University Of Illinois At Chicago

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Despite the rapid emergence of biophysical tools to detect and characterize conformational changes in protein structure, studying protein dynamics with high sensitivity and reliability in its native environment remains a formidable challenge. Laborious sample preparation and requirement for special equipment present a major obstacle for democratizing these tools. Thus, a simple yet robust platform for characterizing dynamic changes in protein conformation is highly demanded. Using azide-containing hypervalent iodine reagents, we have developed a novel chemoproteomic platform termed Protein Surface Azidation Mass Spectrometry (ProSurA-MS) that detects conformational changes in proteins with unbiased chemoselectivity. Combined with bioorthogonal chemistry, ProSurA-MS allows proteome-wide, site-specific profiling of protein surfaces with wide coverage and reproducibility. ProSurA-MS effectively mapped conformational changes of purified proteins upon denaturation, protein-small molecule interaction, and protein- protein interaction. Additionally, ProSurA-MS detected structural changes in a zinc-binding protein in whole cell lysate upon zinc depletion and measured proteome-wide azidation in live cells, potentiating the characterization of protein dynamics in complex biological environments. The herein proposed ProSurA-MS studies will enable i) characterization of dynamic changes in protein conformation induced by post-translational modifications in response to oxidative stress and monitoring of the protein dynamics of different genetic variants of a metal transporter (Aim 1), ii) basic understanding of the chemical mechanism behind the ProSurA reaction and development of second generation reagents with greater azidation yield and surface coverage (Aim 2), and iii) establishment of a novel method for the identification of protein-protein interactions based on protein surface azidation in live cells (Aim 3).

NIH Research Projects · FY 2026 · 2022-05

New world hemorrhagic fever arenaviruses (NWAs), such as Junín virus, are rodent-transmitted viruses that cause ~30% mortality when they zoonose into humans. The mechanism by the NWAs induce disease is still not certain, although it likely includes induction of high levels of cytokines by infected sentinel cells of the immune system, leading to endothelia and thrombocyte dysfunction and neurological disease. Survivors of Junín infection develop strong humoral immune responses, suggesting that controlling infection at early times post-infection is critical for virus clearance. Although an effective Junín virus vaccine has decreased disease incidence, sporadic cases of this as well as the other known and novel NWAs for which there are no vaccines or effective therapeutics still occur. It is well-established that the clade B pathogenic NWAs bind to transferrin receptor 1 and other receptors on the cell surface, but the steps leading to their entry from an acidic cellular compartment are not well-determined. We recently performed a siRNA screen with pseudotyped viruses bearing a pathogenic Junín glycoprotein with the goal of finding host genes involved in entry that could serve as therapeutic targets. We found that TRIM2, a member of the tripartite motif family that includes well-known members of the host's intrinsic defense against viral infections, limits NWA endocytosis into cells. By probing the TRIM2 interactome for other host proteins that block NWA infection, we discovered that SIRPA, a cell surface protein that inhibits macrophage phagocytosis of tumor and dead cells and erythrocytes, also decreases infection. Importantly, SIRPA, unlike TRIM2, inhibits infection by various human pathogenic viruses that require trafficking to an acidic compartment, including VSV, Zika virus, LCMV and Ebola and SARS-Cov-2 pseudoviruses. Our data suggest that TRIM2 and SIRPA act at the viral entry/internalization step. These finding suggests that there are common mechanisms that regulate virus endocytosis and phagocytosis. We propose here to further investigate the overlap between virus-mediated endocytosis and phagocytosis in vitro, ex vivo and in vivo in three aims that will 1) investigate the overlap in the NWA entry and phagocytosis pathways; 2) determine where TRIM2/SIRPA inhibition of infection occurs; and 3) use TRIM2, SIRPA and other relevant knockout mice to probe the roles of these proteins in cell-type specific and in vivo infection by replication-competent NWAs. In addition to providing mechanistic insight into the entry of NWAs into cells, these studies have the potential of increasing our understanding as to how host factors limit infection and could lead to new approaches to therapeutic intervention.

NIH Research Projects · FY 2025 · 2022-05

ABSTRACT Despite advances in HIV diagnostics, care and prevention strategies, HIV infection rates among adolescents and young adults in the United States (US) remain stubbornly high. There is an urgent need to describe the epidemiology and trajectories of HIV acquisition in this population and to offer age-appropriate, scalable prevention interventions to those at highest risk of infection in the US. This project will engage and retain sexually-active adolescents and young adults in the US in an innovative longitudinal cohort, enroll them in a dynamic established digital health retention platform (HMP; HealthMPowerment), monitor HIV risk and prevention behaviors and explore the socioecological factors that influence the use of new HIV prevention technologies (UG3 phase), while also allowing targeted testing of novel digital health interventions (UH3 phase). We will also test the efficacy of expanding the core version of HMP (HMP Basic) by adding adherence tools (HMP Enhanced) for those who are on PrEP or ART to improve adherence and persistence. In Aim 1, we will enroll and retain a large (n=3000) cohort of sexually active adolescents and young adults, ages 13-34, who are at risk for HIV using innovative digital recruitment, engagement and retention strategies. Over the course of the study, we will longitudinally characterize the sexual behavior, HIV transmission risk, and PrEP uptake trajectories of youth utilizing epidemiological trajectory analyses to identify the most effective points of intervention (Aim 2). For Aim 3, we will launch a randomized clinical trial to examine the efficacy of HMP Enhanced to improve PrEP adherence among HIV-negative youth (n ≥750) and ART adherence among HIV-positive youth (n ≥150) compared to HMP Basic. Finally, we will maximize the productivity of the cohort by testing new and innovative digital health devices, HIV/STI diagnostics and interventions, informed by the previous aims as well as emerging NIH prevention priorities (Aim 4). Our investigative team has decades of experience with recruitment, prevention and care of at-risk youth and large-scale longitudinal cohort studies. This study will capitalize upon productive existing partnerships and digital health expertise to articulate the drivers of the ongoing HIV epidemic among the most vulnerable populations in the US in order to identify the most effective, expeditious and scalable strategies to address this ongoing public health crisis.

