University Of Illinois At Chicago

NIH Research Projects · FY 2026 · 2024-02

ABSTRACT In the United States, severe obesity (BMI ≥ 40.0 kg/m2) disproportionally affects non-Hispanic Black females of reproductive age. Severe obesity is a significant predictor of adverse perinatal outcomes including gestational diabetes mellitus (GDM), pre-eclampsia, premature birth, and at its most severe, fetal death, birth defects and a three-fold greater risk of maternal mortality – outcomes that also disproportionately affect Black females. Observational studies suggest weight maintenance and even modest body fat loss and altering the maternal metabolic milieu (availability of glucose and lipids) in the gestational period may be important to reducing perinatal health risks among pregnant females with severe obesity. Existing lifestyle interventions (calorie control/dietary pattern changes/physical activity) have showed modest effects on attenuating excess gestational weight gain and modest effects on perinatal health outcomes among pregnant females with obesity. Time- restricted eating (TRE), where an individual simply watches the clock and consumes calories within a particular eating window, is a simple and highly accessible eating pattern that has the potential to minimize gestational weight gain, reduce excess glucose and lipids, and improve metabolic health among pregnant females with severe obesity all of which could translate to improved perinatal health outcomes. Yet there are no clinical trials of TRE in pregnancy. In response to PAS-20-160 - Small R01s for Clinical Trials Targeting Diseases within the Mission of NIDDK, we aim to test the safety, feasibility, and acceptability of TRE among 60 pregnant Black females with severe obesity. We will examine the preliminary efficacy of TRE (8-hr eating window in the 2nd trimester & 10-hr eating window in the 3rd trimester) vs. Standard Care on safety, feasibility, and preliminary efficacy on weight, cardiometabolic risk markers and perinatal health outcomes. The intervention will begin at ~16 weeks gestational age (GA) and continue through admission for labor and delivery. The study will involve weekly meetings with a research team member, and research visits at baseline (~16 weeks GA), 25-27 weeks GA and 35-37 weeks GA. Maternal and fetal health monitoring will occur weekly through a combination of research specific assessments and clinical data collected during routine prenatal care and at labor and delivery via patient electronic health records. The goal of our work is to reduce and prevent adverse perinatal outcomes among pregnant Black females including those with severe obesity. In alignment with NIDDK’s mission, this small-scale R01 study will provide the necessary data to accelerate the initiation of a full-scale efficacy trial of TRE in pregnancy. Given the relative simplicity of TRE it can be easily disseminated in clinic demonstrating its strong potential for wide-scale public health impact.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Cholesterol is a central molecule in human health, serving as a precursor for active metabolites, a specific ligand for proteins, and a determinant of membrane architecture. Dysregulated cholesterol metabolism has wide- reaching consequences and is a hallmark of nearly every major human disease, including neurodegeneration, cancer, and immune disorders. Despite the immense roles of cholesterol and its metabolites in human health, their molecular signaling mechanisms remain largely unknown. The main barrier to understanding and targeting cholesterol metabolites in therapy is our lack of tools to define their precise interactions. To address this gap, we have established innovative cholesterol metabolite probes that contain functionalities for chemoproteomics- based target identification and microscopy-based subcellular localization. We have used these probes to quantitatively profile the proteome-wide interactomes of oxysterol metabolites and specify their roles in specific processes of cancer and immunity. Importantly, our tools have uncovered highly selective protein interactors, including the tumorigenic and neuroactive sigma-2 orphan receptor (TMEM97), providing new opportunities to target cholesterol metabolite activities in disease. Over the next 5 years, we will build upon our established tools and technologies to interrogate oncogenic, immunomodulatory, and neuroactive cholesterol metabolites as well as cholesterol itself. Specifically, we will design a next-generation chemoproteomics platform for multiplexed cholesterol metabolite target profiling and pioneer an integrated fluorescence microscopy approach for subcellular cholesterol metabolite imaging. Our tools and technologies will revolutionize the analysis of cholesterol metabolite interactions, advancing cholesterol metabolism as a druggable target in human disease.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Diabetic retinopathy (DR) is the leading cause of preventable vision loss in the United States and disproportionately affects individuals with limited socioeconomic resources. Timely diagnosis and treatment can prevent up to 90% of vision impairment, underscoring the importance of consistent adherence to ophthalmology appointments. Dr. Scanzera’s prior work identified transportation challenges as a key factor contributing to missed appointments among patients coming from neighborhoods with limited socioeconomic resources. Her K23 research plan aims to design, implement, and evaluate a ride-share transportation intervention adapted from her health system’s existing PROgram for Non-emergency TranspOrtation (PRONTO). This intervention, names PRONTO-EYE, is intended to reduce transportation challenges and improve adherence to scheduled ophthalmology visits for patients with DR and Medicaid insurance. To achieve this, Dr. Scanzera will apply implementation and human-centered design methodologies to (1) co-design and (2) pilot test PRONTO-EYE. She hypothesizes that the intervention will be successfully integrated into the existing healthcare infrastructure and will enhance appointment adherence in the target population. Dr. Scanzera’s long-term goal is to develop and implement evidence-based strategies to reduce preventable blindness in communities with limited resources across the U.S. The K23 Career Development Award will provide her with mentored research experience, as well as didactic and hands-on training, to support her transition to an independent investigator focused on improving eye health outcomes for all. She will benefit from the extensive resources and mentorship available at the University of Illinois Chicago, with advisors whose expertise aligns with her research and career objectives. This proposal also aligned with the National Eye Institute’s 2021 Strategic Plan which emphasizes population health. By developing a transportation intervention tailored to patients with DR with Medicaid insurance, Dr. Scanzera’s research has the potential to significantly improve access to eye care and reduce vision loss. The immediate outcome will be a pilot-tested intervention, which will be evaluated in a larger trial as part of a future R01 grant.

