought to have a minor role in the aorta after birth. Analysis of a mouse Adamts6 null mutant, which does not survive past birth, shows an accumulation of both fibrillin-1 and fibrillin-2, with major skeletal and cardiac defects we have genetically attributed to fibrillin-2 accumulation. Thus, ADAMTS6 appears to be a major protease regulating fibrillin microfibril turnover. This provides a compelling rationale for targeting ADAMTS6 in Marfan syndrome in a novel disease- modifying approach. Based on these findings, the overarching hypothesis of this proposal is that ADAMTS6 inactivation in vascular smooth muscle cells postnatally will protect aortic fibrillin-1 microfibrils from proteolytic turnover, thus increasing microfibril abundance and mitigating aortic aneurysm growth and dissection in Marfan syndrome. In Aim 1, we will use a new Adamts6 conditional mutant to test this hypothesis through conditional deletion of Adamts6 in VSMCs in a well-characterized mouse model of severe Marfan syndrome that reliably progresses to dissection and rupture. In Aim 2, we will define the intermolecular interaction of fibrillin-1 and ADAMTS6 to identify the major molecular determinants of proteolysis. In vitro microfibril assembly will be used to test the impact of blocking ADAMTS6-fibrillin interactions. These experiments will inform future approaches for protecting microfibrils from ADAMTS6-mediated turnover. Impact: A disease-modifying approach for Marfan syndrome does not exists, and non-surgical options have not been wholly effective in preventing dissection. These aims leverage our initial discovery that ADAMTS6 cleaves fibrillin-1 for continued investigations intended to drive development of an ADAMTS6 blockade-based disease- modifying approach for Marfan syndrome. Specifically, the disease mechanism in many patients is reduction of fibrillin-1 microfibrils and we aim to enhance microfibril abundance by protecting them from breakdown. Together the aims provide a proof of principle for a possible disease-modifying therapy (Aim 1) and the basis for interfering with ADAMTS6 cleavage of fibrillin-1 (Aim 2). The work proposed herein also addresses fundamental questions of how fibrillin-1 is turned over in the vascular wall.