NIH Research Projects · FY 2026 · 2022-05

Abstract In contrast to rodents, little is known about neurogenesis in the human brain. The few studies that examined neurogenesis in the human hippocampus have come to vastly different conclusions. Determining the existence and course of hippocampal neurogenesis in the human brain is critical for the understanding of brain function, cognition, putative preventative and therapeutic approaches for the treatment of cognitive decline, Alzheimer’s disease (AD) and related dementia (ADRD). Our previous studies showed that hippocampal neurogenesis persists throughout the 10th decade of life. New neurons were observed in the brains of participants with no cognitive impairments (NCI), as well as in patients exhibiting mild cognitive impairments (MCI) or AD. Interestingly, the number of new neurons was significantly lower in MCI and AD compared to NCI. On the other hand, the number of early differentiating and mature astrocytes was increased in the AD brain. Importantly, higher numbers of neuroblasts were associated with better cognitive performance in the brains of aging, MCI and AD patients. Intriguingly, levels of neurogenesis in the brains of SuperAgers, individuals in their 80ies who exhibit memory performance comparable to people in their 50ies, were significantly greater, compared to age-matched individuals with age-appropriate cognitive function. Nevertheless, the observations above were made using the same neurogenic proxies used in the rodent brain and the nature of cells in the human brain recognized by these proxies is not clear. Evidently, studies that could not detect neurogenesis in the human brain used the same proxies. Thus, the goal of this project is to test the hypothesis that hippocampal neurogenesis persists in the aged and AD human brain and its level is associated with cognitive function. By providing new evidence for the presence of hippocampal neurogenesis using novel tools that would validate previously used proxies. Experiments in Aim 1 will examine the hypothesis that neural progenitor cells have a lower level of proliferation and preferable differentiation into astrocytes leading to fewer new neurons in MCI and AD, using multiplex RNA scope and neurogenic proxies. Experiments in Aim 2 will determine the spatial organization of hippocampal neurogenesis in NCI, MCI and AD, and examine the hypothesis that autonomous and non-autonomous factors in the DG determine the level of human neurogenesis in the aging and AD brain, using a combination of spatial transcriptomics (pciSeq) and RNAseq. Aim 3 will address whether new neurons play a role in cognitive reserve and resilience to AD. Experiments will examine the association between cognitive performance , hippocampal neurogenesis and AD hallmarks in SuperAgers, age-appropriate cognitive performance, MCI and AD patients. In summary, this project will provide novel crucial information about the presence of neurogenesis and its role in hippocampal function in the human aging and AD brain.

NIH Research Projects · FY 2026 · 2022-05

Obesity and cardiometabolic comorbidities are leading chronic conditions among middle-aged and older adults. Unhealthy lifestyle habits and obesity have continued to worsen, especially among those with multiple chronic conditions. Middle to older aged adults with underlying multimorbid conditions are particularly vulnerable and are the target population for this study. This study capitalizes on our decades-long translational research on the efficacious Diabetes Prevention Program (DPP) and DPP-based Group Lifestyle Balance (GLB) interventions; our extensive experience in using electronic health records (EHR) for patient identification and monitoring; and our partnerships with multisector stakeholders in digital health and wellness solutions. This multisite clinical trial uses a 2-stage sequential randomization design to test the adaptive and nonadaptive augmentation of an EHR-integrated, validated base (GLB video) intervention using problem solving treatment (PST), a proven behavior therapy. Adults in the United States (N=1029), ≥50 years of age with a body mass index ≥27 and ≥1 cardiometabolic conditions, will be randomized at baseline to base intervention or waitlist control. Responders to the base intervention, defined by ≥3% weight loss at 6 weeks, will continue the base intervention; participants with <3% weight loss or missing weight data (i.e., nonresponders) will be re-randomized to continue the base intervention alone or augmented with PST coaching via videoconference. Waitlist participants will be re-randomized after a 12-week control period to receive the base or the augmented intervention, but without tailoring based on early weight loss. The base intervention will use EHR-integrated delivery of the self-directed GLB videos, 1 per week for 12 weeks, followed by digital behavior change and motivational messages. The augmented intervention includes base intervention + PST videoconference coaching. All participants will receive a tablet, wireless weight scale, and wearable activity tracker and will be followed for 52 weeks after baseline randomization. Aim 1 is to demonstrate intervention effects on weight loss, behavior change, and patient-reported outcomes. We hypothesize: (1) the augmented intervention will be more effective than the base intervention both among early nonresponders to the base intervention (adaptive) and among participants in the waitlist group (nonadaptive) at 52 weeks; (2) the adaptive augmented intervention will be more efficacious than the base intervention and more efficacious than the waitlist control group at 12 weeks. Aim 2 is to identify predictors of clinically significant (5%) weight loss for individual patients, using sociodemographic, clinical and behavioral engagement characteristics. The proposed interventions are poised to have immediate and widespread impact on access, reach, delivery, effectiveness, scalability and sustainability. This study, if successful, will point the way toward an inexpensive, scalable intervention that would likely be adopted by insurers.

NIH Research Projects · FY 2026 · 2022-05

Title: Regulation of beige adipocyte maintenance and its impact on metabolic outcomes Project Summary/Abstract A potential therapeutic target to curb the global obesity and diabetes epidemic is thermogenic beige fat within white adipose tissue (WAT). Unlike white adipocyte which stores fat, beige adipocytes are stimulated thermogenic fat cells in WAT that robustly consume lipid and glucose. However, Beige adipocytes quickly disappear upon the removal of stimuli, a process further accelerated by age and obesity. This creates a key challenge to explore beige adipocytes as a sustainable therapy for chronic metabolic diseases. A long-term goal of my laboratory is to understand the development and maintenance of beige adipocytes, and the metabolic outcomes. In our preliminary studies using unique mouse models, we demonstrate that beige adipocytes can be induced and maintained by pharmacological and genetical measures. Further, our preliminary results suggest that these induced beige adipocytes are maintained in the older mice, even without continuous stimulation. These exciting findings, although preliminary in nature, suggest novel measures to use beige adipocyte as potential therapeutics to treat obesity, diabetes, and other metabolic diseases. In this application, we will investigate whether the sustained beige adipocytes can improve the metabolic fitness under both health and disease conditions, thereby achieving anti-obesity and anti-diabetes effects. Furthermore, we will elucidate the critical molecular mechanisms by which beige adipocyte identity is preserved. In addition, we will employ cell- type selective transcriptomic approach to comprehensively characterize the molecular components involved in beige adipocyte maintenance.

NIH Research Projects · FY 2026 · 2022-05

Autophagy is a catabolic cellular recycling process that maintains cellular homeostasis and its dysregulation has been implicated in numerous diseases, including neurodegenerative diseases such as Alzheimer’s disease (AD). AD is an age-related neurodegenerative disease that affects more than 5 million people in the United States. Autophagic and lysosomal defects have been observed in AD, including accumulation of autophagic vesicles and lysosomal intermediates as well as defective lysosomal processing of autophagosome contents. Small- molecule autophagy activators that could overcome these defects could potentially halt disease progression through the restoration of cellular homeostasis and the prevention of neuronal cell damage. Our central hypothesis is that small-molecule autophagy activators will restore autophagic and lysosomal homeostasis and exhibit neuroprotective effects that will prevent disease progression and ameliorate Alzheimer’s disease symptoms in vivo. This hypothesis will be tested through the overall objectives of this proposal to optimize an autophagy activator as an in vivo tool compound and drug lead and to evaluate the efficacy of autophagy modulation for the resolution of AD phenotypes in disease-relevant assays and in vitro neuronal models as well as an in vivo model. Our approach is innovative because we have identified mTOR-independent autophagy activators and will identify and validate their unique targets and mechanisms of action in neuronal models to potentially reveal new targets for AD drug discovery. The aims of this proposal will contribute to the achievement of our long-term goal to develop new therapeutics for unmet needs in neurodegenerative diseases. FDA- approved drugs for AD treat the symptoms of the disease but do not improve the underlying cell damage that leads to disease progression, further highlighting the need for novel neuroprotective therapeutic options.

