Pennsylvania State Univ Hershey Med Ctr

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract Genomic stability is critical for cellular function, however, postmitotic cells such as highly metabolically active neurons face the biggest challenge as it must maintain its genome over organismal lifetime. DNA damage increases with age and is accelerated in Alzheimer’s disease (AD) and multiple neurodegenerative disorders. Hence specialized DNA polymerases have evolved to repair different DNA lesions that destabilize DNA helical structure and obstruct replication and transcription. The Y-family polymerases are unique in bypassing DNA damage and synthesizing DNA past specific lesions, thus recovering replication in dividing cells such as DNA Pol kappa (Polk) that can bypass DNA lesions in both S and G0 phase. Surprisingly, we observed high levels of Polk expression in non-dividing differentiated neuronal nuclei in the mice brain that mislocalizes to lysosomes with age and in mice with AD associated transgenes. Almost nothing is known about Polk in neurons and its role in the protracted aging process. Our long-term goal is to understand the role of Polk in combating the DNA damages caused by cumulative exogenous and endogenous stressors, maintenance of the central nervous system (CNS) genome and its relationship with AD. Here we will investigate Polk's role in neuronal nuclei, and how mislocalization of Polk impacts neuronal maintenance during aging and in AD. This will open a novel line of investigation of specialized Y-family DNA polymerases in age associated neuronal disorders like Alzheimer’s. Our central hypothesis is that Polk assists in multiple DNA damage response pathways to prevent genomic instability and combat constant accumulation of DNA damage in post-mitotic neurons, where its expression is critical in the neuronal nuclei. However, with aging and age-associated neuropathy decline in Polk's expression in the nucleus and concomitant accumulation in the lysosomes results in neuronal genomic instability. We will leverage our proteomics data to identify Polk associated proteins that work together as DNA damage response to sustain neuronal genomic stability. Unbiased proteomics in neuronal nuclei will be performed to identify novel Polk interactors. Overexpression of Polk in aging and AD genotype mice will investigate causal role of Polk in neuronal genomic stability. Specific Aim1 will test the hypothesis that Polk is associated with distinct DNA damage response pathways in neurons and identify novel interactors of Polk Specific Aim2 will test the hypothesis that recovering Polk's expression in neuronal nuclei can rescue genomic stability in aging and AD neurons

NIH Research Projects · FY 2026 · 2024-09

Abstract Alcohol use disorder (AUD) is the third leading cause of preventable death in the U.S. and 32.6 million adults have AUD. AUD etiology is highly diverse and current treatment strategies for AUD have high rates of relapse (40 to 60%). Therefore, identifying novel factors facilitating development of AUD is critical, and expansion of the `tool-box' beyond current behavioral and pharmacological targets is required for development of novel effective treatments. A critical gap that hampers discovery of novel AUD treatment is the lack of studies examining the interaction of the brain and body during various stages of the disease. The vagus nerve is the main conductor of brain-body signals and previous clinical and preclinical evidence, as well as our preliminary data, suggest dysfunction of vagus nerve activity is a driver of AUD development and relapse risk. Furthermore, our preliminary data shows that a selective gastric vagotomy (VX) results in increased alcohol intake and preference in rodent models and is associated with alterations in paraventricular (PVN) hypothalamic neurocircuits. Therefore, we propose to use a combination of behavioral, electrophysiological, molecular, histological and viral transgenic techniques to test the novel hypothesis that disruptions of the gastric vagus nerve activity may increase alcohol preference via selective down-regulation of PVN oxytocin function. As a corollary to this, we will investigate whether targeting PVN oxytocin neurons may rescue VX- induced increases in alcohol preference. Specific Aim 1 will test the hypothesis that VX will increase alcohol reward (preference and seeking) and alters PVN gene expression. Specific Aim 2 then will test the hypothesis that activation of PVN oxytocin neurons will rescue increased alcohol preference following VX. Identifying specific brain targets and mechanisms underlying the beneficial effects of healthy vagal activity to reduce alcohol preference, thus reducing motivation for alcohol consumption, could further aid development of both novel pharmacological and non-pharmacologic AUD interventions.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Spatially and temporally precise Ca2+ signals control a vast spectrum of cellular functions including contraction, secretion, transcription, and growth. These signals are mediated by the coordinated function of Ca2+ sensors and Ca2+ channels. STIM proteins in the endoplasmic reticulum (ER) are luminal Ca2+ sensors in the endoplasmic reticulum (ER) that physically couple to highly Ca2+-selective plasma membrane Orai channels, through an extraordinarily dynamic inter-membrane ER-PM junctional coupling process, to generate "store-operated" Ca2+ signals. Dysregulation of STIM/Orai expression is associated with a range of disorders including renal fibrosis, Idiopathic pulmonary fibrosis, immune deficiency, muscle weakness, skin dysplasia, and cancer. The proposed studies address the crucial unsolved questions of how STIM proteins undergo activation, what is the molecular basis of coupling between the distinct STIM and Orai isoforms to generate Ca2+ signals, and what is the pathophysiological role that the STIM/Orai pathway in cellular remodeling in disease, in particular, in the mediation of fibrosis, a major feature in multiple chronic diseases leading to global morbidity. Understanding this fundamental signaling mechanism provides critical new information to generate pharmacological tools to modify Ca2+ signals and alleviate diseases of cellular growth and remodeling. Genetic alterations giving rise to gain or loss of function of STIM and Orai proteins lead to severe immunological, muscular, skin, and neural deficits in humans, and provide critical molecular insights into the structure/function of STIM/Orai proteins. To deepen our understanding of this critical signaling pathway, the project goals focus on: (1) Investigating the mechanisms underlying STIM protein activation by Ca2+ store-sensing or temperature change, and the precise molecular coupling of STIM proteins to Orai channels, focusing on the highly distinct STIM and Orai isoforms, and utilizing precise molecular/genetic probes, imaging technology, and gene-deleted cell lines and animal models: (2) Studying how the small molecule 2-aminoethyldiphenyl borate (2-APB) specifically modulates STIM/Orai isoforms, using genetically-encoded and optogenetically/chemogenetically applied Ca2+ probes tagged onto STIM and Orai proteins to monitor cytosolic Ca2+ signals mediated by STIM/Orai, and exploring how 2-APB provides crucial information on STIM-mediated clustering of Orai channels and controls local junctional Ca2+ signals, advancing our mechanistic understanding and the development of small molecules targeting the STIM/Orai pathway; (3) Examining how STIM/Orai-mediated calcium signals regulate fibrosis in both cellular and animal model systems, focusing on how the STIM1/Orai2 isoform pathway can be modified to regulate the process of fibrosis, determining transcriptomal changes related to STIM-specific Ca2+ signal generation, and utilizing a novel STIM-specific inhibitory peptide, Orai3-M4x, to regulate cell remodeling. Overall, the program of study seeks to understand the fundamental STIM/Orai signaling pathway that has enormous potential in alleviating a spectrum of pathological states mediated by cellular growth and tissue remodeling.