NIH Research Projects · FY 2026 · 2024-01

This study examines differences in reactive stepping responses between older adults with mild cognitive impairment and cognitively intact older adults when exposed to novel and repeated gait-trip perturbations. Falls occur in >30% of older adults each year, most often due to trips, and lead to ~2.8 million hospital admissions, 32,000 deaths, and $50 billion in medical costs. Fall incidence is doubled in older adults with mild cognitive impairment (OAwMCI), which is a prodromal stage of dementia that affects about 17% of the aging population. Substantial evidence indicates that OAwMCI have impaired volitional balance control and gait compared to cognitively intact older adults (CIOA); however, the underlying mechanisms that explain why OAwMCI have a greater predisposition to falls are yet to be investigated. The main causative factors of falling have been identified as deficits in reactive balance control, including reduced center of mass (COM) stability and insufficient vertical limb support post-perturbation, which could be associated with impaired neuromuscular coordination (reduced muscle synergies). Further, evidence from mobile neuroimaging techniques such as electroencephalography (EEG) suggest the role of the cortex in generating a reactive stepping response, as indicated by perturbation- evoked potentials (e.g., N1) and increase in beta frequency power when exposed to unpredicted perturbations. However, it is unknown how the pathology related to MCI (structural neural damage, reduced sensorimotor integration) affects reactive balance responses. Further, it is unknown if OAwMCI are capable of improving their reactive balance through repeated perturbations. Perturbation training has been introduced as a novel fall prevention paradigm involving repeated exposure to simulated balance disturbances (e.g., slips, trips), with robust effects on laboratory and real-life fall reduction. Trial-by-trial analyses indicate that CIOA can acquire motor adaptations in as little as 5 perturbation exposures, including improvements in biomechanical outcomes (fall rate, COM stability) and enhanced neuromuscular control (increased muscle synergy number). In theory, perturbation training could be implemented in OAwMCI to mitigate increased fall risk, however it is first necessary to assess 1) the magnitude of reactive balance impairment, and 2) the ability to acquire motor adaptations to repeated perturbations in this population. Thus, we propose a mechanistic investigation to determine differences in the biomechanical, neuromuscular, and cortical control of reactive balance between OAwMCI and CIOA when exposed to a novel treadmill gait-trip (Aims 1&2). We will also examine differences in the rate and magnitude of motor adaptation between OAwMCI and CIOA when exposed to 8 repeated treadmill gait-trips (Aim 3). The clinical impact of this study will be to mechanistically understand how pathology related to MCI may contribute to balance impairment and increased fall-risk. Further, with an understanding of how OAwMCI are able to adapt to repeated perturbations, this study will pave the road for the future design of effective perturbation training protocols that could significantly improve balance control and reduce fall incidence in OAwMCI.

NIH Research Projects · FY 2026 · 2024-01

ABSTRACT Adverse reproductive outcomes, including preeclampsia and hypertension, preterm birth, growth restriction, and small for gestational age birth contribute significantly to maternal and infant morbidity and mortality and can lead to long term health consequences for both mother and offspring. The placenta plays a central role in successful pregnancy outcomes, fetal development, and it is increasingly being recognized for playing an important role in developmental programming of the fetus and contributing to childhood health and disease. Environmental chemicals are known to contribute to the risk for adverse reproductive outcomes and these chemicals may be eliciting their toxic effects through impacts on the function of the placenta. One particular function of the placenta that is only beginning to be fully understood is its role as a peripheral molecular clock. Disruptions to the molecular clock are recognized to be involved in several adverse reproductive outcomes and recent data suggests that deregulation of molecular clock genes impacts important placental cellular processes such as cell migration. There is also evidence to suggest that the epidermal growth factor (EGF) maybe a central regulator of the placental molecular clock and that a mixture of toxic chemicals, including pesticides, alters the placental molecular clock through inhibition of EGF signaling through the EGF receptor (EGFR). These findings support the premise that a properly functioning molecular clock is required for placental health. However, it remains unknown if environmental chemicals cause placental dysfunction via molecular clock disruption. This consortium, consisting of a placental toxicologist (Veiga-Lopez), circadian biologist (Hoffmann), and pregnancy molecular epidemiologist (Marsit), has developed a project that will lead to an understanding of how gestational environmental exposures impact the molecular clock resulting in placental dysfunction, by examining the overall hypothesis that chemicals that interfere with molecular clock signaling through EGFR lead to placental cell dysfunction in a gestational age- and placenta layer-specific manner. In a mouse model, we will examine how gestational age, placenta layer, and sex impacts molecular clock sensitivity to chemical exposures. In human placenta cellular models, we will identify mechanisms through which the molecular clock proteins PER1, PER2, and PER3 impact cellular function upon chemical exposures. Finally, using a pregnancy cohort of farm-working women and their offspring, we will determine the association between pesticide exposure and placental molecular clock gene dysregulation and build causal evidence by comparing results to those from the mouse model. Through this new and exciting collaboration, there is a unique opportunity to comprehensively demonstrate the importance of the placenta as a peripheral clock and to provide evidence that exposures relevant to the human population impact the placental molecular clock which has lifelong implications for health and disease.

NIH Research Projects · FY 2026 · 2024-01

ABSTRACT More than 2 billion people are projected to be overweight and 1 billion to be obese by 2030. In addition to its specific co-morbidities, obesity is a major risk factor for metabolic diseases and cancer. Emerging approaches utilizing brown adipose tissue (BAT) activation and thermogenesis have the potential to be safe, effective, sustainable, and affordable for treating obesity-related metabolic diseases. However, currently imaging methods for BAT metabolic function have limitations. Recent studies found, through the futile creatine cycling, creatine promotes adrenergic activation of BAT and thermogenesis in a manner that is independent of mitochondrial uncoupling protein 1 (UCP1). We have made breakthroughs in extracting the molecular Chemical exchange saturation transfer (CEST) contrasts from high-resolution Z-spectral imaging of adipose tissues, particularly the creatine CEST (CrCEST) as an endogenous and clinically translatable MRI. Our preliminary animal studies demonstrated that dynamic CrCEST MRI detected significant BAT adrenergic activation in response to a β3-AR agonist CL-316243 (CL). Our central hypothesis is that endogenous CrCEST MRI measures metabolic function of adipose tissues through adipocyte creatine, the mediator of adrenergic activation. Aim 1. To investigate the molecular origin of CrCEST MRI for BAT metabolic function, including characterizing the percentage contribution of creatine to CrCEST and the longitudinal repeatability with and without respiratory gating. Aim 2. To validate CrCEST MRI for the assessment of BAT metabolism in genetically modified mice. We will develop fast dynamic CrCEST MRI based on key-hole technique. We will assess the metabolic balance of UCP1-KO mice through futile creatine cycling. Mice will receive daily saline, Cr supplement, or β-GPA (a Cr kinase inhibitor) for 2 weeks, while dynamic CrCEST MRI and 18F-FDG PET/CT will be performed with biopathological measurements in the end for validation. We expect, Cr supplement will boost the adrenergic activation of BAT in UCP1-KO mice while β-GPA blunt the adrenergic activation. Similarly, we will study adipose-specific Cr transporter knockout (CrTKO) mice and expect that CrTKO impairs adrenergic activation in BAT and disable the effect of Cr supplement. Aim 3. To evaluate longitudinal Z-spectral MRI for monitoring BAT activation and WAT beiging during the prevention or treatment of obesity. In the prevention study, young mice on high-fat diet (HFD) will receive saline, Cr supplement, mirabegron (a β3-AR agonist), or the combination in the diet for 8 weeks and be monitored every two weeks with Z-spectral MRI producing CrCEST and amide proton transfer effect. In the treatment study, obese mice will be treated similarly and monitored for 8 weeks followed with biopathological measurements. The successful development and validation of CrCEST MRI in adipose tissues will provide 1) real-time non-invasive functional imaging of adipose tissues; 2) a better understanding of creatine in BAT adrenergic activation and white-fat browning, and its potential for preventing and treating metabolic diseases.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT While rodents have provided valuable information on key aspects of the gut-liver axis, there are fundamental species-specific differences in the gut-liver axis, such as the composition of the microbiome, differences in liver xenobiotic metabolism, and progression of alcoholic and metabolic fatty liver disease. Therefore, human- relevant in vitro models now play important roles in complementing in vivo animal studies for basic science and for drug screening. While considerable progress has been made in the development of human models of intestine and the liver using physiologically relevant primary cells, very little has been done in engineering an integrated human gut-liver model using primary cells. We have utilized droplet microfluidics to fabricate highly monodisperse extracellular matrix (ECM)-based 3D liver microtissues containing primary human hepatocytes and liver sinusoidal endothelial cells that are rapidly generated and functionally outperform self-assembled hepatic spheroids and hepatocytes within bulk gels for 4+ weeks; furthermore, the microtissues can be further augmented with human hepatic stellate cells and Kupffer cells. We have also separately developed in vitro human intestine models using primary cells, which can be engineered to maintain gradients of O2 across the basal and luminal surfaces to recapitulate the stem cell and differentiated cell compartments of the crypt; can produce mucus; and are amenable to the incorporation of microbiota in the luminal compartment. Here, we will engineer in vitro human gut-liver models of increasing complexities, which will be used to elucidate how intestine, liver, and microbiota interact to modulate reciprocal functions. In aim 1, we will elucidate reciprocal functional changes in a human gut-liver model containing primary human colonocytes and liver cells, while in aim 2 we will introduce mucus and microbiota in the intestinal model and elucidate reciprocal crosstalk upon co-culture with human liver microtissues. This proposal will yield a first-of-its-kind scalable human gut-liver model containing primary cells for in vitro applications, such as compound screening and disease modeling.