NIH Research Projects · FY 2025 · 2022-05

Cardiometabolic diseases (CMDs), including diabetes and hypertension, impact two-thirds of people in the United States and are among the leading causes of death worldwide. CMDs lead to devastating consequences for those afflicated by them, such as the increased risk of vascular and heart disease, leading to future lethal complications such as myocardial infarction or stroke. While lifestyle factors and genetic predisposition play a role in CMD development, the potential contributions of environmental exposures has not been well-established. Starr County, Texas, is a community with staggering rages of CMDs, specifically diabetes mellitus. Interestingly, our preliminary data indicate that metal and metalloid exposures may increase the risk for CMDs and CMD-related traits in this population. Therefore, we hypothesize that exposure to metals/metalloids contributes to the observed heightened risk of CMDs in this population. To address this, we will leverage data from a well-established prospective cohort from Starr County, Texas. Utilizing already measured urinary metal/metalloid concentrations, we will evaluate longitudinal cardiovascular and metabolic trajectories of blood pressure, hemoglobin A1c, fasting plasma glucose, and homostatic measures of assessment for individual metal exposures and metal mixtures in 600 individuals over 3 years. Given the substantial individual and societal burden of CMDs, it is vital to identify modifiable risk factors in order to empower action for disease prevention, detection, and treatment. Through the project proposed in this application, valuable insight will be gained into the role of metal/metalloid exposures and CMD risk. Lastly, through this proposed research project and training plan, the applicant for this fellowship will receive comprehensive and complementary mentorship from a team of experts with a long history of collaboration, numerous opportunities for publication and presentation of scientific progress, and an overall outstanding education empowering her to become an independent principal investigator and physician-scientist devoted to improving environmental health.

NIH Research Projects · FY 2025 · 2022-05

SUMMARY mRNA translation, or protein synthesis, is a fundamental cellular process that can be dysregulated in several human diseases. Macrophages are heterogeneous populations that are present in most tissues and adopt tissue specific functions. The role of mRNA translational control in macrophages and in regards to their tissue specific functions is not well understood. The broad goal of the proposed studies is to understand how dysregulation of mRNA translation controls tissue-resident macrophage function during stress. The specific goals of this study are to identify how GCN2 (general control nonderepressible 2)-dependent translational control in macrophages affects macrophage function in RBC production and clearance and to uncover the genes mediating this effect. The GCN2 is a serine/threonine-protein kinase that belongs to a signaling network that coordinates cellular response to nutrient stress through translational regulator eIF2 (Eukaryotic translation initiation factor 2). GCN2 senses amino acid levels and phosphorylates eIF2 in response to amino acid deficiency. p-eIF2 inhibits global mRNA translation but paradoxically stimulates the translation of a subset of key stress-response genes such as ATF4 (Activating Transcription Factor 4). Upregulation of stress-response genes in response to GCN2/eIF2 signaling activates a transcription program that helps the cells to overcome unfavorable conditions or undergo apoptosis. GCN2 function has been previously linked to important physiological and pathological conditions such as memory formation, cancer and inflammatory diseases. However, the role of GCN2 in regulating tissue-resident macrophages and their functions in RBC production and clearance has not been characterized. Our current model suggests that GCN2 controls RBC production and clearance during stress through regulation of mRNA translation in macrophages. Therefore, we propose the following aims to achieve our goals: First we will determine the importance of GCN2 in RBC clearance by macrophages (Aim 1) and define molecular mechanisms through which GCN2 impact this process. Next, we will elucidate how GCN2 controls RBC maturation and production by macrophages (Aim 2). Finally, we will examine how mechanical force sense by macrophage through GCN2 (Aim 3). To achieve these goals we will use transgenic mice lacking GCN2 or carrying phospho-resistant eIF2 in macrophages and state-of-art technology to study mRNA translation at genome-wide level. Our mouse models and in vivo and in vitro experiments will rigorously assess the central role of macrophages in development of GCN2-dependent defects during stress. Our genome-wide approach and in vitro functional analysis of selected targets will discover novel translationally regulated genes downstream of GCN2 that play important roles in macrophage regulation of RBC production during stress.

NIH Research Projects · FY 2026 · 2022-04

Project Summary/Abstract Patients with opioid misuse disproportionately utilize emergency health services and are at increased risk for premature death. The timely and accurate identification of patients with opioid misuse in the Emergency Department (ED) is critical to provide evidence-based interventions to decrease mortality. Challenges to opioid misuse detection in the ED include provider time constraints, inconsistent screening approaches, and patient barriers to self-reporting. Advanced analytic techniques such as machine learning and cluster analyses offer promise in efficiently characterizing and identifying patients with opioid misuse during their ED encounter by leveraging data within the electronic health record (EHR) and the prescription drug monitoring program (PDMP). The role of machine learning approaches utilizing multiple data sources to identify ED patients with opioid misuse has yet to be fully explored. In aim 1, multiple machine learning algorithms using ED encounter data will be developed for the identification of opioid misuse. Models will be systematically assessed for social biases and mitigation strategies implemented to ensure solution-oriented approaches in model performance. In aim 2, the inclusion of longitudinal PDMP data for the identification of ED patients with opioid misuse will be evaluated by building models from both data sources utilizing ensemble stacking methods. Finally, in aim 3, an unsupervised latent class analysis model will be built to identify clinically relevant subphenotypes of ED patients with opioid misuse, describe their characteristics, and determine patient-oriented outcomes. An innovative approach to the detection of ED patients with opioid misuse will be pursued by rigorously testing machine learning models utilizing multiple data sources, conducting social bias assessments prior to clinical deployment, and characterizing latent groups of patients with opioid misuse. The candidate for this Mentored Patient-Oriented Career Development Award (Dr. Neeraj Chhabra) possesses a strong foundation in emergency care, medical toxicology, substance use research, and biostatistics. Through this K23, he will further develop skills in data science to build comprehensive and scalable models spanning multiple data domains for the identification of patients with opioid misuse. The multidisciplinary mentorship team led by his primary mentor (Dr. Niranjan Karnik) and co-mentors (Dr. Majid Afshar, Dr. Harold Pollack, and Dr. Gail D’Onofrio) consists of nationally renowned experts in the fields of substance use research, machine learning, natural language processing, and clinical ethics. Through an integrated program of formal coursework, ethics training, mentorship, and research, Dr. Chhabra will develop the skillset necessary to complete these aims and transition to independent investigation. His proposal takes full advantage of the resources provided by the University of Illinois Chicago. Dr. Chhabra’s long-term goal is to utilize machine learning techniques to focus treatments and resources towards patients with opioid misuse within the ED setting. This K23 award provides the necessary foundation to pursue this goal and will form the basis for future R01 proposals evaluating the clinical impact of these models.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY/ABSTRACT Hematopoietic stem cells (HSCs) possess the ability to replenish a new functional hematopoietic system in recipients following transplantation and are the only curative treatment for many hematopoietic malignancies and disorders. However, the availability of donor human leukocyte antigen (HLA)–matched allogeneic HSCs is often too low for successful transplantation. Haplotype mismatch (or haploidentical) transplantation, where donors are matched at half of the HLA loci, is an attractive alternative but is limited by increased rejection, often requiring high-dose immunosuppression to sustain donor grafts. Current clinical transplantation strategies primarily target the adaptive immune response, however innate immune cells have recently emerged as key actors in the allograft response in various transplantation models, contributing to both the success and/or rejection of the transplant. Thus, better understanding of the innate immune response in transplantation and how to target it, is critical. We have recently discovered a novel function for the HSC niche molecule Vascular Cell Adhesion Molecule-1 (VCAM1) on HSCs, in which VCAM1 acts as a “don’t-eat-me” signal for innate immune phagocytes in the context of haplotype mismatch transplantation. Hence, manipulating VCAM1 expression levels may represent a promising immuno-regulatory approach to control the activity of innate immune cells promoting haplotype mismatch engraftment. In our first and second aims, we propose to use mouse models of transplantation to dissect the cellular and molecular mechanisms by which VCAM1 promotes immune evasion from phagocytes in the context of major histocompatibility complex (MHC)-I presentation. In our third aim, we will assess the impact of engineering VCAM1 expression levels on donor stem cells in promoting engraftment across the histocompatibility barrier. This proposal will provide valuable insights in the role of MHC-I engagement in controlling the function of innate immune cells, and illuminate new molecular targets to improve outcomes in immunologically mismatched stem cell transplantation assays.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY ABSTRACT Cervical cancer mortality is expected to increase by 42% to 442,926 deaths worldwide by 2030. Currently 85% of incident cervical cancers and 87% of cervical cancer deaths occur in low- and middle-income countries (LMIC). Due in large part to the low utilization of cervical cancer screening services, Senegal ranks 17th in the world for cervical cancer incidence. For women ages 40 to 49 in rural regions of the country the cervical cancer screening rate is very low at less than 2%. As a result women often present with mostly preventable late stage cancers. Secondary prevention programs (cervical cancer screening) are critical to effectively achieving global progress toward the elimination of human papillomavirus related cancers. However, considerable context variation across geographic- and health system-levels in LMICs markedly obstructs the community responsiveness of an implemented program. Lessons learned from higher resource countries show that incidence and mortality rates decline and early deaths are prevented with early detection and appropriate follow- up and treatment. Prior efforts and pilot studies in Senegal show that there is slow uptake, poor follow-up, and low treatment rates for women who are screened positive. Patient navigation is a strategy to eliminate barriers to screening, timely diagnosis, and follow-up to improve cancer outcomes in vulnerable populations. In high-income countries, patient navigation has demonstrated considerable effectiveness in its ability to address communication, information, medical system, and emotional barriers to timely care across all phases of the cancer care continuum, including detection, diagnosis, treatment, and post-treatment quality of life. Patient navigation programs enhance access to care, promote self-efficacy, and sustain patient engagement with care, and have been shown to improve cancer outcomes, particularly among marginalized groups, rural populations, and impoverished communities. The goal of this project is to prevent unnecessary deaths due to cervical cancer in Senegal. This mixed methods research responds to identified intrapersonal- and community-level barriers to early cervical cancer screening uptake, follow-up, and treatment among women there. We will apply the Dynamic Adaptation Process to study the adaptation of an evidence-based cervical cancer patient navigation program in urban and rural contexts of Senegal, measure the intervention effectiveness, and evaluate programmatic implementation outcomes. By studying the process of adaptation of a patient navigation program in a low- and middle-income country, we will apply implementation science to a novel context. With a particular focus on how the adaptation responds to cancer-related stigma and women’s autonomy in healthcare decision-making, our project demonstrates additional innovation. The process knowledge generated will further our long-term goal to inform the national cervical cancer prevention and control programs in Senegal and other low- and middle-income countries.