NIH Research Projects · FY 2024 · 2024-09

ABSTRACT: Left ventricular non-compaction (LVNC), a rare but increasingly prevalent cause of cardiomyopathy stemming from developmental arrest of myocardial compaction, was identified as a primarily genetic, independent cardiomyopathy by the AHA in 2006 following debate due to its frequent association with variable degrees of myocardial dysfunction and other congenital heart defects (CHD). Symptom heterogeneity has likely left many cases undiagnosed, giving a lower estimation of its real impact as a cardiomyopathy. Congenital LVNC can progress in severity throughout life contributing to decompensated heart failure and sudden cardiac death. Little is known about the molecular mechanism(s) that lead to LVNC, but mutations in MYH7 (encoding human beta- cardiac myosin) have been implicated as a putative genetic cause. Human beta-cardiac myosin (M2β) mutations are known molecular triggers for heritable cardiomyopathies, typically impacting the motor protein’s intrinsic motor properties and/or auto-inhibited state. Studies of cardiomyopathy mutations in M2β have demonstrated that hypertrophic cardiomyopathy (HCM) is triggered by mutations that cause hyper-contraction and dilated cardiomyopathy (DCM) is triggered by mutations that cause hypo-contraction. However, very few studies have examined LVNC mutations and thus it is unclear how it falls on this genotype-phenotype spectrum. Examining LVNC cardiac myosin mutations would enlighten us about the unique etiology and reveal important information about myosin structure and function. We hypothesize that depressed monomeric ensemble force and/or an increase in autoinhibited state stabilization could trigger severe hypo-contraction and tissue level tension/force imbalances, leading to excessive trabeculation in LVNC. We conclude that additional biophysical LVNC M2β mutant studies both in purified protein and cellular systems are required to uncover common allosteric pathways altered in LVNC. Our goal is to investigate the impact of clinically identified MYH7 LVNC mutations M362R and L655M on protein structure-function, mechanochemistry, and ultimately pathogenic myocardial remodeling. M362R is in the loop 4 region of the motor domain known to play an important role in head-head interactions, tropomyosin positioning, and actin binding. L655M may disrupt a critical allosteric pathway leading to actin-activation of the power stroke and thus impair the transition into the force generating states. This proposal will characterize two LVNC mutations selected based on the isolation of LVNC from other forms of heritable cardiomyopathy and on their respective locations within myosin known to be important for performance. We will utilize transient kinetic, novel applications of FRET biosensors and analytical ultracentrifugation, and hiPSC-CM derived myofibril-based techniques to elucidate how these mutations impair myosin function leading to LVNC pathogenic remodeling of the heart.

NIH Research Projects · FY 2024 · 2024-09

Project Summary/Abstract: The overall goal of this project is to determine the association of mushroom intake with cognitive function in a middle-aged and elderly cohort conducted in Aichi Prefecture, Japan. Healthy diets are associated with reducing the risk of cognitive decline and there is a need to identify specific foods that delay or reduce cognitive impairment in the elderly. Mushrooms are foods that contain several beneficial ingredients that have shown to reduce cognitive decline in animal studies, possibly through anti-oxidant and anti-inflammatory pathways. Mushrooms are infrequently consumed in the United States and their effects in humans have been understudied. We showed in the National Health and Nutrition Examination survey that in persons over age 60 who participated in a battery of cognitive tests, mushroom consumers evidenced higher scores and better cognitive function. Mushrooms are fungi, and contain unique compounds not found in plants and animals. Mushrooms contain high concentrations of ergothioneine, a thiohistidine betaine amino acid that is a powerful antioxidant. In particular, the ergothioneine transporter OCTN1 gene is a specific transporter for the uptake of ergothioneine to cells and tissues experiencing oxidative stress. Mushrooms also contain high concentrations of glutathione (GSH), the most abundant intracellular antioxidants in humans. The traditional Japanese diet, which includes mushroom consumption, is associated with a lower risk of cognitive decline in the Ohsaki Cohort 2006 Study. We have the opportunity to study the individual effects of mushrooms and their key constituents in the Japanese National Institute for Longevity Sciences-Longitudinal Study of Aging. This is a multi-wave cohort study conducted in men and women ages 40-79 who have been followed for age-related health effects including cognitive measures. The study uses detailed questionnaire, medical records, and nutritional assessments. There are several reasons why the NILS-LSA is the ideal population for our hypothesis. Mushrooms are consumed by a higher percent of the population in Japan and in greater daily amounts than in the U.S. In addition, the ergothioneine content of mushroom species consumed in Japan is higher than the white button mushrooms typically consumed in the U.S. Our specific aims are: 1. Determine the relationship between mushroom intake and a battery of neurocognitive outcomes including cognitive function, dementia, and depression. Based on 24-hr food recall data, mushroom intake at baseline and at success waves will be examined prospectively using generalized equation models in relation to a battery of cognitive measures including medically determined clinical dementia, the Wechsler Adult Intelligent Scale (WAIS-R digit span test), the Mini-Mental State Examination, and clinical depression. 2. Determine the relationship between ergothioneine and GSH levels and risk of neurocognitive measures using baseline blood samples for a subset of 309 participants. Causal mediation analysis will be conducted to determine the direct effects of mushroom and indirect effects attributed to ergothioneine and GSH.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT The antiparkinson drug levodopa (L-DOPA) is the current standard of treatment for Parkinson’s Disease (PD) and can be used as either a monotherapy or in combination with monoamine oxidase-B (MAO-B) inhibitors, such as selegiline to ameliorate dopamine depletion. Despite L-DOPA’s recognition as the gold standard for PD management, our group has recently made a fundamental observation of iron accumulation in the substantia nigra (SN) of PD patients following L-DOPA but not selegiline administration. These findings indicate that the type of antiparkinson drug may be correlated with nigral iron increase. Evidence suggests that nigral iron accumulation is increased in PD patients, decreasing dopamine thus exacerbating the progression of neurodegeneration. Though, it remains unclear whether increased iron in the SN is from PD itself or from L- DOPA treatment. My objective for this proposal is to demonstrate that L-DOPA exposure alters the iron status through direct effects on dysregulation at the BBB (Aim 1) and/or through altering the molecular and cellular profile in the SN (Aim 2). Aim 1 of this project will elucidate how L-DOPA exposure will cause an increase in iron transport from the blood-brain barrier (BBB) to the brain in an in vitro BBB model. I will measure iron transport in endothelial cells after drug administration. Aim 2 will examine how L-DOPA delivered orally will impact the molecular, histological landscape, and iron metabolism in the ventral midbrain. Furthermore, Aim 2 will take a unique perspective to study how iron deficiency alters brain iron and exacerbate the severity of iron accumulation in a rat model by examining brain iron levels and immunohistological staining expressions. PD patients have a high prevalence of an anemic diagnosis, however, there is minimal research regarding the effects of L-DOPA and anemia on brain iron homeostasis. This research project will deepen our understanding of how antiparkinson treatments such as L-DOPA and selegiline impact iron homeostasis with the hopes of identifying iron-based mechanisms from the treatment. Importantly, advances in iron supplements will provide novel therapeutic targets to improve our standard of care for PD patients The proposed training plan is sponsored by Dr. James Connor. The overall goal of the training plan is to provide the fellowship applicant with a solid foundation for a successful career as a researcher and enhance techniques for translational studies. The goals included in the training plan are 1.) Gain experience and competence in various techniques integrating iron biology and neuroscience, 2.) collaborate with other research scientists to learn and complement my knowledge, 3.) develop hypothesis-driven research, 4.) conduct translational research, and 5.) effectively present data in a manuscript.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT COPD is the fourth leading cause of death for women in the United States, which has a strong genetic basis. Studies have demonstrated the heritability of COPD to be ~40%. Despite the enormous burden of COPD, there are still no pharmacologic therapies that slow progression of disease or reduce mortality, indicating the need for a better understanding of the genetic basis of the disease. Notably, COPD shows sex differences in disease prevalence, pathology, and symptoms. Although historically thought to affect primarily men, the risk of COPD in women has increased over time due to growing prevalence of smoking and exposure to harmful pollutants. Thus, over the past decade, age-adjusted prevalence of COPD has been consistently higher in women than in men. Moreover, women report greater breathlessness at similar degrees of airflow obstruction and emphysema severity, and experience >25% more exacerbations than men. Despite the sex differences, most omics studies of COPD combine both sexes together and large-scale female specific COPD studies are lacking. In this application, we propose novel approaches to conduct female-specific GWAS, borrowing strength from summary statistics from large sex-combined GWAS of COPD. We will also generate single cell RNASeq data from female lung and integrate cell type specific eQTLs with GWAS to identify risk genes. Besides, we will assemble a large multi-ancestry plasma proteomics dataset, generate female specific pQTLs, and use them to identify COPD biomarkers. Results from this study will bring novel insights into the mechanisms of COPD in females and identify protein biomarkers to aid in the diagnosis.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Merkel Cell Polyomavirus (MCPyV) is a double-stranded DNA human tumor virus that is the main causative agent of Merkel Cell Carcinoma (MCC), a rare, but extremely aggressive skin cancer. MCC predominantly affects elderly or immunosuppressed patients. Currently, there is a lack of durable and effective treatment options to treat this cancer, thereby necessitating the need to identify new therapeutic options as rates of MCC are steadily increasing. Before the onset of MCC, MCPyV establishes a latent infection and therefore, must have its own mechanisms to promote long term survival within its host to cause MCC later in life. It is known that other human tumor viruses, such as Human Papilloma Virus (HPV), induce cell cycle arrest in infected cells to promote viral persistence. My previous results showed that MCPyV induces host cell cycle arrest and cellular senescence by expressing the large tumor antigen (LT) as a mechanism to promote viral genome maintenance in human fibroblast cells. When investigating how MCPyV LT might induce cellular senescence, we observed that this viral oncoprotein induces the nucleolar stress response (NSR), a host stress response that functions to cause cell cycle arrest and/or cellular senescence, usually in a p53-dependent manner. The NSR is canonically activated upon factors that can inhibit ribosomal biogenesis and often results in disruption of the nucleolar structure and activation of p53. Though this stress response has been characterized in response to nutrient starvation, chemotherapeutic drugs, or other stressors, it has not been sufficiently defined in the context of viral infections. The goal of this proposal is to determine how the NSR is regulated by MCPyV infection and how this stress response might impact MCPyV replication. Specifically, my preliminary data indicated that MCPyV LT downregulates processes that regulate ribosomal RNA (rRNA) expression and processing, or dysregulates ribosomal assembly and/or protein synthesis seen by a lack of phospho-ribosomal protein S6 (p-RPS6) expression, implicating a potential role of LT in modulating ribosomal biogenesis on the RNA and protein level. Accordingly, the central hypothesis of this proposal is that MCPyV LT can inhibit ribosomal biogenesis to induce the NSR, which is required for MCPyV viral persistence. To address this hypothesis, Specific Aim 1 will characterize the NSR phenotypes and elucidate the mechanism and processes of ribosomal biogenesis that are directly disrupted by MCPyV LT. Specific Aim 2 will investigate the mechanism by which MCPyV LT promotes p53 activation and how the p53-dependent NSR regulates MCPyV genome replication and persistence. Taken altogether, the findings from this proposal will significantly expand our understanding on the role of the NSR as a critical viral sensor and identify host factors that are essential for MCPyV pathogenesis.