NIH Research Projects · FY 2026 · 2024-01

SUMMARY Protein synthesis is an essential process in all living organisms catalyzed by the ribosome – one of the largest and the most complex macromolecular machines created by nature. Our group uses structural, biochemical, and microbiological techniques to gain insights into the molecular mechanisms of ribosome functioning in bacteria, the modes of action of ribosome-targeting antibiotics, and mechanisms of drug resistance. Interaction of the growing polypeptide chain with the nascent peptide exit tunnel (NPET) in the large ribosomal subunit during protein synthesis plays a fundamental role in the regulation of gene expression as well as in co-translational protein folding. Despite the extreme chemical diversity of its substrates and products, ribosome efficiently makes all cellular proteins. However, some of the peptide sequences were evolutionarily selected to be problematic for the ribosome resulting in translation arrest during their synthesis. The cell often uses this phenomenon of peptide sequence-specific translation arrest, which can occur in a ligand-dependent manner turning the ribosome into a small-molecule sensor, to control its metabolism. Currently, we lack a mechanistic understanding of how interactions of the arrest peptides with NPET and small molecules or antibiotics result in the arrest of translation. This knowledge gap could be addressed by structural studies capturing both arrested and non-arrested Ribosome Nascent chain Complexes (RNCs) containing relevant peptides in the NPET. While many cryo-EM structures of ribosomes carrying various stalling peptides became available in the past years, they all represent arrested RNCs. Because the experimental approach used in these studies for the preparation of RNCs relies on the ability of the ribosome to stall in the presence of a small molecule and a wild-type arrest peptide, using the same methods for the preparation of respective non-arrested RNCs is impossible. Alternatively, the desired arrested or non-arrested RNCs could be assembled from the individually purified components. However, this strategy has remained challenging due to the lack of a reliable method for the large-scale preparation of peptidyl- tRNAs. Recently, we have developed a chemoenzymatic approach based on native chemical ligation reaction for facile semi-synthesis of peptidyl-tRNAs. Most importantly, by determining the first high-resolution structures of non-arrested RNCs, we found that synthetic peptidyl-tRNAs can be efficiently complexed to the ribosome in vitro and yet represent a functionally significant state of the ribosome’s catalytic site. Therefore, our group is best suited to bridge the existing knowledge gap in our understanding of molecular mechanisms underlying antibiotic- or sequence-dependent ribosome stalling by determining high-resolution structures of model RNCs under arresting or non-arresting conditions. Altogether, the proposed project will uncover enigmatic mechanisms of sequence-dependent nascent chain-mediated ribosome stalling, provide the structural basis for the context- specific action of several chemically unrelated classes of ribosomal antibiotics, and reveal mechanisms of action of ABCF ribosome protection proteins upon stalled ribosomes.

NIH Research Projects · FY 2026 · 2023-12

Project Summary Chronic overconsumption of high fructose-containing food and beverages has emerged as a significant risk factor for development of obesity, nonalcoholic fatty liver disease, and type 2 diabetes mellitus. Fructose metabolism has been extensively investigated in the hepatocyte, where 50-70% of dietary fructose is metabolized as well as in the small intestine and kidneys, where the fructose transporter Glut5 is highly expressed. However, much less is known in other metabolic organs including skeletal muscle, brain and white adipose tissue. Thus far, the impact of Glut5 and fructose metabolism in intrascapular brown adipose tissue (iBAT) has not been investigated. While analyzing the diet/feeding-stimulated iBAT transcriptomes to gain mechanistic insights, we observed a dramatic but transient upregulation of facilitated glucose/fructose transporter type 5 (Glut5, [Slc2a5]) in iBAT as early as 1 h after feeding from both male and female C57BL/6J mice but not in antibiotic-treated mice. Consistently, we also observed similar Glut5 upregulation in differentiated human brown adipocytes after incubation with serum from fed mice. In addition, we also observed that chronic high fructose consumption dysregulates iBAT basal Glut5 expression. This is unexpected because Glut5 expression in white fat is fructose-independent. Our data showed for the first time that iBAT Glut5 regulates circulating fructose homeostasis and mediates fructose-dependent iBAT whitening and obesity. Fructose metabolism is important for glycogen and lipid accumulation in hepatocyte. Increase in iBAT fructose metabolism due to fructose-induced Glut5 dysregulation could potentially leads to iBAT whitening. Together with our preliminary findings that feeding-induced Glut5 expression in iBAT is eliminated in antibiotic-treated mice, we hypothesize that high-fructose consumption promotes whitening and impairs iBAT metabolic activity via microbiome-dependent iBAT Glut5 expression. In this proposal, we plan to delineate how brown adipocyte Glut5 impacts iBAT metabolism. We will also determine whether high-fructose diet increase Glut5 via microbiota-derived hyodeoxycholic acid (HDCA) to inhibit iBAT function. Finally, we will determine liver-x-receptors (LXRs) expressed in brown adipocyte are required for whitening and metabolic activity impairment in iBAT induced by high-fructose diet. Successful completion of this proposed study will contribute to our understanding of high fructose diet-induced brown fat whitening and metabolic dysfunction.