NIH Research Projects · FY 2025 · 2022-04

Project Summary/Abstract Hypertrophic cardiomyopathy (HCM) is a common familial cardiovascular disorder viewed as a genetic disease of the sarcomere, since most mutations occur in genes that encode sarcomere/cytoskeletal proteins. Despite decades of basic and clinical research, there are critical gaps in our knowledge concerning how defective biophysical signals in the myocyte influence the function of other cellular compartments of the heart during the clinical course of this disorder. We have reported that early interventions aimed at normalizing myofilament properties only partially prevent HCM progression. Moreover, removal of the triggering mutation does not always reverse progression. In experiments proposed here, we test the overall hypothesis that critical, but treatable, maladaptive modifications in the vascular/endothelial compartment occur early and in parallel with changes in myofilament properties in the progression of HCM linked to thin filament mutations triggering different biophysical and biochemical signals. Preliminary data strongly support a role for and a need to investigate vascular remodeling and endothelial dysfunction that exacerbate symptomatic HCM. Novel data support our focus on HIPPO/YAP/TAZ signaling with emphasis on protective effects of sphingsine-1-phosphate receptor (S1PR) signaling, which is common to the endothelium (EC) and myocytes (CM). Our aims are as follows: Aim 1. Determine the decline in coronary function, changes in vascular remodeling and mechano- sensing in HCM linked to mutationsTnT-R92Q and Tm-E180G with different signaling in progression to HCM. Aim 2. Establish whether restoration of the endothelial HIPPO pathway is sufficient to impede HCM progression. Evidence provided here for a role of EC HIPPO/YAP/TAZ signaling in HCM progression demands an investigation of the consequences of its regulation, and whether therapeutic interventions modify HIPPO signaling. Aim 3. Evaluate the microenvironmental signals responsible for HIPPO pathway dysregulation and co-translation expression of activated YAP/TAZ protective mediators in HCM. Our approach includes determination of the time course of changes in coronary flow velocity, vascular/endothelial histology, and mechano-sensing through key components of the HIPPO pathway, with changes in cardiac function and the myofilaments Ca2+-response during HCM progression. We will treat mouse models early in HCM progression with S1PR agonists, and small molecule inhibitors to normalize myofilament Ca2+ sensitivity and tension to examine whether they restore EC HIPPO pathway and angiogenic signaling. We will identity EC and CM specific disease signaling networks and determine whether HCM leads to impaired S1P export and paracrine function, by enriching and probing the "functional co-translatome" in the RiboTag reporter mice crossed with HCM mutations. Accomplishing our aims will provide discovery of targets for effective and individualized therapies for HCM.

NIH Research Projects · FY 2026 · 2022-04

ABSTRACT Dementia is a major global health challenge, and Alzheimer’s disease (AD) comprises 70% of dementia cases. Because AD has no effective treatment options, the Lancet Commission recently emphasized the critical need for effective, life-course prevention of AD. Epigenetic age acceleration (EAA), or increased DNA methylation (DNAm)-based age relative to chronological age, was identified as a powerful biomarker of AD-related neurobiological substrates and cognitive function in older adults. Likewise, our preliminary data demonstrated associations between EAA and cognitive function in midlife, a critical epoch in brain health when subclinical pathology first emerges, and dementia prevention may be most effective. Works by us and others have also identified associations between cardiovascular disease (CVD) risk factors and EAA, suggesting that EAA could help to explain the intricate link between early life heart and midlife brain health. Despite these intriguing data, prospective associations between childhood CVD risk factors and EAA in adulthood remain unknown and temporal associations are not established. In addition, there is a paucity of research examining the relationships of EAA with midlife cognitive function decline and AD-related neurobiological substrates. We hypothesize that EAA is associated with cognitive decline and neurobiological substrates in midlife and mediates the associations of early life CVD risk factors with these midlife brain health endpoints. To test this hypothesis, we will leverage the rich resources of the Bogalusa Heart Study (BHS), including life-long measures of CVD risk factors, three repeated measures of genome-wide DNAm in adulthood, and two midlife measures of cognitive function over 11-years follow-up in the full cohort of 1,298 BHS participants (850 whites and 448 African Americans). Furthermore, midlife AD-related neurobiological substrates from 3T magnetic resonance imaging (MRI) and amyloid photon emission tomography (PET) scans are also available in a random subsample of 350 BHS participants. As part of the on-going visit cycle (2020-2024), we propose MRI in another random subsample of 350 BHS participants, amyloid PET scans in another random subsample of 50 participants, along with an additional genome-wide DNAm measure in the full BHS cohort. With these data, we will assess prospective and temporal associations of early life CVD risk factors with EAA (Aim 1); examine the associations of EAA with 11-year changes in cognitive function (Aim 2) and neurobiological substrates in midlife (Aim 3); and analyze the mediating effects of EAA on associations of childhood CVD risk factors with midlife brain health endpoints (Aim 4). The molecular characterization of midlife brain health may have broad implications, ranging from the improvement of risk stratification and sub-phenotyping efforts to the pinpointing of molecular targets for drug development. Identifying CVD risk factor precursors to EAA might suggest optimal strategies to prevent EAA and its brain-related sequelae.