NIH Research Projects · FY 2025 · 2024-08

Project Summary With a fast pace of population aging, the prevalence of cognitive impairment and dementia including Alzheimer’s disease is expected to rise exponentially, placing an enormous social and economic burden worldwide. Epidemiologic and pathological studies have indicated an association between vascular diseases and cognitive dysfunction, implicating vascular contributions to cognitive impairment and dementia (VCID). However, the driving pathologic mechanisms of VCID remain unclear, preventing the development of effective prevention and treatments. In response to the NIH Notice of Special Interest (NOT-HL-23-002): Promoting research to understand VCID, this proposal aims to identify the mechanisms of cognitive impairment caused by atherosclerosis and red blood cell (RBC) released ATP using both animal models and human samples. Atherosclerosis is the leading cause of vascular disease worldwide, yet the role of atherosclerosis in cognitive impairment and the underlying mechanisms remain largely unknown. Our study using high fat diet (HFD)-fed ApoE-/- mice showed that atherosclerotic plaques extensively blocked carotid and intracranial arteries, resulting in brain hypoperfusion, cerebrovascular remodeling and neuroinflammation. Importantly, cognitive behavior tests detected significant memory deficits in ApoE-/- mice as early as 12 weeks with HFD, but not in age matched mice with normal diet, demonstrating a causal relationship between atherosclerosis and cognitive impairment. Our study also showed that disturbed blood flow-triggered ATP release from RBCs via pannexin 1 channel (Panx1) contributes significantly to the initiation and progression of atherosclerosis as well as cognitive impairment. Specifically, Panx1 deletion from RBCs led to reductions of plaque formation and alleviations of the cognitive deficit in HFD-fed ApoE-/- mice. Additionally, we showed that hypercholesterolemia enhances vascular endothelial cell (EC) [Ca2+]i responses to ATP, a key signaling for EC barrier dysfunction. These findings led to our central hypothesis that atherosclerosis-induced brain hypoperfusion and vascular inflammation via hypercholesterolemia along with RBC-released ATP are the key risk factors of atherosclerosis-driven cognitive impairment. Three specific hypotheses will be tested experimentally in vivo, in vitro, and computationally in silico under three specific aims. Aim 1: The atherosclerotic blockage of carotid and intracranial arteries causes brain hypoperfusion and cerebrovascular remodeling, resulting in cognitive impairment. Aim 2: RBC-released ATP contributes significantly to atherosclerosis-induced cerebrovascular pathology as well as the onset and progression of cognitive impairment. Aim 3: Hypercholesterolemia and hemodynamic alteration-induced RBC released ATP are the key risk factors synergistically contribute to atherosclerosis induced cerebrovascular inflammation and EC barrier dysfunction in mice and humans. The new knowledge gained from this study will advance our understanding of the risk factors and mechanistic insight into atherosclerosis-induced VCID and hence promote early treatment and lifestyle interventions to prevent or delay the onset of cognitive decline.