NIH Research Projects · FY 2026 · 2023-12

The Association for Research in Otolaryngology (ARO) is requesting continued support from an NIDCD Conference Grant for its annual Midwinter Research Meeting (MWM). The MWM is a unique meeting where current basic and clinical research related to otolaryngology is presented. In addition to poster presentations and podium sessions where cutting edge research is presented, the meeting includes symposia and workshops that allow presentation of up-to-date summaries of broad scientific issues that extend beyond the field of otolaryngology. These symposia typically include invited scientists and clinician-scientists from related fields whose work may be relevant to emerging areas of research in otolaryngology. The symposia have been supported by the Conference Grant for more than 25 years and have contributed to the success of the meeting, as evidence by its growth over the years. The growth and development of otolaryngology research depend on bringing young investigators into the field. Travel awards for young investigators, including residents, medical students, and pre-doctoral and postdoctoral fellows have been supported by the NIDCD Conference Grant. We request support to continue these activities. Modified

NIH Research Projects · FY 2026 · 2023-11

PROJECT SUMMARY Polymicrobial infections are a significant health concern to humans. Multi-species biofilms exhibit greatly increased antibiotic resistance, and systemic infections with more than one etiologic agent are difficult to treat and often more lethal than infections with a single pathogen. Salmonella enterica serovar Typhimurium (STm) is a food-borne bacterial pathogen that causes gastroenteritis in over 1.35 million people in the US every year. Recently, a clinical study with 2500 patients uncovered that the under-studied fungal component of the microbiome is an important modulator of Salmonella enterica infections. When patients were colonized with the opportunistic pathogenic yeast Candida spp, they were more likely to be infected with S. Typhi or S. Paratyphi. C. albicans is a frequent gut colonizer and can be found more than 60% of people. The commensal yeast can easily transition to become pathogenic and breach epithelial barriers with its filamentous hyphae. It is also associated with gut inflammatory diseases like Crohn’s disease. The two gut pathogens STm and C. albicans thrive in an inflamed intestine and are likely to co-occur frequently. Nevertheless, the role of C. albicans during Salmonella infection is currently completely unknown. Our long-term goal is to understand the role of gastrointestinal fungi during STm pathogenesis. The specific goal of this proposal is to elucidate the mechanism of how the presence of C. albicans in the gut increase STm colonization and systemic dissemination. Our data show that commensal C. albicans abundance in the mycobiota increased after STm infection of mice and positively correlated with disease severity. During co-infection with STm and C. albicans, mice showed significantly increased STm load in cecum, spleen and liver, and more weight loss compared to STm single infection. Despite two pathogens present, the early host immune response to co-infection was significantly blunted compared to single infections. The lower inflammatory response might result in inefficient clearance and increased dissemination of STm. In the presence of C. albicans in vitro, STm upregulated virulence genes and showed significantly increased invasion of colonic epithelial cells. We hypothesize that the increased virulence of STm in the presence of C. albicans is the result of two distinct mechanisms: (A) direct bacteria- fungi and (B) host-dependent interactions. With this proposal, we will therefore determine in Aim 1 which C. albicans factors directly modulate STm virulence gene expression, mediate binding to STm, and result in higher STm dissemination in two mouse models. In Aim 2 we will elucidate which host cells, signaling molecules and microbial determinants are involved in modulation of the immune response during co-infection. In summary, we will study the cross-kingdom interaction of two important gut pathogens and will provide mechanistic detail on how the presence of C. albicans modulates STm pathogenicity.

NIH Research Projects · FY 2025 · 2023-09

Abstract Converging human association studies link cannabis abuse during adolescence with an increased likelihood of developing cognitive and emotional regulation deficits later in life, many of which are refined and dependent on prefrontal cortex (PFC) maturation during adolescence. However, the neurobiology underlying this adolescent vulnerability remains unclear due to our limited knowledge on how adolescent cannabis exposure impacts the maturation of neural circuits. Our long-term goal is to identify sensitive neurodevelopmental processes that are vulnerable to repeated cannabis exposure, with emphasis on PFC neuronal circuits that undergo functional remodeling during adolescence. Data from our recent studies revealed that a hallmark of PFC maturation is the re-calibration of an excitatory-inhibitory (E-I) balance during adolescence that is required for sustaining proper PFC-mediated behaviors in adulthood. Our data also indicate that it is the maturation of GABA function that renders the PFC labile during adolescence, a developmental process that is intimately linked to the activity of afferent transmission from the ventral hippocampus. Similarly, adolescent, but not adult, exposure to THC impaired the maturation of PFC GABA function. This raises the exciting possibility that the PFC deficit resulting from adolescent THC exposure is mechanistically linked to the disruption of specific inputs driving PFC maturation. Based on our preliminary data, we will test the central hypothesis that the developing PFC GABA circuit during adolescence is highly sensitive to the negative impact of cannabis through a ventral hippocampal -mediated mechanism. Thus, the rationale for undertaking this research is that if PFC GABA maturation is compromised by adolescent THC exposure, the normal facilitation of PFC inhibitory control will be arrested, which in turn could lead to an enduring state of PFC disinhibition resulting in behavioral deficits in adulthood. We will fill this gap in knowledge through the pursuit of 3 Specific Aims. We will use a recently developed combustion/smoking chamber to deliver THC at 3 non-overlapping adolescent windows to establish the precise period during which changes in PFC GABA function (Aim 1) and PFC-sensitive behaviors (Aim 2) are susceptible to repeated THC exposure. Currently available input-specific DREADD manipulations will be used in Aim 3 to establish whether the enduring PFC GABA deficit elicited by THC is causally linked to disruption of ventral hippocampal-to-PFC transmission.

NIH Research Projects · FY 2025 · 2023-09

Employment arrangements across industries and demographic groups have become increasingly precarious in recent decades. This shift has resulted in a growing proportion of the workforce that is exposed to risks and vulnerabilities associated with insecure and substandard employment. There is an urgent need to understand the role of employment precarity in increasing an individual's risk of adverse behavioral health outcomes, which are particularly prevalent among working-age adults and result in considerable loss of productivity in US workplaces. To address this critical issue, robust analyses of existing longitudinal data which allow for comprehensive measurement of individuals' experiences with employment precarity and their relationships with behavioral health are needed. Few existing studies, especially in the US, have sought to measure employment precarity comprehensively and longitudinally and, to date, no such studies have explored the relationships between the precarity of individuals' employment trajectories and their behavioral health. Existing studies of employment precarity have also failed to account for interruptions in employment and changes in jobs over time, despite the known impact of unemployment on adverse behavioral health outcomes. In light of these gaps in existing research, the proposed study will employ data from two National Longitudinal Survey of Youth (NLSY) cohorts (1979 & 1997) to define measures of employment precarity and employment variability and characterize their longitudinal trajectories (Aim 1) and model associations between employment precarity and substance use, two indicators of behavioral health (Aims 2 and 3). The extensive nature of the NLSY79 and 97 datasets will allow for characterization of disparities in employment precarity trajectories and cumulative employment variability, with the potential to elucidate key risk factors affecting the prevalence of precarious work. This pursuit, coupled with the proposed test of whether trajectories of employment precarity and variability are associated with adverse behavioral health in these two longitudinal datasets, addresses research gaps highlighted in the 2020 NORA Healthy Work Design and Well-Being report and can inform opportunities for intervention addressing intersections of work, psychological distress, and substance misuse across the life course. This study provides an excellent opportunity for mentored training and career development focused on the expansion and refinement of skills and knowledge necessary to purse the proposed research aims and career goals. My proposed career development plan will 1) build my understanding of behavioral health outcomes and mechanisms associated with employment, 2) expand my knowledge of advanced survey methodology, 3) develop my competency in statistical concepts for longitudinal analysis of survey data, and 4) strengthen my practical skills in statistical computing for the analysis of longitudinal survey data. These activities, in conjunction with the proposed research strategy, will facilitate my development as an independent researcher poised to be highly competitive for external funding and to significantly impact the field.