NIH Research Projects · FY 2026 · 2022-04

The NIMH 2020 Strategic Plan identifies Global Mental Health as a cross-cutting research theme integral to the institute’s goals. Within global mental health, there is growing recognition of the gaps in understanding how to intervene on social drivers to substantially reduce the global burden of disease attributable to mental illnesses. This R25 application proposes a research education program on social driver interventions in global mental health. The program, Global Mental Health Research on SoCial Drivers Of MeNtal IllNessEs aCross The Lifespan (gmhCONNECT), is based at the University of Illinois at Chicago and George Washington University with a large U.S. and international faculty of senior global mental health researchers. The mentorship network also includes the people living with mental illness from the Global Mental Health Peer Network. Mentees will also engage with practitioners from the WHO, UNICEF, UNHCR, and major humanitarian organizations. In keeping with the requirements of PAR-20-080, this program targets graduate and health professional students, medical residents, postdoctoral trainees, and early-career faculty who are U.S. citizens and permanent residents who are planning to submit or currently funded through NRSAs, K awards, Fogarty Fellowships, or project grant. Building a career in global mental health research involves mastering complex challenges including working with: the legacies of colonialism, power differentials, local culture, agencies and officials, international and local NGOs, and non-specialist and peer providers. The purpose of this program is to facilitate their success as independent researchers and members of the research community in global mental health. Our program highlights intervention research concerning how social drivers impact mental illness, prevention, and care for populations in low- and middle-income countries (LMICs) and other low-resource settings. The specific aims are: Aim 1: Provide training, primarily through a Summer Institute, which advances the trainees’ research knowledge and skills on the ways in which social drivers impact mental illness, prevention, and care and how social drivers and their impact can be addressed through interventions; Aim 2: Provide one-year of focused intensive mentorship (dyadic and triadic) from a US and LMIC pool of multidisciplinary GMH experts to support the mentees’ research interests and career trajectories. Aim 3: Provide a range of synergistic guided learning opportunities including group mentorship, structured peer mentorship, and engagements with people living with mental illness and practitioners at implementing organizations, which will enable trainees to form their unique mentoring networks; Aim 4: Evaluate the impact of gmhCONNECT on the mentee’s networks, knowledge and productivity, with an emphasis on collaboration when evaluating productivity metrics. Successful implementation of these aims will provide promising early-career scientists with more support for their research, an increased success rate, accelerated time for acquiring NIH funding, and generate more scientifically-rigorous collaborative global mental health research.

NIH Research Projects · FY 2025 · 2022-03

Project Summary/Abstract Dental caries represents a common public health problem. Caries involves bacterial invasion, physicochemical dissolution and proteolysis of the mineral and protein components of teeth. Direct bacterial and byproduct interaction with dental pulp cells and odontoblasts in dentinal caries activates a protective repair process of ‘tertiary’ dentin formation. This process requires the recruitment and differentiation of dental pulp stem cells (DPSCs). Clinical therapies such as pulp capping aim to promote dentin regeneration and sustain pulp vitality and consequently endurance of the natural dentition. However, therapeutic dentin regeneration is elusive. The cellular and molecular mechanisms orchestrating dentin-pulp regeneration following infection are not fully elucidated, especially the role of inflammation on dentinogenesis. We have demonstrated that the complement system, which is an important mediator of inflammation and tissue regeneration, is activated in the caries process. A major role for complement and C5a binding to its receptor C5aR in responses to injury is well established. The C5a receptor- like 2 (C5L2) also participated in inflammatory reactions of several pathological conditions, yet, to date no investigation has explored the role of this enigmatic receptor in tissue regeneration and stem cell biology. Here, we propose a significant role for the complement system and C5L2 in DPSC odontoblastic differentiation and reparative dentin formation. Preliminary studies demonstrate that C5L2 expression by DPSCs is quickly increased during odontogenic differentiation, and this expression is potentiated by the inflammatory cytokine TNFα. Moreover, siRNA silencing of C5L2 expression in DPSCs significantly increases the expression of dentinogenic markers like DMP1 and DSPP during odontogenic differentiation. We provide further evidence that p38 map kinase (p38a) plays a key role in DPSC-mediated dentinogenesis. Here, we explore ways of enhancing this odontoblastic function of DPSCs via a novel C5L2 pathway involving p38a signaling. We will define the role of C5L2 in the odontoblastic differentiation of DPSC and characterize the mechanism of action of C5L2 during dentinogenesis. In vivo dentin formation will be evaluated using the mouse pulp-capping/caries model combined with the C5aR, C5L2 and DSPP/p38a knockout mice. The results obtained from this project will shed new light onto cellular and molecular events that orchestrate the initial steps of dentinogenesis by linking the inflammation to DPSC function through C5L2 and p38a pathways. These studies will provide the basis for future potential therapeutic interventions of dentin-pulp complex regeneration and vital tooth preservation.

NIH Research Projects · FY 2025 · 2022-03

Herpes Simplex Virus-1 (HSV-1), a highly transmissible infection, is common and endemic throughout the world. Similar to other human herpesviruses (HHV), HSV-1 maintains lifelong latency inside host that requires immune evasion through various sophisticated mechanisms. Although HSV-1 is equipped with large repertoire (>80) of protein coding genes, the commonly accepted manifestations of viral gene expression during latency are the accumulation of a noncoding transcript and a set of microRNAs (miR). HSV-1 encoded viral miRNAs (v-miRs) are demonstrated to control expression of both viral and host transcripts and regulate viral tropism, lytic switching, immune subversion, etc. While multiple studies have examined HSV-1 profiles in various cell lines, key biological functions of these v-miRs remain unknown. Therefore, we propose to evaluate: (1) systematic expression dynamics of v-miRs during disease progression and reactivation, (2) comprehensive role in the pathogenesis by perturbing immune cells functions and (3) therapeutic targeting of v-miR by synthetic oligonucleotides to mitigate HSV-1-mediated ocular herpes. Using our established mouse model of ocular herpes, we will compare HSV-1 miRNA profiles in primary and reactivated mice corneal tissues and blood. Identifying positive and negative regulatory v-miRs can yield novel insights into host-virus interaction. The information gained will be used in designing therapeutic v-miR Inhibitors to silence candidate v-miRs functions. Effect of synthetic oligonucleotides (v-miR Inhibitors) targeting candidate v-miRs will be assessed on ocular disease progression in a mice model. V-miR inhibitors (alone or in combination) will be topically delivered and virus release, viral transcript/genome, and disease severity score will be measured. Next, we will evaluate how v-miRs can render host immune system dysfunctional, an integral feature required for HSV-1 persistence. Using v-miR inhibitors, we will assess whether immune infiltration and functions can be restored in vivo. Immune cell subsets will be comprehensively profiled in virus-infected animals treated with v-miR inhibitor using flow cytometry and single cell RNA sequencing. In addition, transcript expression profiles of genes related to antigen processing/presentation pathway, critical for potent antiviral response, will be quantified. This will identify the in vivo mechanisms through which v-miRs can facilitate immune evasion in ocular tissues. Next, we will dissect underlying mechanisms of v-miR-mediated dysregulation of antigen processing/presentation by macrophages and dendritic cells and activation of T cells. V-miR expressing myeloid cells will be assessed for uptake and processing of viral antigens and activation of autologous T helper (CD4+) and T cytotoxic (CD8+) cells. In addition, we will assess the impact of v-miRs on the polarization of CD4+ T cells. The data generated will provide significant information to existing knowledge gaps. Overall, the proposed translational study focuses on identifying the therapeutic and mechanistic aspect of v-miRs in ocular disease pathogenesis through modulation of immune cell responses.