NIH Research Projects · FY 2025 · 2021-09

Abstract A seizure is an abnormal neural activity in the brain and the main characteristic of epilepsy. About one-third of the patients have seizures resistant to anti-epileptic medication and therefore, an alternative treatment is necessary. Recent studies show that neurons can be recruited and seizures can propagate by a mechanism known as electric field coupling. Our laboratory has shown that canceling the electric field in the extracellular space can prevent neural propagation and recruitment. We now propose to develop a novel neurotechnology that not only can control the propagation of seizures but prevent the seizure from being generated by controlling the extracellular electric field. With a combined methodology of in-vitro electrophysiology with state-of-the-art neural imaging, computer modeling and in-vivo preparations in rodents, we will implement the seizure control system and study its mechanism with four specific aims: 1) Prevent neuronal synchronization in hyperexcitable neural tissue, 2) Develop a novel technology of extracellular voltage clamp to control seizures in-vivo, 3) Determine the mechanisms of seizure control by an extracellular voltage clamp system, 4) Determine the role of the extracellular space in the clamping efficiency. Current electrical responsive control therapy detects a seizure and then applies stimulation. The proposed neurotechnology will prevent the synchronization between neurons to stop the generation of seizures before they arise at very low current amplitude thereby providing a novel therapeutic paradigm for treating epilepsy.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Alzheimer's disease (AD) and AD-related dementias (ADRD) such as Lewy body dementia (LBD) are all associated with deposition of misfolded protein aggregates in the brain including tau in AD and non-AD tauopathies, and α-synuclein (αSyn) in LBD. Currently, a definite diagnosis of these disorders relies on the histological and biochemical examination of the brain for the misfolded proteins. Development of reliable and sensitive assays for these misfolded proteins in easily accessible peripheral specimens is critical for early or differential diagnosis, determination of disease severity, and evaluation of therapeutic efficacy in clinical trials. Interestingly, brain tau and αSyn aggregates exhibit prion-like aggregation seeding activity, which can be specifically detected by two highly sensitive amplification assays including real-time quaking-induced conversion (RT-QuIC) and protein misfolding cyclic amplification (PMCA). They have been proved to be highly sensitive for detection of misfolded proteins in the brain and/or cerebrospinal fluid in prion disease (PrD), AD, or PD (Atarashi et al., 2011; Peden et al., 2012; Foutz et al., 2017; Orrú et al., 2015; Saijo et al., 2017; Kraus et al., 2018). Using RT-QuIC/PMCA, we were able to detect prion and αSyn aggregates in the skin of individuals with PrD or PD (Orrú et al., 2017; Wang et al., 2019; 2020). Remarkably, our preliminary results have shown that prions-like tau- seeding activity is detectable by RT-QuIC and PMCA in skin of AD patients but not in normal controls. Thus, we hypothesize that skin tau-seeding activity detected by RT-QuIC and PMCA is a novel biomarker for diagnosing, characterizing, and predicting outcomes of AD and non-AD tauopathies and for differentiating AD from LBD. To test this hypothesis, the following four Aims will be pursued: (1) Establish the tau-seeding activity in autopsied skin samples as a biomarker for POSTMORTEM diagnosis and characterization of AD using RT-QuIC/PMCA assays; (2) Assess skin tau-seeding activity as a biomarker for PREMORTEM diagnosis, characterization, and predicting clinical outcomes of AD; (3) Determine skin tau-seeding activity as a biomarker for differentiating AD from non-AD tauopathies, and from LBD, a common ADRD; and (4) Determine whether skin tau-seeding activity is detectable at an asymptomatic stage by RT-QuIC/PMCA in animal models of AD tauopathies. We believe that the successful implementation of this project will develop RT-QuIC/PMCA assays of skin tau-seeding activity as a biomarker for diagnostic testing and evaluating clinical trials across AD, non-AD tauopathies, and LBD.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT / PROJECT SUMMARY People living with multiple chronic conditions (MCC) consume a high proportion of health care resources but are not well-served by one-disease-at-a-time scientific evidence and health care delivery systems. Primary care, focused on the whole person in their family and community context, has great but under-informed potential to integrate the care of people with MCC, particularly community health centers (CHCs) that focus on health disparity populations. The COVID-19 pandemic and resulting societal and health care system responses have uncovered tremendous care disparities, while also stimulating innovations with great potential to foster integrated personalized primary care for people with MCC in an otherwise fragmented, impersonal, inequitable health care system. In particular, telehealth visits in the context of ongoing person- focused relationships, integrated mental health services, and outreach to high-risk patients with MCC provide hope for advancing the equity and quality of care for health disparity populations. We have a unique opportunity to study the effects of the pandemic in a national network of 926 community health centers serving >2.6 million patients from health disparity populations. The proposed study will: 1) Identify evolving changes in health care delivery to people with MCC in community health centers in response to the continuing COVID-19 pandemic and its aftermath; 2) Assess the impact of these practice changes on care quality for people living with MCC; 3) Identify promising emerging strategies to improve health care for people living with MCC. We will conduct a time series analysis of practice changes and associated patient outcomes in response to the COVID-19 pandemic. Monthly analyses will examine changes in practice processes, patient outcomes, and workforce stability for health disparity populations and for people with and without multiple chronic conditions. For a five-year period going back to January 2020 ― the month of the first known COVID- 19 case in the US and the start of responding public health and practice changes ― we will analyze and publicly report, on a monthly basis, practice changes and patient outcomes, to rapidly inform decision making about primary care of health disparity populations with MCC. Subgroup analyses will examine differences across 26 states that have had different societal and policy responses to the pandemic. The findings from these quantitative analyses of millions of patients being seen in hundreds of community health centers will be used to identify a purposive sample of exemplars for in-depth case studies that put a face on the quantitative findings and identify and characterize practice innovations that show promise in reducing health care disparities and improving care for people living with MCC. Study findings will generate vital new knowledge on the effect of a pandemic on the quality and equity of care provided to people MCC and will inform efforts to improve health care equity after a natural disaster.