NIH Research Projects · FY 2026 · 2023-09

Ocular inflammatory diseases are the primary cause of blindness in the USA. While emergence of ocular inflammation can be genetic or idiopathic, pathogens such as bacteria, fungi and viruses that infect the eye are primary causes. Microbial infections are treated with antimicrobials followed by anti-inflammatory drugs such as dexamethasone, prednisone, cyclosporine etc. The treatment regimen for these anti-inflammatory drugs is prolonged and can last up to many months or even years. While systemic dosing is often avoided unless necessary, topical dosage regimens can have low patient compliance due to repeated administration requirements per day. One such ailment that affects millions of people around the globe is ocular herpes. Primary infections are usually seen in children or adolescents and associated with vesicular dermatitis, follicular blepharo-conjunctivitis, superficial punctate keratitis (SPKs) or dendritic ulcer with preauricular lymphadenopathy. However, herpesviruses can reside forever in the host latently and cause recurrent ocular infection in patients of all ages. While the most common treatment for treating active ocular herpes infection includes daily dosing of Acyclovir (ACV) or ACV analogs, curbing post viral inflammation, which can cause ocular pain and permanent clouding of the cornea, is performed by topical glucocorticoid treatments such as prednisone or dexamethasone. These topical glucocorticoid therapies are avoided during active viral replication and only prescribed in conjunction with oral antivirals to avoid the risk of involving deeper stromal structures with threats to vision. While topical glucocorticoid treatments are effective in reducing inflammation in most cases, they require repeated dosing (6-10 times daily) and suffer from poor ocular retention. A simple yet effective way to improve ocular retention time for topical therapies is to use sustained drug release platforms. We have recently shown that ACV loaded into highly porous activated carbon termed as drug encapsulated carbon (DECON), can effectively adsorb to the corneal surface and deliver drug in a sustained fashion. We showed that use of DECON to deliver ACV, reduced the dosage frequency from 3 in a single day to a single dose once every alternate day in mice. In this regard, we propose to demonstrate a new use for DECON as a sustained drug release platform to deliver dexamethasone during post viral inflammation period. In this proposal, we would like to (1) understand the efficacy of drug loading, retention and release of dexamethasone from DECON using in vitro, ex vivo and in vivo models and (2) through clinically relevant metrics assess DECON’s ability to reduce infectious and non-infectious suppress inflammation during post viral replication period.

NIH Research Projects · FY 2025 · 2023-09

Abstract Sexually transmitted infections (STI) continue to be a major public health problem for Black girls in the United States. Each year 1 in 4 Black girls, 14-19 years-old acquires an STI, placing them at risk for poor sexual and reproductive health outcomes (SRH) (i.e., pelvic inflammatory disease, infertility, HIV/AIDS). In Chicago, STI rates are highest among 13- to 29-year-old Black girls and they represent 56% of new HIV diagnoses, compared to other racial groups, making adolescence an exceptionally vulnerable period. These racial disparities require new and innovative strategies to reduce Black girls' negative SRH outcomes. Familial protection is seen as critical to mitigating risk, particularly exposure to sexual violence which is linked to girls' HIV/STI risk. Interventions that strengthen family relationships and communication as strategies to protect Black girls have demonstrated success improving Black girls SRH outcomes. Yet, with few exceptions, these programs engage only female caregivers, whereas male caregivers may amplify the protective effects of families on Black girls' SRH. We systematically adapted IMARA (an evidence-based program designed for Black girls and their female caregivers) to create IMARA for Black Male caregivers and Girls Empowerment (IMAGE), adding drivers of structural violence (i.e., stereotype messaging and lack of protection) aligning with the Becoming a Sexual Black Woman framework and the Health Disparities Research Framework. Preliminary data (interviews, focus groups, theatre, and pilot testing) with Black girls, male and female caregivers justify the proposed randomized control trial (RCT). We will simultaneously conduct an effectiveness RCT with individual- level randomization and examine implementation processes at five community-based organizations. Aim 1 is to conduct a 2-arm RCT (IMAGE vs. a health promotion control) with 372 14-18-year-old Black girls and their male caregivers and compare girls' sexual risk behavior (condom use, sexual debut, and sexual partners) and STI incidence at baseline, 6- and 12-months. We hypothesize Girls in IMAGE will report more condom use, later sexual debut, fewer sexual partners, and have lower STI incidence at 6- and 12- months (primary outcome) compared to the control group. Aim 2 is to examine change in the theoretical mechanisms posited by the Becoming a Sexual Black Woman. Aim 3 is to identify processes, barriers, and facilitators associated with primary outcomes to inform future implementation into community-based organizations. The long-term significance and impact of this application is high. By including Black male caregivers in the protection of girls, this study leverages a long-neglected yet important resource in Black girls SRH, thereby amplifying the protective effects of family-based programs and pushing the science of health disparities forward.