NIH Research Projects · FY 2026 · 2022-03

Infection-associated blindness caused by herpesviruses is a leading cause of vision loss in the United States. Frontline therapies include the use of nucleoside analogs such as acyclovir which inhibit the viral thymidine kinase to restrict viral DNA replication. However, emergence of drug resistance and lack of strong corneal bioavailability have made it an urgent priority to develop alternative therapeutics. We have recently discovered a new mechanism through which herpesviruses, exemplified by herpes simplex virus type-1 (HSV-1), propagate in the corneal epithelium. We have shown that endoplasmic reticulum (ER)-localized host protein cyclic adenosine 3′,5′-monophosphate (cAMP) response element-binding protein 3 (CREB3) is essential to HSV-1 replication. Our findings shift the current understanding that CREB3 is only a cellular homolog of HSV-1 VP16. We showed that it is an important pro-viral factor that can be exploited to generate novel therapeutics against HSV infections. We for the first time showed that its modulation via a chemical chaperone 4- phenylbutyrate sodium (Na-PBA), can alleviate, ER stress, reduce CREB3 expression and inhibit viral replication. PBA is currently approved to treat urea cycle disorder. Our translational results are supported by strong in vivo murine data that suggests antiviral efficacy and topical dosage safety of Na-PBA. Due to the high sodium burden associated with Na-PBA administration, its unpalatability, and inability to penetrate sufficiently through corneal epithelium upon topical administration, we have developed various sodium-free PBA nanoformulations to overcome limitations associated with oral and topical delivery of Na-PBA. The purpose of this R24 application is to generate preclinical data in two animal models that support an IND application for repurposing PBA to treat ocular HSV infection. This will be achieved via 3 well thought, exhaustive specific aims. In the first aim, we will evaluate dose-dependent pharmacokinetics, and safety of orally and topically delivered Na-PBA solution and various sodium-free PBA nanoformulations. Furthermore, we will also determine oral and topical, and oral antiviral efficacy of Na-PBA and sodium-free PBA nanoformulations in murine models of ocular HSV-1 infection. The second aim will use the most effective oral and topical formulation(s) and test their safety, PK, and efficacy in guinea pig and rabbit models of primary and reactivated ocular HSV-1 infection. Finally aim 3, we will investigate the potential of Na-PBA and sodium-free PBA formulations to synergize with existing antiviral therapies to determine their potential as an add-on modality to the existing treatment. The latter is likely and significant since PBA is a rare drug that works via alleviating ER stress and aiding the host cell’s response to viral infection, and thereby reducing the chance for emergence of viral resistance. We have assembled a multidisciplinary team including scientists, clinicians, drug development and translation experts who can help us navigate through requisite FDA guidelines. PBA has the potential to become a safe and efficacious alternative to existing ocular antivirals very quickly.

NIH Research Projects · FY 2026 · 2022-02

Project Summary Apidaecin (Api) and Drosocin (Dro), are proline-rich antimicrobial peptides (PrAMPs) produced by honeybees and fruit flies, respectively, which share a unique mechanism of action. Our previous studies of Api showed that upon entering Gram-negative bacterial cells through the SbmA transporter, Api binds in the exit tunnel of ribosomes that have just released the newly made protein and arrests the ribosomes at stop codons by trapping the associated tRNA and release factor. As such, Api represents the first-ever described specific inhibitor of translation termination. Our subsequent whole-genome studies revealed that arresting terminating ribosomes triggers several downstream events that accentuate the inhibitory action of this PrAMP, including ribosome queuing and readthrough of stop codons. Our preliminary data indicate that Dro, despite its distinct amino acid sequence, inhibits the termination step of translation as well, by a mechanism likely resembling that of Api. Their idiosyncratic mode of binding to the target, the unique mechanism of action, and the triggering of downstream effects harmful for the bacterial cell, make these antibacterial peptides an attractive model for developing novel antibiotics. Furthermore, the biological nature of these PrAMPs opens unique opportunities for their screening and optimization by generating hundreds of thousands of peptide variants directly in bacterial cells. In the current proposal we will use the combined effort of three laboratories with expertise in biochemistry and genomics of ribosomal antibiotics, in peptide chemistry and in structural analysis of ribosome-antibiotic complexes to advance the fundamental understanding of the mechanism of action of Api- and Dro-like translation termination inhibitors and identify derivatives with superior on-target activity and expanded spectrum of antibacterial action. In order to achieve these goals we will test arrays of Api and Dro variants in bacterial cells by the tunable expression of peptide gene libraries, determine high-resolution X-ray crystal structures of ribosome-peptide complexes, and employ rational structure-based design to generate via chemical synthesis peptide variants with superior properties. Specifically: In Aim 1, we will identify Api-derived peptides with improved activity upon ribosomes from Gram-negative and Gram-positive pathogens. In Aim 2, the spectrum of action of Api-like peptides will be expanded by bypassing the necessity for uptake by the SbmA transporter. Finally, in Aim 3, we will analyze the ribosome binding and mechanism of action of Dro-like peptides and use comparative analysis to identify the key features that define the class of antimicrobial peptides that target translation termination. The three Aims are tightly interconnected but completely independent from each other. The reagents and tools that will be generated in the course of the proposed work are aimed to serve as leads for future clinical development. Importantly, the results obtained in the proposed studies will significantly advance the fundamental understanding of the properties and mechanisms of action of PrAMPs and will stimulate the progress of the field of ribosome-targeting antibacterial peptides, which currently is still in its infancy.