NIH Research Projects · FY 2025 · 2021-09

Unveiling mechanisms of neural stimulation technologies is an important goal of the Brain Initiative (RFA-NS-20-006). Transcranial electric stimulation (TES) is a non-invasive neuromodulation technique to provide wide-range effects on seizure control, behaviors, and cognition by generating weak electric fields in the brain. It is still an open question of how these weak electric fields can interact effectively with neural activity, and the mechanism of action of TES is still unknown. Our laboratory has recently found that ephaptic coupling (non-synaptic neural coupling by electric fields) plays a significant role in neural recruitment and could possibly explain the TES-induced effects. We propose to test the hypothesis that ephaptic coupling is the mechanism of action of TES. With the combination of in-vitro novel voltage imaging techniques and optogenetic stimulation in the transgenic mice and in-vivo electrophysiological experiments, we will study the links between the ephaptic coupling and three known TES-induced effects in the following independent specific aims: 1) Investigate the role of ephaptic coupling in TES-induced propagation speed modulation of interictal spikes. 2) Determine the role of ephaptic coupling in TES-induced seizure suppression. 3) Study the role of ephaptic coupling in TES-induced effects on neural oscillations. This proposal, if successful, will provide a novel insight into TES-induced effects, reveal the mechanism of action of TES, and open the door to the development of improved TES protocols with improved efficiency for this non-synaptic therapeutic modality.

NIH Research Projects · FY 2025 · 2021-09

Project Summary/Abstract Kaposi sarcoma (KS) remains one of the most common malignancies in people living with HIV/AIDS worldwide. Kaposi sarcoma herpesvirus (KSHV), also called human herpesvirus-8 (HHV-8), is the causal agent for KS. The oral mucosa is the first target of KSHV infection once the virus is in the oral cavity. However, the initial infection process of the mucosa by KSHV has never been studied, mainly due to lack of an in vitro model to recapitulate the viral infection in vivo and nonexistence of an animal model for KSHV infection. We have created the 3- dimentional (3-D) organotypic culture as an oral mucosal mimic resembling a stratified oral mucosa with the potential for histological assessment of the KSHV infection/transmission process. Our studies show that KSHV infection in the 3-D oral mucosa model is enhanced by saliva extracellular vesicles (EVs), particularly exosomes, from HIV patients. Exosomes purified from the saliva of HIV patients or from culture media of HIV-infected T cells contain similar HIV-specific cargos, including HIV TAR RNA. KSHV entry into target cells and its infectivity are increased by HIV-positive saliva exosomes in primary and immortalized human oral epithelial cells. Further, KSHV infection and transmission to the suprabasal cells in the 3-D organotypic culture are enhanced by HIV- positive exosomes. Increased KSHV infectivity by HIV-positive exosomes is attributed to the HIV TAR RNA within the vesicles and epidermal growth factor receptor (EGFR) of host cells. Inhibition of EGFR by Cetuximab, a monoclonal antibody to EGFR, blocks KSHV infection enhanced by HIV-positive exosomes. Taken together, these data lead us to hypothesize that HIV-positive exosomes promote KSHV infection and transmission through the oral route and are responsible for increased incidence of KSHV infection in people living with HIV. To test this hypothesis, we plan to 1) assess KSHV transmission and infection through the oral route with the 3-D cultures of oral mucosal and tonsil tissues incorporated with peripheral blood mononuclear cells (PBMCs) as well as the epithelium-endothelium model. KSHV infection and transmission through the epithelial barrier to immune and endothelial cells in response to HIV+ saliva exosomes will be assessed; 2) delineate the mechanism by which HIV-positive exosomes promote KSHV oral transmission using the 3-D tissue models. We will apply single cell RNAseq to identify cell-specific changes in transcriptome of the oral mucosal and tonsil 3-D tissue models following KSHV infection in response to HIV+ exosomes; and 3) elucidate the role of EGFR-dependent mitogen-activated protein kinase activation in enhanced KSHV transmission by HIV-positive exosomes. Success of the proposed research will advance our understanding of KSHV oral transmission at the molecular level and the underlying mechanisms for developing potential novel therapeutic strategies.