NIH Research Projects · FY 2024 · 2023-09

ABSTRACT Pregnancy is a complex state that involves immune crosstalks between maternal cells in the decidua and fetal cells. This communication plays an important role in protecting the fetus from rejection. Extravillous trophoblasts (EVTs) are critical fetal cells that shape the immunological microenvironment at the maternal-fetal (M-F) interface. EVTs express a unique set of major histocompatibility complex (MHC I) molecules on their surfaces: the classical HLA-C and non-classical HLA-E, -F, and -G molecules. Quite exceptionally, EVTs do not express HLA-A and HLA-B. HLA-C, -E, and -G molecules have been associated with mechanisms of immune tolerance at the M-F interface, and whether HLA-F functions similarly is less well understood. HLA-F is unique among MHC I molecules in several ways: 1. HLA-F exists in more than one molecular form, with and without associated peptides and b2m; 2. HLA-F binds peptides that are unconventionally long, ranging from 8mers to more than 20mers; and 3. Inhibitory and activating receptors on natural killer (NK) cells can distinguish between peptide-filled and peptide-deficient HLA-F molecules. It is therefore logical to propose that a role for HLA-F in M-F immune crosstalks involves interactions between HLA-F expressed on EVTs and NK receptors on decidual NK cells. This is reinforced by the knowledge that HLA-F expression on EVTs is most abundant in early pregnancy, and that NK cells constitute the largest population of maternal immune cells in the decidua during the first trimester of pregnancy. Thus, to develop an understanding of how HLA-F functions as an immunoregulatory molecule, it is critical that we have a strong understanding of the peptide binding properties of HLA-F and the mechanisms by which peptides modulate interactions with NK cell receptors. This R21 application is an early-stage investigation that aims to fill unresolved gaps in our knowledge of HLA-F immunobiology. In two aims, we propose to examine the unique landscape of HLA-F peptides in relation to specialized aminopeptidases that normally generate MHC I immunopeptidomes inside cells, as well as address key questions centered on why HLA-F presents peptides of extraordinarily long lengths, what biochemical and structural properties of HLA-F support binding of long peptides, and what is the role of peptides in modulating HLA-F interaction with NK cells. For this, we will use a combination of cell-based, biochemical, and structural approaches. Upon completion of this project, we will have generated new knowledge on HLA-F that will help understand how its expression on EVTs and engagement with receptors on decidual NK cells, can support a critical regulatory role in immune crosstalks during the early stages of pregnancy. Given that pregnancy complications are often associated with a loss of immune tolerance mechanisms, a characterization of HLA-F as proposed here is important for stimulating new ideas in managing and decreasing the risks associated with such complications.

NIH Research Projects · FY 2025 · 2023-09

Many cellular processes use phase separation to sequesters biomolecules into membraneless organelles or condensates. This concept is of particular importance in the cell nucleus. Most nuclear condensates are associated with specific chromatin loci. Thus, chromatin organization and biomolecular condensate formation are closely related. Topology, epigenetic modification, or enzymatic activity are possible links between the chromatin scaffold and protein condensation. We have previously described condensates that concentrate transcription machinery at super-enhancers but understanding of cell-to-cell variability and locus specific features that nucleate condensates is missing. Preliminary data indicates that active chromatin decorates the surface of transcription condensates below the resolution limit of conventional microscopy. Making use of recent developments in multiplexed super-resolution microscopy of both chromatin elements and protein factors, we will characterize the nature of the chromatin enrichment layer and determine if it reflects the condensate surface as an active interface that regulates transcription. We will for the first time directly observe the interplay of local chromatin topology, epigenetic signatures, and condensate formation in a specific model locus in mouse embryonic stem cells. By scrutinizing with genomic resolution how each of the regulatory elements in the locus interacts with the condensate we will determine if chromatin structures such as enhancer hubs nucleate condensates, or if condensates induce specific chromatin topologies by connecting active elements. Finally, we will investigate at the molecular level how condensate constituents interact with the microenvironment and associated chromatin elements, and test the hypothesis that retention of transcription factors boosts binding site occupancy and stabilizes condensates at regulatory chromatin. Targeted perturbation and locus-specific observation will allow us to answer this question and begin to identify genome-wide rules that predict where precisely chromatin-supported condensates form. The overall vision for this research program is to develop an integrated view of chromatin organization and condensate formation. Experimental data will inspire new conceptual frameworks and help us integrate the full complexity of condensates into paradigms of biological function such as transcription regulation. We expect that new biophysical models of self-organization and self-regulation will emerge from our studies of intracellular condensates.

NIH Research Projects · FY 2025 · 2023-09

Health information is siloed and often suffers from the sheer volume and the heterogeneous nature of complex datasets. This is in stark contrast to what a physician does when a patient is evaluated. Data from electronic health records, blood tests, imaging, electrical recordings, genomic studies, and neuropsychological measures are combined in the clinician’s mind to find the best diagnosis and treatment plan. Nowhere is this more challenging than for studies of the human brain. We have developed a one-of-a-kind human brain tissue bank where each brain sample is spatially linked to a comprehensive multimodal dataset using standardized tissue processing techniques using a home-grown software platform called INTUITION. Highly curated and integrated data are assembled from patients to help both in clinical care and provide a unique research platform for the human brain that is linked to fresh human brain tissues samples. Many patients undergo complex, multistep brain surgeries to treat their seizures. Studies using these standardized tissue methods and INTUITION software have already led to important new discoveries including molecular (genomic, proteomic, metabolomic) biomarkers, imaging methods, brain network discovery, and therapeutic targets that would otherwise not have been identified. The major goals for this collaborative innovation award are to adapt, modify, and expand INTUITION and a brain tissue collection educational program into a standardized, virtual tissue bank and federated data platform for dissemination through 3 CTSA hubs with teams at 5 collaborating institutions in the city of Chicago. The platform leverages common data elements and will use a team of experts to develop new ones. As part of this ambitious project, we will optimize brain image visualization and EEG tools, work with a patient population that spans the age spectrum (pediatric and adult) and develop user-friendly tools that will be sustainable because of their combined utility for clinical care and research. Given the extraordinary value of highly curated fresh human brain tissues and the fact that brain surgeries are becoming less invasive, it is important that we take full advantage of these priceless tissues as soon as possible. We will develop a hands-on education program followed by web-based training modules for human brain tissue localization and processing in parallel with standardized data processing, storage and upload methods for images and EEG studies at each of the 5 sites. This is especially important for rare brain disorders that require collaboration from many centers to create a standardized, virtual pool of tissue samples and associated data.