NIH Research Projects · FY 2024 · 2022-02

ABSTRACT APOE genotype is a major genetic risk factor for several neurodegenerative disorders. Compared to APOE3, APOE4 is associated with greater cognitive dysfunction in older adults, increases Alzheimer's disease risk and exacerbates progression of vascular dementia, stroke, and traumatic brain injury. Evidence supports a major role of APOE4 in brain endothelial cell (BEC) dysfunction at the blood-brain barrier in all these conditions. BEC dysfunction can lead to neuronal dysfunction through disrupting the complex neuronal homeostatic environment and via entry of proteins and other toxins that can damage neurons directly and via effects on supporting cells. Due to their unique location, BEC are susceptible to signals from the brain and plasma in neurodegenerative disorders, which may be particularly relevant for inflammation. Indeed, APOE4, neuroinflammation, peripheral inflammation and BEC dysfunction are intimately linked to dementia risk/progression. Our novel in vitro data demonstrate that APOE4-BECs have a unique basal phenotype that results in disruption of their barrier function with inflammatory stimuli, which we have also found in vivo. Based on these data our hypothesis is that APOE4-associated BEC dysfunction is a novel therapeutic target for neurodegenerative disorders. Our goals are to develop our in vitro assays (R61 Phase) and conduct screening and target identification (ID) (R33 phase) to identify novel compounds that mitigate inflammation-induced permeability disruption in APOE4-BECs. Our biological rationale is that APOE4 predisposes BECs to inflammation-induced barrier deficits, thereby increasing the risk/progression of adult-onset neurodegenerative disorders. The novelty lies in our isolation protocols and assays to target inflammation- induced increases in paracellular permeability using APOE4-BECs. The clinical relevance is that APOE4 is a risk factor for neurodegenerative disorders for which there are also in vivo models. Therefore, there are pathways for the transition of positive hits targeting APOE4-associated BEC dysfunction from preclinical to clinical studies.

NIH Research Projects · FY 2025 · 2022-02

PROJECT SUMMARY / ABSTRACT Coronavirus disease 2019 (COVID-19) is a devastating systemic inflammatory syndrome caused by the coronavirus SARS-CoV-2 which has resulted in over 500,000 deaths in the US during the past year, with this high rate of mortality being attributed in large part to the development of Acute Lung Injury (ALI) or Acute Respiratory Distress Syndrome (ARDS). SARS-CoV-2 entry into cells requires the direct binding of the SARS-CoV-2 spike (S)-protein to the principal host protease-TMPRSS2 and the angiotensin converting enzyme 2 (ACE-2) receptors which are expressed in multiple host cell types. The development of efficacious vaccines to prevent the spread of SARS-CoV-2 represents a tremendous advance that will help curb the COVID-19 pandemic, however the emergence of variants of concern such as the B1.1.7, P1 and B1.351 which can evade the neutralizing responses of the vaccine-induced humoral immune response underscores the urgent need to develop novel therapeutics to complement the vaccination efforts. Based on our provocative Supporting Data, we have formulated the overarching hypothesis that SARS-CoV-2 induced lung endothelial injury is a requisite element of COVID-19 induced maladaptive inflammatory injury that can be therapeutically targeted. We propose the following specific aims: In Aim 1, we will define the nature and underlying mechanisms of lung endothelial injury underlying COVID-19-induced ALI/ARDS. We will test the hypothesis that the degree of lung endothelial injury is a key determinant of the overall pathogenicity and mortality of multiple SARS- CoV-2 variants. We will establish SARS-CoV-2-induced lung vascular injury and compensatory lung endothelial regeneration using two complementary humanized ACE2 mouse models, EC-specific genetic lineage tracing, genetic stabilization of VE-cadherin and single cell RNA-Sequencing. In Aim 2, we will define the efficacy and optimal temporal windows for two targeted pharmacological therapeutic strategies in preventing and resolving SARS-CoV- 2 induced lung endothelial injury. We will test the hypothesis that an engineered soluble hACE-2 peptide has a higher therapeutic efficacy than the wildtype hACE-2 peptide in reducing lung endothelial injury as well as long-term EC reprogramming by preventing viral entry and dissemination of multiple SARS-CoV-2 variants. We will test the corollary hypothesis and that targeted inhibition of IL1β-signaling using a modified IL-1 Receptor antagonist is protective against the feed-forward inflammatory loop and endothelial injury induced by multiple SARS-CoV2 variants. We will use two hACE-2 mouse models as well as compare distinct routes of delivery (intratracheal versus intravenous) and identify the optimal temporal windows for the therapeutic intervention.

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY / ABSTRACT Non-alcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease with a prevalence of 25% in the general population. NAFLD is strongly associated with type 2 diabetes, where it shows a prevalence up to 80%, and a strong progression to non-alcoholic steatohepatitis (NASH, and advance stage of NAFLD). To date, there are no FDA-approved pharmacological treatments for NAFLD. However, thiazolidinediones (TZD), which are effective anti-diabetogenic drugs, may be used to treat NAFLD/NASH. Specifically, TZD activate peroxisome proliferator-activated receptor gamma (PPARγ) in adipocytes, macrophages, and hepatic stellate cells (HSC), and they should reduce insulin resistance, inflammation, and fibrogenesis, respectively. Despite having some positive effects, TZD are not currently used to treat NASH, and it is possible that their true potential as anti- NASH drugs are indeed reduced by direct negative actions on hepatocyte function. In fact, our preliminary studies show that hepatocyte PPARγ is a relevant factor in the regulation of hepatic gene expression that contributes to the progression of NASH, and reduces the therapeutic effects of TZD in the liver of mice with NASH. In this proposal, we hypothesize that hepatocyte PPARγ is a negative regulator of phosphatidylethanolamine methyltransferase (PEMT) and betaine-homocysteine methyltransferase (BHMT), disrupts methionine metabolism, and promotes NASH. In our Aim 1, we will define the contribution of hepatocyte PEMT and BHMT in the TZD-mediated reversal of NASH. Specifically, we will restore the expression of PEMT or BHMT with adeno-associated viruses in PPARγ-intact mice after the development of NASH and treat these mice with TZD to reverse NASH. Also, hepatocyte PEMT and BHMT will be restored in mice with hepatocyte- specific loss of PPARγ expression without a TZD treatment. This aim will show how PEMT and BHMT improve liver health, and reduce steatosis, inflammation and fibrosis to enhance the therapeutic actions of TZD in the reversal of NASH. In aim 2, we will determine if PPARγ directly disrupts the metabolism of methionine in hepatocytes. Briefly, we will use targeted metabolomics in mouse and human primary hepatocytes or in perfused livers of controls and mice with hepatocyte-specific loss of PPARγ expression that are treated with TZD. These experiments will identify how TZD alters the use of methionine in hepatocytes, and contributes, in a hepatocyte- specific PPARγ-dependent manner, to sustain steatosis, inflammation and fibrosis despite the positive actions of TZD on adipocytes, macrophages, and HSC. Overall, in this proposal we will describe how hepatocyte-specific PPARγ negatively regulates methionine metabolism to promote NAFLD, and to limit the potential of TZD as a therapy for NASH. The outcomes of this project will lead us to develop therapeutic strategies that enhance the use of TZD, and to develop new treatments for NAFLD and the care of NASH patients.