NIH Research Projects · FY 2026 · 2024-08

Project Summary/Abstract: This is an application for a K01 award for Djibril M. Ba, PhD, MPH, an assistant professor at the Penn State College of Medicine in the Department of Public Health Sciences. Dr. Ba is uniquely qualified to conduct this research project based on a strong foundation in nutritional epidemiology for examining diets in relation to disease occurrence. The proposed K01 application will provide Dr. Ba protected time, which will be devoted towards training and career development activities targeting the following objectives to strengthen Dr. Ba's trajectory toward becoming a successful independent investigator: (1) Acquire knowledge and expand didactic learning; (2) Gain advanced quantitative skills required for developing different dietary indices and analyzing longitudinal dietary data from epidemiologic studies; (3) Enhance Dr. Ba translational grantsmanship and leadership skills to effectively compete for future NIH funding; and (4) Obtain the knowledge base and professional skills to perform Data Coordinating Center activities. Dr. Ba has assembled a mentoring team comprised of a primary mentor, Vernon M. Chinchilli, PhD a leading expert in the field of biostatistics and Data Coordinating Centers, and three co-mentors: Dr. Phil Hart (gastroenterologist), Dr. Qi Sun (nutritional epidemiologist), and Dr. Nazia Raja-Khan (endocrinologist). Healthy plant-based and Mediterranean diets, which are based on high consumption of antioxidant-rich foods such as fruits and vegetables, have been associated with a lower risk of diabetes in the general population. In contrast, pro-inflammatory diets such as those reflected by a high empirical insulinemic index and a high empirical dietary inflammatory index including red and processed meats, may lead to an increased risk of diabetes. These dietary components are known to be positively associated with endothelial dysfunction, inflammation, insulin resistance, and oxidative stress that can cause tissue damage, especially reduced pancreatic β-cell function. Almost one-third of patients with acute pancreatitis (AP) will develop diabetes within 3 years but it is unclear whether the risk is based on dietary patterns. This constitutes a critical knowledge gap regarding the effects of different dietary patterns on the prevention of diabetes following AP and whether such risk can be mitigated. The overall objective of this application is to acquire detailed knowledge and novel research skills needed to accomplish the following research aims: Aim 1: To prospectively determine the role of dietary patterns in the development of diabetes (primary outcome) or pre-diabetes (secondary outcome) following an episode of AP; Aim 2: To determine the associations of dietary patterns with weight changes, waist circumference, and biomarkers of metabolic alterations related to diabetes; Aim 3: To determine whether weight changes and biomarkers of metabolic alterations are mediators for the association between dietary patterns and the risk of new-onset diabetes among AP participants in the DREAM study. Findings from this project will provide strong evidence about the role of dietary patterns on the occurrence of future diabetes following AP.

NIH Research Projects · FY 2025 · 2024-07

The HIV pandemic has had an enormous impact on human health, with 39.9 million people living with HIV and 1.3 million new infections in 2023. Despite clear evidence at the couple-level that HIV test-and-treat strategies are successful at reducing HIV transmission, population-level trials have shown mixed effectiveness of test-and-treat strategies to reduce HIV incidence. There is evidence of disparities in testing and treatment engagement. Furthermore, these disparities are frequently along the same characteristics as sexual network clustering. Therefore, a potential explanation for a smaller than expected incidence impact is sexual network clustering, partnering of like-with-like, by HIV testing and treatment. Indeed, my preliminary work showed that couples who reported never testing for HIV were nearly 2 times more likely (1.9, 95% CI=1.8-2.0) to be partnered with one another than expected by random chance. But this finding needs replication, as there is little empirical evidence to support these claims. Through network analyses and simulations, contextualized with community interviews, this career development award will assess whether sexual network clustering impacts the effectiveness of HIV treatment as prevention. These results will inform population-level treatment targets. Further, they will allow me to test the utility of network-driven strategies to drive testing and treatment and provide clear guidance to maximize the impact of these interventions in the USA. Research aims will include: 1) Identify the network context of HIV test-and-treat interventions by (A.) Characterizing the sexual network position of people engaged in test-and-treat, and (B.) Estimating the level of sexual network clustering by test-and-treat; 2) Evaluate the impact of network-driven strategies of HIV interventions using network models parametrized with data on engagement in HIV test-and-treat and sexual network context; 3) Translate modeling and network analyses to real-world settings. Analyses of data in contexts with higher HIV prevalence will allow for efficiency in conducting analyses and drawing inference to domestic settings in the USA. Further, in order to test these hypotheses in the available time, this proposal will leverage previously collected comprehensive existing data: the Rakai Community Cohort Study, the TRUST/RV368 Study, Population-based HIV Impact Assessments, and the Likoma Network Study, and collect original qualitative data. This work will develop my expertise in network science and translation through a mentorship team with extensive experience in network data collection and conducting community-informed work, training which I will apply to conduct future research in the USA. Building on these findings, future funding will be sought to collect expanded sexual network data and develop network-level intervention strategies. My training plan will include gaining experience in participatory modeling and qualitative work, a novel skillset among network modelers. This career development project will provide me with expertise to begin an independent career as a US leader in HIV and sexual network research.

NIH Research Projects · FY 2025 · 2024-07

Stimulant use disorder (STUD) and overdose fatalities are devastating public health problems, straining healthcare systems, and societal productivity. Annually, approximately 55% of stimulant prescriptions have been prescribed to adults, with nearly one-third of them reporting stimulant misuse or stimulant-related harm. Despite growing concern that stimulant medications may increase the risk of STUD and overdose in some adults, risk factors for stimulant-related harm have not been well understood, and no screening tool is available for assessing the risk/benefit of stimulant therapy at the point of care. This project hypothesizes that machine learning (ML) techniques, applied to large clinical (Aim 1) and linked census (Aim 2) datasets, can help identify personalized risk factors for STUD and overdose among adults treated with stimulants, and assist with the development of clinical decision support system (CDSS) tool (Aim 3), which, in turn, can help guide clinical decision making when considering stimulant therapy and decrease stimulant-related risk of harm. To evaluate potential demographic and clinical risk factors for STUD and overdose, retrospective longitudinal ML-based analysis of electronic health records (EHR) from the TriNetX research network will be conducted (Aim 1). Because EHRs do not include characteristics of neighborhoods where patients reside, and the health challenges of the population can contribute to STUD and overdose risk, the predictive model from Aim 1 will be further improved by geocoding and linking the local health system’s EHR to the census block group level neighborhood data for each patient to account for the potential impact of geo-contextual factors and improve the accuracy of the STUD and overdose risk modeling (Aim 2). Risk factors identified through Aim 1 and Aim 2 analysis, along with prescribing-clinician input, will enable the development of a CDSS tool to aid clinicians with risk/benefit assessment of stimulant therapy at the point-of-care, based on unique demographic, clinical and neighborhood characteristics of each patient (Aim 3). The CDSS has been increasingly utilized in digitally- driven healthcare to improve treatment safety and outcomes, but their applications toward improving care for adults treated with stimulants or to mitigate addiction and overdose risks have been lagging. The proposed research leveraging big data and ML has enormous potential for understanding and predicting health risks of stimulant therapy in a personalized way, and will lay foundation for future clinical trials evaluating the efficacy of CDSS-based intervention on reducing the risk, and, ultimately, improving health of adults considered for stimulant treatment. This mentored research scientist development award (K01) will also enable the PI to pursue additional advanced training in relevant clinical knowledge, analytical skills, grantsmanship, leadership, and balanced representation to support his long-term goal of becoming an independent scientist contributing to the mission of the National Institute on Drug Abuse to reduce the burden of substance use and addiction, particularly when it involves stimulants.