NIH Research Projects · FY 2025 · 2023-09

Abstract. The neural substrates underlying alcohol use disorder (AUD), remain poorly understood in part due to lack of translational models that recapitulate phenotypes from the human condition. Biosynthesis of the GABAergic neurosteroid, allopregnanolone (Allo) in corticolimbic neurons, regulates stress sensitivity and induces a potent anxiolytic action. In a rodent model of chronic intermittent ethanol (CIE) exposure, decreased expression of Allo biosynthetic enzymes, 5α-reductase type I (5α-RI) and 3α-hydroxy-steroid dehydrogenase (3α-HSD) is associated with Allo level downregulation in the hippocampus (HIP) and cerebellum. Consistently, in AUD postmortem brain, cerebellum Allo levels and neurosteroidogenic proteins and enzymes, such as the translocator protein (TSPO), 5α-RI and 3α-HSD expression decreased in association with aberrant epigenetic marks. Alcohol-induced epigenetic modifications (e.g., DNA hyper/hypomethylation) on transcriptomics and their impact on neurosteroidogenic gene expression, neurosteroid levels, and anxiety are poorly understood. Allo biosynthesis can be upregulated in brain areas that modulate anxiety and alcohol reward by stimulating the epigenetically modifiable nuclear receptor, peroxisome proliferator-regulated receptor (PPAR)-α by the endogenous modulator, palmitoylethanolamide (PEA). Intriguingly, chronic alcohol exposure decreases PPAR- α expression, while stimulation of PPAR-α by PEA decreases both anxiety and alcohol intake. The molecular mechanisms underlying these effects remain unclear. Our preliminary and published results suggest that alcohol-induced aberrant regulation of PPAR-α may affect anxiety via decreasing Allo content. Hypothesis: Chronic alcohol exposure alters methylation/demethylation dynamics that downregulate corticolimbic PPAR-α expression and allopregnanolone biosynthesis, and elevated anxiety. In male and female rats, we will: (AIM 1) Examine the effect of 14-day CIE exposure (EtOH), ethanol acute (24h, W24h) and protracted (7 days, W7d) withdrawal on the epigenetic regulation of PPAR-α expression and downstream effects on neurosteroidogenic enzyme expression; (AIM 2) Investigate the effects of CIE exposure and ethanol acute and protracted withdrawal on the brain content of Allo; and (AIM 3) Study the pharmacoepigenetics of PEA and fenofibrate on Allo biosynthesis and anxiety after CIE exposure and acute and protracted ethanol withdrawal. This study may unveil CIE-induced neurobiological alterations and suggest treatment targets for alcohol withdrawal symptoms.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY In 2009 two laboratories independently discovered the rostromedial tegmental nucleus (RMTg) – a small GABAergic nucleus that encodes negative reward prediction error via the inhibitory control it exerts over midbrain dopamine neurons. Together with subsequent studies demonstrating a role for the RMTg in responding to aversive stimuli, this work sparked new research investigating involvement of this region in the neurobiological mechanisms underlying alcohol use disorder. The medial prefrontal cortex (mPFC) plays a key role in regulating alcohol seeking and taking behavior. Interestingly, our recent work revealed the presence of dense input from the mPFC to the RMTg that spans both the prelimbic (PL) and infralimbic (IL) subregions. Studies manipulating PL and IL activity at the subregional level suggest that these two regions exert opposing control over alcohol seeking. However, the application of circuit-specific approaches is beginning to suggest a more complex picture with differing roles across discrete cortico-subcortical projections. Combined with only a very superficial understanding of the heterogeneity / uniformity of RMTg neurons, these conflicting data make it difficult to develop hypotheses regarding circuit motifs in RMTg and the potential synergistic or dichotomous functions of RMTg-projecting PL and IL mPFC neurons. Given the impact that recent data demonstrating the previously unappreciated heterogeneity of the ventral tegmental area has had on functional understanding of this region, a rigorous investigation of the RMTg is warranted. The aims described in the current proposal will combine circuit- specific labeling with state-of-the-art connectomics approaches to characterize the effects of chronic ethanol exposure on the RMTg and its cortical afferents at nanometer resolution. Acquisition of an RMTg connectome will provide crucial insight into cell structure and synaptic patterns as well as a precise neuronal map of dense cortical input to RMTg thought to play a critical role in top-down control over alcohol seeking. The resulting circuit data will provide a foundation for future studies investigating the functional consequences of dependence- induced synaptic reorganization by integrating subcellular anatomical findings with physiology and behavior.

NIH Research Projects · FY 2025 · 2023-09

Many preschool-age children with developmental delays and disabilities face significant challenges accessing early educational and therapeutic supports that foster healthy development and improve later academic, economic, and health outcomes. Studies also indicate when children receive these needed services, their parents benefit with improvements in mental health, stress, and quality of life. The scientific objectives of this proposal are to 1) test effective strategies to increase access to early childhood special education (ECSE) services for PCw/DD within pediatric clinical settings serving lower-resourced communities, and 2) characterize potential mechanisms by which such strategies may lead to improved multilevel health outcomes. Preschool and Me (PreM) is a novel community clinical linkage (CCL) designed for pediatric settings where families often experience fragmented or delayed connections to developmental services. Combining key components of CCLs with a personalized medical-education care plan and remote navigator support, PreM targets different levels of influence impacting therapeutic access identified in our prior research. In pilot testing, PreM was feasible, acceptable to families, and demonstrated preliminary efficacy on completed ECSE evaluations. Thus, this R01 application proposes to utilize a hybrid effectiveness-implementation approach to test PreM in two real-world service delivery models. Participants (n=320) will be randomized to either: 1) 6 months of PreM (intervention group) or 2) a waitlist control arm beginning the intervention after a 6-month delay. We will follow all participants for 12 months with data collection occurring at 4 timepoints (baseline, 3-, 6- and 12-months). Our specific aims are to test effectiveness of PreM on indicators of ECSE access (primary) and child-, parent-, family- and health service outcomes (secondary); examine theoretically derived mediators of intervention effects using a mixed methods approach; and explore social variables that may influence variability in intervention effects. We will also simultaneously conduct a mixed methods implementation evaluation focusing on implementation outcomes to serve as indicators for implementation success; measures of implementation quality; and intermediate outcomes to understand and address successes and failures in relation to clinical outcomes. The results of this project have the potential to: 1) advance scientific knowledge about how gaps and delays in educational and therapeutic services impact health outcomes among PCw/DD and their families; 2) identify mechanisms to reduce delays in developmental support; and 3) support effective implementation of educational-clinical linkage models within pediatric systems of care which can be utilized to improve health outcomes for families and their children with a range of health conditions.

NIH Research Projects · FY 2025 · 2023-09

Abstract Inflammatory bowel diseases (IBD) occur when there is an unfortunate combination of microbial dysbiosis and genetic susceptibility. Downregulation of Vitamin D receptor (VDR), a host factor, promotes the severity, extent, and duration of mucosal inflammation and dysbiosis. However, most studies examining gut microbiota have primarily relied on fecal or colonic luminal samples. In contrast, few studies have considered the roles of the small intestinal microbiome. Classically, Paneth cells located in the small intestine are a significant source of antimicrobial peptides (AMPs) and proteins important in host defense. Although Paneth cells are located in the small intestine, AMPs are released to the entire intestine, thus shaping the gut microbiome. VDR regulation of gut bacterial pathogenesis has become an emerging area in IBD, however, the aspect of small intestinal microbiome and metabolites has yet to be explored. Given the challenges in treating patients with Crohn’s disease and the limited studies on the small intestinal microbiome, it is critical to understand how Paneth cell VDR is involved in microbial homeostasis, balanced metabolites, and innate immunity. The objective of our current R01 proposal is to study regulatory mechanisms of Paneth cell VDR and to restore microbiome / metabolites in inflamed states. Our preliminary data showed Paneth cell VDR conditional knockout (VDRΔPC) leads to dysbiosis and susceptibility to Salmonella-induced colitis. We established a method to purify Paneth cells and study their alertness to bacterial pathogens. Furthermore, conditional VDR deletion severely changed the metabolite profile. However, the complex mechanisms in the ileitis through the Paneth cell VDR are still unknown. We hypothesize that VDR deficiency in Paneth cells alters the microbiome/metabolites and makes the host susceptible to chronic intestinal inflammation (e.g., ileitis). We have designed two Aims to rigorously examine the hypotheses at the cellular and microbiome levels: Aim 1. Define the mechanism by which VDR maintains healthy Paneth cells and impacts ileitis inflammation. Aim 2. Investigate the role of Paneth cell VDR in altering the microbiome / metabolites and define the mechanism of VDR in restoring microbial homeostasis, when administering microbiome via fecal transplantation or recolonization in originally germ-free mice. Specifically, we will use novel animal models, organoids from human IBD biopsies, and statistical and bioinformatic tools to understand the host factors and aspects of microbiome / metabolites in chronic intestinal inflammation. Our research team includes experts in the following areas: epithelial biology, animal models, clinical gastroenterology, and microbiome. Our studies are innovative because we provide a unifying hypothesis that can potentially account for defective a host factor in VDR signaling pathway, abnormal Paneth cells, and dysbiosis in IBD. Cutting edge technologies and models will be employed to gain greater mechanistic insights into the interrelationships among VDR, microbiome, and metabolites in inflammation. This study will fill the knowledge gap and provide novel treatment by targeting microbiome / metabolites in IBD and other disorders.