NIH Research Projects · FY 2026 · 2022-01

Project Summary/Abstract Alcohol’s effects on the body are complex, altering behaviors, emotions and physiology. These effects all contribute to the cycle of alcohol use and abuse that leads to alcohol use disorders (AUDs). They also contribute to the development of comorbid psychiatric disorders, like panic disorder (PD), which are common. Comorbidity is associated with worse patient outcomes, yet little is known regarding the pathophysiology regulating comorbidity. Emerging evidence supports dysregulated acid-base homeostasis may be a shared mechanism in AUD and PD. Maintaining physiological homeostasis (e.g. neutral pH) is critical for survival, and threats to homeostasis elicit behavioral, emotional and physiological responses directed toward this goal. Alcohol use induces acidosis which is positively correlated with withdrawal severity. Strong evidence supports dysregulated acid-sensing in PD, but its role in AUDs is not well understood. Our lab recently found a role for a novel microglial acid-sensor T-cell death associated gene 8 (TDAG8) in panic-relevant behavior and physiology. Interestingly, preliminary data show ethanol increases TDAG8-promoter driven GFP expression and neuroinflammation in a TDAG8 dependent manner. Thus, TDAG8 and associated neuroimmune effectors may provide a unique and novel shared mechanism for AUD and PD. The purpose of this K99-R00 proposal is to determine if TDAG8 regulates AUD-associated outcomes (K99 phase) and contributes to development of comorbid AUD-PD (R00 phase). My published and preliminary data support the hypothesis that microglial TDAG8 regulates behaviors (ethanol consumption) and physiological (respiratory/cardiovascular) responses associated with alcohol use (K99), and that ethanol-evoked upregulation of TDAG8 increases susceptibility to develop comorbid AUD-PD (R00 phase). Aim1 (K99 phase) will test the hypothesis that microglial acid-sensor TDAG8 regulates ethanol consumption in the “drinking in the dark” (DID) voluntary binge drinking model. Aim 2 (K99 phase) will test the hypothesis that TDAG8 mediates cardiovascular (blood pressure, heart rate, heart rate variability) and respiratory effects of alcohol use and withdrawal (acute and protracted abstinence). Aim 3 (R00 Phase) will test the hypothesis that alcohol dependence will increase panic-relevant behavioral and physiological responses to CO2 inhalation (PD-relevant homeostatic stress) in a TDAG8-dependent manner. Each aim will also investigate neuroinflammation. These aims will elucidate mechanisms of AUDs and comorbidity with PD. Investigating the intersection of AUD and PD by focusing on shared physiological and neuroimmune mechanisms will further our understanding of each disorder and lead to novel treatments. This K99/R00 proposal will provide training in mouse models of AUDs, methods for quantifying neuroinflammation, and collection/analysis of physiological outcomes. Additionally, it will provide professional development that will facilitate my transition to an independent academic research position. These aims will provide a foundation for a successful independent career investigating pathophysiology of AUD and comorbid disorders like PD.

NIH Research Projects · FY 2026 · 2022-01

ABSTRACT The development of type II diabetes (T2D) is strongly associated with obesity, and both are well-established risk factors for cardiovascular disease. Vascular dysfunction is an early event in developing cardiovascular disease in obese diabetic (OB-T2D) patients. Therefore, we set our long-term goal to define molecular mechanisms of vascular dysfunction and corrective strategies that target these mechanisms, such as physical activity and weight loss. We recently discovered that human adipose tissues release extracellular vesicles (adiposomes) that are efficiently captured by endothelial cells. Adiposomes are known to carry bioactive cargos such as proteins and micro RNAs; however, their lipid content has not been studied, neither their ability to transfer their lipid cargo to endothelial cells. In the current application, we propose investigating the role of adiposomes in communicating the unhealthy milieu, mainly dysregulated lipids, to endothelial cells in OB-T2D subjects. On top of these lipid species that we propose to be carried by adiposomes are glycosphingolipids (GSLs). GSLs originate from ceramide glycosylation, a chemical process that is upregulated in the presence of inflammation and high glucose levels. Our preliminary findings showed that in endothelial cells, GSL-rich adiposomes disturb plasma membrane structure and subsequently induces endothelial dysfunction. Moreover, we found preconditioning endothelial cells with high shear stress (which is an exercise mimetic) protected endothelial cells from the detrimental effects caused by adiposomes. Therefore, our central hypothesis is that adipose tissues in OB-T2D patients release GSL-loaded adiposomes that induce vascular endothelial dysfunction. We propose that exercise and weight loss interventions (bariatric surgery) will restore adipose tissue homeostasis, reduce GSL-loaded adiposomes, and subsequently alleviate vascular risk in OB-T2D patients. We will test our hypotheses by pursuing the following Aims: Aim 1: Investigate the role of GSL-rich adiposomes in the pathogenesis of endothelial dysfunction in OB- T2D adults; Aim 2: Test the effectiveness of exercise training in reducing adiposome-mediated effects on vascular function; and Aim 3: Examine changes in adiposome/caveolae axis following metabolic surgery and their association with vascular function. This study will improve our mechanistic understanding of the biological underpinning of adiposome production, packaging, and role in inducing ED under conditions of obesity and T2D. It will also identify adiposomes and the proposed mechanisms of their interaction with endothelial cells as novel therapeutic targets for improving vascular function in OB-T2D individuals. Once these pathways are elucidated, strategies for targeting them can be advanced, leading to an improved therapeutic management of T2D-related cardiovascular disease.

NIH Research Projects · FY 2024 · 2021-09

SUMMARY Cesarean deliveries (cesareans), when used judiciously, save lives. However, research indicates that the use of cesareans in the United States (US) has risen well above the level of necessity and has become a contributor to maternal morbidity and mortality. Patients who have cesareans face increased risk of serious complications such as amniotic fluid embolism, catastrophic hemorrhage, placental abnormalities in future pregnancies, and death. Babies born by cesarean face increased risk of respiratory complications, and, emerging research suggests, chronic disease. For these reasons, the Healthy People 2020 goal is to reduce cesarean rates to ≤ 23.9% for first-time mothers. Currently, over one-third of all women giving birth in the United States deliver via cesarean. Hospital cesarean rates across the US range from 6% to 69%, and significant variation remains even when controlling for patient clinical factors and hospital and patient demographics. Patient request for a cesarean make up less than 1% of cesareans nationwide, and, thus, cannot explain this variation. Our prior work has found that this variation in practice is associated with labor and delivery unit culture and, moreover, that successful reduction in cesarean overuse correlates with specific aspects of unit culture, such as decreased fear of vaginal birth, physician acceptance of oversight, agreement with evidence-based practices, and belief in the importance of maternal agency. Organizational culture has been shown to be a key element in reducing variation in hospital outcomes in other clinical areas; and, underscoring its importance, the national safety bundle addressing cesarean overuse charges hospitals to “develop a unit culture that supports vaginal birth.” Unfortunately, there is evidence that this aspect of quality improvement receives the least attention from hospitals that participate in initiatives to reduce cesarean overuse. After 2 years of a California statewide initiative, about 42% of hospitals still had cesarean rates over the Healthy People 2020 goal. This is not surprising given that few tools and no rigorously designed user- centered tools exist to help hospitals implement culture change. This proposal leverages the strength and support of multiple state-level perinatal quality collaboratives at different stages of implementation and a novel, continually improving Labor Culture Survey tool. We will engage in a comprehensive user-centered design process to create, adapt, and implement a “Culture Change Toolset” for use in hospital-based, organizational culture quality improvement efforts to reduce cesarean overuse. Furthermore, we will incorporate patient perspectives and feedback as key stakeholders in the design and adaptation process. Based on our prior data, a 1-point improvement in unit culture supportive of vaginal birth would convert 331,773 cesarean deliveries or 27% of US cesarean births and 9% of all US births each year to vaginal deliveries and the low-risk, primary cesarean rate would decrease from 28% to 19%, thus achieving the Healthy People 2020 goal and improving maternal and newborn morbidity and mortality.