NIH Research Projects · FY 2026 · 2024-07

ABSTRACT Autoimmune or immune-mediated diseases affect 10% of the world population, and most conditions have no cure. Existing treatment options are often broadly acting and have numerous side effects. Autoimmune diseases often have a preclinical phase where a subset of symptoms occur, but they usually do not yet meet the diagnostic criteria for the disease stage. Many conditions may also have an early stage where the symptoms are milder. Early diagnosis at the preclinical phase or early disease stage is challenging as the symptoms are usually non- specific. Moreover, only a fraction of individuals will progress to disease stages, which poses challenges for clinical interventions. Understanding the genetic basis for preclinical immune-mediated diseases and dissecting risk factors that underlie the progression from preclinical to disease stage is critical. In this application, we propose to develop a series of methods and software tools to better detect genetic associations with preclinical to disease stage progressions, perform integrative genomic analysis to identify genetic signatures underlie the progression, and dissect genetic contribution to sex differences in autoimmune diseases. The methods and tools will be broadly applicable to a myriad of diseases. The proposed development received enthusiastic support from researchers studying psoriasis, Sjogren's disease, lupus, vitiligo, etc. The proposed new tools can profoundly impact the understanding the genetic basis for preclinical diseases.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Diffuse gliomas are the most common primary malignant adult brain tumor. Lower grade gliomas (Grade 2) inevitably become malignant, and prognosis is fatal. Key clinical challenges include: 1) significant intra- and inter-tumor molecular heterogeneity; and 2) limitations in current imaging techniques for distinguishing true tumor recurrence (TTR), pseudo-progression (PP), or radiation necrosis (RN) following surgical resection and adjuvant therapy. Direct tissue sampling is invasive and may not capture the entire molecular landscape of the tumor. Non-invasive imaging (e.g. MRI) fails to detect small recurrences, is limited in resolving differential diagnoses during imaging follow-up, and does not capture tumor molecular evolution. Recently, liquid biopsy of proximal fluids has gained popularity for systemic tumors. In gliomas, both blood and cerebrospinal fluid (CSF) are viable sources of proximal fluids. While acquisition of blood is less invasive, the CSF is physiologically expected to be a superior enriched reservoir for tumor-specific biomarkers. Thus, CSF can be leveraged for more direct target identification, which can be subsequently extended to blood assays. Most molecular analyses of glioblastoma have focused on the genome, epigenome and transcriptome, but proteomic analyses are also required to understand tumor functional phenotypes like response to therapy. We have conducted shotgun proteomics on as little as 30 µL of CSF from patients with glioblastoma and primary central nervous system lymphoma, detecting a substantial concentration of distinct and identifying differentially enriched pathways for each tumor class. We hypothesize that integration of CSF-based proteogenomic analyses can improve our diagnosis of DGs, refine our management during tumor surveillance and offer insight into potential novel drug targets. We propose to conduct a multi-center study focused on CSF, with the largest prospective cohort established to-date. This project will validate our preliminary findings while establishing novel diagnostic proteomic signatures based on CSF. We will also establish the first ever CSF glycoproteome in DGs, which will be of great value for drug target development. Success in this proposal will have a significant impact on diffuse glioma management through improving preoperative diagnostic precision and subsequent surgical planning, enable early intervention for TTR while eliminating potentially unnecessary surgery for RN, and establish a template for drug target discovery in future studies.

NIH Research Projects · FY 2026 · 2024-07

Project Summary The brain undergoes anatomical and physiological changes with the onset of puberty, of which synaptic pruning and increased myelination in a posterior-to-anterior gradient are hallmarks. These neural changes are reflected in the sleep electroencephalogram (EEG), with sleep intensity and depth sharply declining and sleep integrity plateauing during adolescence. Supported by R01 MH118308, we leveraged sleep EEG data from a population- based cohort of children followed-up as adolescents with which we studied the age-related trajectories of slow wave activity (SWA), sleep spindle density (SSD) and odds ratio product (ORP), three EEG biomarkers of sleep intensity, integrity and depth, respectively. We showed that each followed distinct maturational trajectories, differentially depending upon sex and pubertal development. In addition, we showed that, compared to typically developing youth, those with unmedicated attention/learning disorders had a maturational disruption in SWA during childhood and greater ORP levels in adolescence, while those with unmedicated internalizing disorders showed a lower loss of frontal SWA in the transition to adolescence but greater ORP when medicated with antidepressants. The lack of young adult data did not permit unveiling whether those developmental trajectories were predictive of mental health outcomes at ages 20 to 30, a lifespan period critical for increased severity of psychopathology. Federal research priorities include understanding the developmental trajectories of mental health disorders across the lifespan and their underlying brain mechanisms. Hence, in this application, we propose to study the association of SWA, SSD and ORP with young adult psychopathology by leveraging sleep EEG data from the 15-year follow-up of a unique population-based child cohort, studied under our prior R01 MH118308 for their child (median 9y) and adolescent (median 16y) data and who are being recruited under R01 HL136587 for their young adult data (median 25y). We aim to leverage data from these subjects at age 20 years or older and assessed for their sleep EEG and clinically-meaningful psychopathology outcomes. We will study the longitudinal association between the trajectories of SWA, SSD and ORP with internalizing symptoms and suicidality (Aim 1) and externalizing behaviors and working memory (Aim 2), and how these sleep EEG trajectories may play a mechanistic role in the increased coexistence of internalizing and externalizing psychopathology in young adulthood (Aim 3). Given the known sex differences in brain maturation, we will test whether the associations in Aims 1-3 differ between males and females (Aim 4). This study will help better understand the developmental trajectories of psychopathology as they relate to altered brain maturation and sleep disruption at different life stages. Findings from this study will inform novel biomarkers for clinical trials that aim at improving the pathophysiology and long-term prognosis of diverse forms of psychopathology across the life span by targeting the sleeping brain.

NIH Research Projects · FY 2025 · 2024-06

Every living organism is made up of cells that are organized into tissues, whose micron-scale features define normality and disease. These principles inspire the long-term goal of this project: to enable the quantitative characterization of the Geometry of Life and Disease as a foundation for a comprehensive and integrative understanding of how genes, environment and disease determine anatomical and microanatomical phenotypes. Ideally, the effects of genes and environment on anatomical and microanatomical phenotypes can be assessed across the whole organism. We understand the cellular basis of disease as a result of histology's representation of all cell types and two centuries of application to clinical medicine. However, histology is 2-dimensional, qualitative, subject to sectioning artifact, and samples only ~1-2% of specimens. To facilitate anatomically comprehensive phenotyping across whole organisms, we have developed a unique, state-of-the-art 3D modality (histotomography) based on the principles of X-ray tomography and the creation of large-field, high resolution instrumentation for tomographic, synchrotron-based beamlines. The ability to conduct histological assessment in a whole organism in 3 dimensions promises revolutionary advancements on numerous fronts. We propose to establish the groundwork for large-scale access to synchrotron microCT for biology, including a pilot imaging facility at the Lawrence Berkeley National Laboratory (LBNL) and then at Argonne National Laboratory (ANL) for histotomography, to initiate access to visualizations and histotomograms of whole, centimeter-scale model organisms and tissues, and to establish a basic set of web-based visualization and cross-referencing tools for the scientific community. Cloud-based dissemination of raw and reconstructed image data will be piloted at the National Energy Research Scientific Computing Center operated by LBNL or Argonne Leadership Computing Facility (ALCF) at ANL, to enable users to advance their educational, functional, and scientific goals. Notably, the proposed groundwork will comprise a foundation for computational phenotyping that is more comprehensive, objective, precise, and statistically validated. Broad accessibility to microanatomical resources would enable valuable, new means of model validation, reproducibility, and discovery, and provide a new foundation for a more comprehensive understanding of genetic, molecular, and cellular function in the study of normal biology and disease. We expect the proposed work to lead to unexpected research observations, improved clinical outcomes, enhanced robustness and reproducibility of biomedical research, and enhanced sustainability and human health by advancing the quality and comprehensiveness of global monitoring of environmental toxicity and climate change.