NIH Research Projects · FY 2025 · 2023-09

Project Summary Niemann-Pick Type C (NPC) is an autosomal recessive, lysosomal storage disorder with progressive neurodegeneration and no FDA-approved therapy. Over the past decade, our research team has dedicated efforts to establishing a robust Natural History Study along with efforts to identify biomarkers to enhance our understanding of the disease. In this project, we will leverage our complementary expertise to define a catalog of biomarkers to facilitate future clinical trials. The biomarkers that we will characterize will ultimately provide information on disease progression, prognosis, are associated with commonly affected pathways in NPC and will predict therapeutic response. Towards this goal, we will 1) prioritize and validate biomarkers that reflect neuropathological alterations, 2) identify and characterize proximal biomarkers of NPC1 then 3) investigate multiple biomarkers, representing different aspects of NPC1 neuropathology, to provide prognostic information that can be used in parental counseling and to stratify this heterogeneous disorder for therapeutic trials. Our research team’s extensive discovery efforts have revealed twelve candidate markers. Within the scope of this project is our short-term goal to evaluate these markers in a larger cohort of biospecimens using custom developed and easily transferrable methods. Our long-term goal is to utilize these biomarkers as indicators of response in future clinical trials.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT End stage kidney disease (ESKD) is among the top contributors to mortality in the US population. Nearly 40% of ESKD is caused by diabetes, with a natural history that includes three transitional stages: 1.) Development of diabetes; 2.) Initiation of diabetic kidney disease (DKD); and 3.) Progression of DKD to end-stage disease. Despite the public health burden posed by ESKD, interventions to preserve kidney function in patients with diabetes are limited. Black and Hispanic groups face higher burdens of diabetes and more rapid progression to ESKD than White, non-Hispanic groups. Social determinants of health (SDOH) and other environmental exposures (e.g., metals) are important contributors to these disparities. However, the mechanisms linking environmental exposures to DKD are unclear. Research to integrate environmental data into the multi-omics framework is needed, particularly in Black and Hispanic communities, who suffer from excessive ESKD and are burdened by life-long, adverse environmental exposures. Our goal is to establish a diabetes and kidney disease study site (DSS) comprised of 300 racially and ethnically diverse adults, including 200 with diabetes (half of whom also have kidney disease) and 100 healthy controls. Our DSS will be part of a collaborative initiative to advance the application of multi-omics technologies to study health and disease in ancestrally diverse populations. We will actively engage with this consortium to develop generalizable study protocols, ranging from participant recruitment to integrative analytic pipelines that can be shared and deployed in the cloud. To successfully recruit 300 study participants (Aim 1), we will leverage our established recruitment infrastructure, utilizing an innovative selection strategy that will enrich our sample for those most likely to transition across DKD stages. Thirty DKD cases will be dually recruited with the UIC Kidney Precision Medicine Project (KPMP), enabling linkage to rich KPMP kidney tissue histopathology and multi-omics data in a subsample of DKD cases. Our sample will reflect the diversity of our health system, which includes a patient population that is predominantly non-White (~80%). We will further leverage our extensive clinical research experience to collect biospecimens and obtain detailed information on environmental exposures, outcomes, and other covariables annually for three years (Aim 2). Collected blood specimens will be used to carry-out genomic, epigenomic, transcriptomic, proteomic, and metabolomic profiling among all study participants (Aim 3). Utilizing pipelines and integrative analytic protocols developed in collaboration with the consortium, we will identify molecular profiles linked to environmental exposures and kidney histopathology and examine their associations with each stage of the DKD course (Aim 4). We expect our DSS to have important research impacts, contributing critical information towards the general advancement of integrative systems biology research methods and elucidating novel biological mechanisms and biomarkers for DKD.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT Antigen receptor genes are assembled from several gene segments via V(D)J rearrangement during early lymphoid cell development to generate a diverse repertoire of antibodies. During early B cell development in the bone marrow (BM), V(D)J and VJ joining occurs on the IgH and L chain genes, respectively and is mediated by the RAG recombinase. VH genes are dispersed through 2.5 Mb of the Igh locus. Locus compaction serves to facilitate spatial proximity between the rearranged DHJH join and distal VH genes. Furthermore, V genes rearrange with very different intrinsic frequencies. However, little is known about the precise looping structure of the Igh locus that leads to locus contraction. We undertook an analysis of the entire Igh locus using chromosome conformation capture (3C) based methodology to systematically characterize three-dimensional (3D) chromatin organization on several genomic scales. We found that the Igh locus is compartmentalized into a topologically associated domain (TAD) that is partitioned into three sub-domains. A set of pro-B cell- specific very-long range looping interactions bridge the sub-domains and are Pax5-dependent. These looping interactions are anchored at Sites I, II, II.5 and III and which are critical facilitators of Igh locus contraction. We have now defined the DNA motifs in these loop anchors and discovered a series of pro-B cell specific novel enhancers (NEs) that participate in a NE-NE-VH gene promoter interactome. We have systematically characterized locus compaction using specific KO mice in combination with chromatin-loop mapping methods and newly constructed NE1 KO mice and cell lines. To begin to understand NE interactome function we asked whether those NEs engaged in E-E and E-Pr looping are in an active state as defined by the H3K27Ac histone marks in single cells. We discovered that the NEs are marked by remarkably large H3K27Ac foci. Here we will 1) systematically characterize NEs individually and in the NE interactome as it relates to VH gene usage during repertoire formation, and 2) examine the relationship between the NE interactome and H3K27Ac foci. The presence of H3K27Ac foci on NEs may indicate the participation of the Igh locus in transcriptional condensates which may define the molecular environment for V(D)J recombination.