NIH Research Projects · FY 2025 · 2024-06

Project Summary/Abstract Information on drug safety is critical to health care and policy decisions, as treatment recommendations hinge on accurate knowledge of both efficacy and harms. However, assessments of drug safety from individual clinical trials are often underpowered due to insufficient sample sizes and have limited generalizability due to restrictive inclusion/exclusion criteria. Meta-analysis of clinical trials offers a unique opportunity to assess adverse event risks in a large sample size for a broad population, but requires careful consideration of how to handle safety outcomes. Critically, methods that are appropriate for assessing treatment benefits are inappropriate for assessing harms. A key distinction between safety and efficacy data stems from the multifaceted nature of adverse events. There are many types of adverse events, each correlated with different potential risk factors. The severity of these adverse events can vary widely, spanning from mild to fatal. Moreover, many adverse drug effects are infrequent and typically suffer from incomplete reporting. In particular, incomplete reporting of adverse events impacts the ability of systematic reviews to synthesize toxicity data, which can promote a false impression of safety or misinform clinical and regulatory decisions. To address these challenges, we propose to develop novel meta-analytic methods for combining safety data. Specifically, our objectives are to develop meta-analysis approaches that can handle multivariate outcome data, account for the severity of adverse events, and identify potential interactions among risk factors through the following specific aims: Specific Aim 1: To develop meta-analysis methods for multivariate outcomes. Specific Aim 2: To develop methods for meta-analysis with ordinal event grading. Specific Aim 3: To develop a method to identify high-risk subgroups. All methods developed will allow for incomplete reporting and be implemented as easy-to-use software. We will apply the proposed methods to understand the drug toxicity profiles and elucidate risk factors of adverse events resulting from cancer immunotherapy and BTK inhibitors. These findings can be used for risk stratification of patients and to inform potential risk-reduction or monitoring strategies. More broadly, the products of this proposal will promote scientific rigor in summary data integration, allowing appropriate inference on the safety of medical interventions and ultimately enhancing patient care. Our proposed research will establish a new modeling framework that will overcome the limitations of current meta- analysis approaches to synthesize data and information on drug safety. This work directly addresses priorities in the strategic plan of the National Library of Medicine to accelerate discovery and advance health through data-driven research. The new approaches, together with publicly available software, will provide a useful tool for the wider scientific community to conduct more rigorous meta-analysis of safety data.

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY/ABSTRACT Medical device-associated microbial infection arises from pathogenic bacterial adhesion and subsequent biofilm formation on devices. It is known that bacterial intracellular nucleotide second messenger signaling plays an important role in biofilm development. Interference with these nucleotides signaling could provide a novel approach to address the problem of pathogenic biofilm formation on biomaterial surfaces. This application proposes to synthesize small molecule derivates of 4-arylazo-3,5-diamino-1H-pyrazole (named as SP02, SP03, and SP04) and tether them to polyurethane (PU) and polydimethylsiloxane (PDMS) biomaterial surfaces so that the small molecules can interfere with nucleotide signaling and interrupt biofilm formation in a way that will both reduce the formation of biofilms and increase the antibiotic efficacy for treating biofilms that do develop. Through testing 4 clinically relevant pathogenic biofilm bacteria on modified surfaces in vitro and in vivo, a new approach for creation of improved antimicrobial biomaterials for implantable medical devices such as catheters will be developed. The central hypothesis is that “biomaterial surfaces tethered with small molecule derivates of 4-arylazo-3,5-diamino-1H-pyrazole can inhibit and interrupt biofilm formation and growth by interfering with intracellular nucleotide signaling. This leads to disruption of biofilm formation and increases efficacy of antibiotics, therefore making microbial infection treatable using standard antibiotic therapy.” To test this hypothesis, four specific aims are proposed. Aims 1 and 2 will focus on in vitro assessment of the effectiveness of three small molecules and these small molecules bonded PU and PDMS biomaterial surfaces for inhibiting biofilm formation and increasing antibiotic efficacies, as well as addressing biocompatibility. The nucleotide levels and RNAseq will be quantified to determine the effects of small molecules on bacterial intracellular nucleotide signaling. Through these experiments, the molecule tethering approach that leads to the greatest inhibition of biofilm formation will be identified for in vivo studies. Aim 3 will test the antibacterial properties and tissue response to small molecule modified biomaterials using a 7-day subcutaneous infection rat model to validate the findings of small molecule functionalized polymers identified from Aims 1 and 2. To accelerate the application of new approach in medical devices, commercial pediatric central venous catheters will be modified with small molecules. Biofilm formation and antibiotic efficacies will be tested using a total implantable venous access port (TIVAP) in vitro and in vivo in Aim 4. The success of experiments described will allow progression to in vivo studies of other medical devices and large animal studies for preclinical trials, as well as providing important basic science information on nucleotide messenger signaling in biofilm formation and control.

NIH Research Projects · FY 2025 · 2024-06

Project Summary Liver fibrosis in chronic liver diseases (CLDs) such as viral hepatitis, metabolic dysfunction-associated steatotic liver disease (MASLD), and metabolic dysfunction-associated steatohepatitis (MASH), is a significant health problem. The hepatocyte injury during CLDs is followed by replacement of liver parenchyma by fibrotic tissue with excessive deposition of collagen rich extra-cellular matrix. The activated hepatic stellate cells (HSCs) are primary source of liver fibrotic tissue. In spite of intensive research, the mechanisms of fibrosis and activation of HSCs is not completely understood and the development of efficient anti-fibrotic agents remains a priority. Autophagy, an intracellular degradation pathway, plays an important role in diverse cellular processes in health and disease. Autophagy has several beneficial effects on the hepatocytes including insulin sensitivity and degradation of intracellular lipids in the hepatocytes (lipophagy). Autophagy also prevents hepatocyte injuries including those caused by oxidative stress and inflammatory cytokines. The role of autophagy in HSCs is, however, ambiguous. Numerous studies depict autophagy as a pro- fibrogenic process due to its association with HSC activation and liver fibrosis. The main mechanism underlying autophagy-mediated HSC activation and fibrosis is autophagy-mediated lipid droplet (LD) depletion which has been shown to concur with HSC activation. On the other hand, several studies show anti-fibrotic effects of autophagy via promotion of HSC death, degradation of pro-fibrotic mediators including collagen and metalloproteinases, and inhibition of pro-fibrotic signal carrying exosomes. These discrepancies may be attributable to the traditionally inefficient and inaccurate autophagy markers, instances of lack of cell-type specific studies, context of autophagy modulation in relation to the hepatic injury, and the role of autocrine/ paracrine signaling in fibrosis. It is noteworthy that several anti-fibrotic agents including statins are known to induce autophagy. We hypothesize that precise morphological detection of autophagic process will clarify the role of autophagy in HSC activation. We will address this hypothesis with the following specific aims: 1) To morphologically determine the status of autophagy during HSC activation using a novel HaloTag- LC3 assay; 2) To study the inter-relationship between lipid accumulation and autophagy in the HSC activation; and 3) To delineate the effect of ROS scavenging on lipophagy and lipid metabolism in HSC fibrogenic response. By employing the novel HaloTag-LC3 assay which can detect specific autophagy structures and high parameter metabolic flow cytometry (MetFlow), we will determine the state and role of autophagy in primary HSCs in the context of HSC activation, HSC-hepatocyte interaction, and lipid metabolic profile. Once completed, this study will provide a unique knowledge about the specific steps of autophagy involved in HSC activation and provide a foundation for mechanism-based therapeutic tools against liver fibrosis.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY Obesity is a global epidemic that is associated with sympathetic overactivation, endothelial dysfunction, and insulin resistance; all of which can contribute to hypertension development. Despite well-established clinical associations, the mechanisms connecting obesity with hypertension remain poorly understood, with several antihypertensive therapies eliciting detrimental metabolic effects. This illustrates the need to identify new targets with a positive metabolic profile for treatment of obesity hypertension. We propose that angiotensin-(1- 7), a protective hormone of the renin-angiotensin system, provides this ideal target. In support of this concept, we have shown that acute and chronic angiotensin-(1-7) treatment reduces cardiovascular sympathetic tone and blood pressure and improves endothelial function and insulin sensitivity in obese mouse models. There is limited clinical data, however, with no information on systemic effects of angiotensin-(1-7) on cardiovascular or metabolic outcomes in any patient population including obesity hypertension. To address this critical gap, we obtained preliminary data in obese hypertensive subjects suggesting that: circulating angiotensin-(1-7) levels are reduced and negatively correlate with blood pressure; acute intravenous angiotensin-(1-7) infusion reduces blood pressure and may increase endothelial-dependent vasodilation and decrease sympathetic tone; and angiotensin-(1-7) may improve insulin sensitivity, findings consistent with the positive metabolic effects seen in animal models. Based on these data, this proposal will test the central hypothesis that angiotensin-(1-7) improves cardiovascular and metabolic-related derangements in obesity hypertension. We propose proof-of- concept mechanistic studies to examine the cardiovascular and metabolic effects of acute intravenous angiotensin-(1-7) infusion in obese hypertensive subjects. We will test hypotheses that angiotensin-(1-7) can: reduce sympathetic activity and blood pressure; improve endothelial function; and increase whole-body insulin sensitivity and insulin-mediated microvascular recruitment. A sub-study will also be performed in pure autonomic failure patients lacking autonomic modulation of blood pressure to determine if angiotensin-(1-7) blood pressure and vasodilatory effects are via direct actions on the vasculature versus indirect engagement of autonomic mechanisms. Any results obtained from these studies will be novel and will increase mechanistic understanding of the role of angiotensin-(1-7) in the etiology of obesity hypertension.

NIH Research Projects · FY 2025 · 2024-04

Project Summary/Abstract Mitochondrial dysfunction is the main mechanism contributing to cell death and tissue damage in cerebral ischemia-reperfusion injury (I/R). Calcium-induced mitochondrial swelling, uncoupling of respiration, and cell death occur in mammals under hypoxic conditions of I/R. These changes are due to the activation of the mitochondrial megachannel also known as the mitochondrial permeability transition pore (mPTP). Remarkably, the embryos of the mysterious organism, brine shrimp Artemia franciscana, tolerate anoxic conditions for several years and were reported to lack hypoxia and Ca2+-activated mPTP opening. mPTP was at the center of scientific research for several decades and it still remains one of the most mysterious phenomena in biology today due to the lack of information about its exact molecular composition, structure, and regulatory mechanisms. Recently, mammalian ATP synthase was shown to undergo calcium-induced conformational changes to form an uncoupling channel with the biophysical properties of mPTP. In this proposal, we will use single-particle cryo- electron microscopy (cryo-EM) to identify the high-resolution structure and activation/inactivation mechanisms of Artemia franciscana ATP synthase. These studies will reveal the distinct structural features of Artemia ATP synthase that prevent the Ca2+ and hypoxia-induced mitochondrial dysfunction and cell death in this organism. Successful completion of this proposal will help to unlock the development of specific and potent therapeutic compounds targeting mammalian ATP synthase leak channel for treating I/R injury and other mPTP-related diseases.

NIH Research Projects · FY 2026 · 2024-04

ABSTRACT / PROJECT SUMMARY Background: Acute pancreatitis (AP) is a common and serious condition, accounting for more than 300,000 hospitalizations and over $2 billion in health care costs per year in the United States alone. Patients with AP have an increased risk of several complications including diabetes (DM). Two recent meta-analyses suggest that AP increases risk of DM by 23-37%, and that the risk increases over time. The pathogenesis of DM following AP has historically been ascribed to the loss of β-cell mass, however other mechanisms, including the development of insulin resistance or β-cell autoimmunity may also play a role. The Type 1 Diabetes and Acute Pancreatitis Consortium (T1DAPC): The overall goals of the T1DAPC are to describe the risk for and mechanisms of diabetes mellitus occurring after one or more episodes of acute pancreatitis. Secondary objectives include defining the natural history of acute pancreatitis, the clinical risk factors that predict development of diabetes and pre-diabetes, the natural history of beta cell function after acute pancreatitis, and the potential contribution of an altered immune state to the increased risk for diabetes after acute pancreatitis. These objectives are being addressed the ongoing, prospective, observational T1DAPC study entitled “Diabetes RElated to Acute Pancreatitis and its Mechanisms (DREAM)”. DREAM-ON: This ancillary study pilot proposal, entitled “Diabetes RElated to Acute Pancreatitis and its Mechanisms: Metabolic Outcomes Using Novel CGM Metrics (DREAM-ON)” which will be conducted in participants enrolled in the DREAM study, will test whether continuous glucose monitoring (CGM) is clinically useful to predict risk for developing diabetes, the need for insulin therapy among those who develop diabetes and whether CGM can provide insight into the diabetes subtype among patients who have experienced an episode of acute pancreatitis. Thus, the results of this ancillary study will complement and extend the primary research questions in the DREAM study. Specific Aims: Aim 1: To perform CGM in all participants in the DREAM study in conjunction with their scheduled visits at months 3, 12, 24 and subsequent annual visits and to determine if CGM metrics predict incident pre-diabetes and DM. Aim 2: To determine if CGM metrics predict need for insulin therapy in patients who develop DM after AP. Aim 3: To determine whether CGM metrics correlate with measures of insulin secretion and insulin resistance derived from the OGTT, MMT and FSIVGTT, and can be used as a surrogate to predict diabetes subtype.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY Improved treatment of colorectal cancer (CRC) requires better understanding of key regulators of survival and proliferation of intestinal epithelial cells and their crosstalk with immune system. Upregulation and hyperactivation of the β-TrCP E3 ubiquitin ligase underlie CRC pathogenesis and poor responses to chemotherapy. While directly inhibiting β-TrCP proved to be practically difficult, here we propose to target the key regulators of β-TrCP in CRC including proteins that stabilize its mRNA (IGF2BP1) and protein (PARP11). Our published and preliminary data indicate that IGF2BP1 and PARP11 act within malignant cells as well as within benign immune cells of the tumor microenvironment (TME) to stimulate colorectal tumors growth and progression and limit their responses to chemotherapeutic regimens. These data support an overarching hypothesis that targeting IGF2BP1 and PARP11 should elicit potent anti-tumor effects and improve the efficacy of anti-cancer chemotherapies. We will test this hypothesis in three independent yet integrated Specific Aims. In Aim 1 we will determine the roles of IGF2BP1 in cancer cells and in the pro-tumorigenic TME and in responses to therapy. We will utilize genetic modulation of IGF2BP1 expression in CRC cells and cells of the TME to understand the role of IGF2BP1 for tumor growth, metastases, and sensitivity to the FOLFOX regimen, which is a standard of care chemotherapy for CRC patients. Mechanisms governing IGF2BP1 expression in TME as well as mechanisms of TME-dependent involvement of IGF2BP1 in sensitivity to chemotherapy will be investigated. Aim 2 will determine the importance of PARP11 in growth and progression of intestinal tumors as well as their responses to anti-tumor therapy. We will use genetic, molecular, and pharmacologic approaches to modulate PARP11 in colon adenocarcinoma cells or in the non-malignant TME cells to determine the importance of PARP11 in pro-tumorigenic phenotypes in vitro and ability to form orthotopic tumors and liver metastases in vivo. PARP11-dependent mechanisms underlying control of malignant cells by anti-tumor immune system and responsiveness of tumors to chemotherapy will be investigated. Aim 3 will examine the effects of pharmacological targeting of the regulators of β-TrCP to improve the responses of colon adenocarcinoma tumors to chemotherapy. We will determine the anti-tumor effects of novel, potent and highly selective inhibitors of IGF2BP1 (AVJ16) and PARP11 (ITK7) as mono-agents and their ability to improve the responses to FOLFOX regimen. Completion of these studies should gain the insight on the importance of IGF2BP1 and PARP11 in colorectal tumor growth and metastasis, and characterize these regulators as potential vulnerabilities, which could be exploited for improving chemotherapy of CRC.

NIH Research Projects · FY 2026 · 2024-02

Project Summary Personality disorders are complex and debilitating disorders associated with profound risk for suicide and substantial cost related to long-term intensive care, emergency room visits, and psychiatric hospitalizations. Although traditionally diagnosed only in adults, emerging risk for personality pathology can be reliably assessed in early adolescence. The level of personality functioning (LPF) is a dimensional measure of personality pathology that predicts the development of personality disorders across the lifespan and contributes to long-term dysfunction beyond the effects of other clinical concerns, such as anxiety, depression, attention- deficit/hyperactivity disorder, and conduct problems. LPF refers to the degree of disturbance in self (i.e., identity) and other (i.e., interpersonal) functioning, and as a dimensional measure, is sensitive to the individual differences in emerging identity and interpersonal development in early adolescence. Yet very little research has examined neurophysiological mechanisms of LPF in youth, missing critical early opportunities to target youth at risk for some of the most stigmatized and costly conditions. Deficits in the NIMH RDoC constructs of Attachment and Affiliation and Understanding of Self may underlie impaired LPF. At the self-report level, impaired LPF is associated with difficulty modulating responses to social acceptance cues and accurately encoding self-relevant information. However, examining these processes at the neural level, which is not confounded by lack of insight or other demand characteristics, may more clearly elucidate processes underlying impaired LPF. Individual differences in processing social acceptance cues and self-referential information can be reliably detected at the neural level using event-related potentials (ERPs), but have not been applied to understanding personality disorders in youth. For this work, a sample of 240 early adolescent youth (ages 10-13, 50% female) will be recruited, the majority (n=180) from two clinically diverse outpatient psychiatry clinics, while the remaining (n=80) youth will be comparison youth without a history of psychopathology. Neurophysiological assessments of social acceptance processing and self-referential processing will be administered, and will be tested as predictors of growth in LPF impairment measured over two years. LPF will be assessed using behavioral observations of parent and peer interactions and through daily ecological momentary assessment (EMA) collected over two years. Additionally, the effect of LPF on clinical indicators, including functional impairment, suicidality, and treatment use, will be tested, considering other key clinical concerns, including maladaptive traits, co-occurring mental health disorders, substance use, and sociodemographic and developmental factors. Results from this work will provide evidence to elucidate treatment targets to mitigate the long-term personal and societal burden associated with personality disorders.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY Age-related macular degeneration (AMD) is a leading cause of vision loss, yet the molecular events that initiate the early retinal defects that lead to visual dysfunction remain poorly understood. The objective here is to examine a role for the stress response protein regulated in development and DNA damage 1 (REDD1) in the impaired adaptive response of retinal pigment epithelium (RPE) with aging and consequently development of AMD. Proof-of-concept studies support a potential role for REDD1 in AMD pathophysiology. In fact, intravitreal administration of a siRNA targeting the REDD1 mRNA has demonstrated promise for improving visual function in patients with AMD. However, we recently discovered that the REDD1 protein acts as a molecular redox sensor. Specifically, a reversible redox-sensitive disulfide bond prevents degradation of REDD1 by lysosomal proteolysis. Consequently, retinal REDD1 inhibition by siRNA-mediated knockdown may only be partially effective for reducing REDD1 protein expression in retinal disease. The central hypothesis is that activation of the REDD1 redox-sensor in RPE promotes oxidative stress, inflammation, and retinal pathology in AMD. To test the hypothesis, we will employ a preclinical murine model of non-neovascular AMD, transgenic REDD1 mouse lines, and human iPSC-derived RPE in three specific aims. Aim 1 will examine a role for REDD1 as a dominant governor of the nuclear factor erythroid-2-related factor 2 (Nrf2) antioxidant response in RPE. We predict that REDD1 prevents a proper antioxidant response in AMD by promoting Nrf2 nuclear exclusion. Aim 2 will investigate a role for REDD1 in RPE inflammation and trans-differentiation via activation of the transcription factor nuclear factor kappa B (NF-κB). The proposed studies will explore NF-κB signaling in AMD models and evaluate the expression of NF-κB target genes, including pro-inflammatory cytokines and epithelial-to- mesenchymal (EMT) transcription factors. Aim 3 will utilize REDD1 knockout mice and a newly developed REDD1 point mutation knockin mouse that expresses a REDD1 variant that is continuously degraded by lysosomal proteolysis to evaluate the formation of hyper-reflective foci, RPE damage, photoreceptor thinning, and impaired visual function in non-neovascular AMD. It is well established that oxidative stress and inflammation are crucial factors in the development and progression of the complications that cause visual impairment. The studies herein are significant because they are designed to identify and characterize specific molecular events that contribute to oxidative stress and inflammation in AMD by addressing key knowledge gaps related to a cutting-edge therapeutic target. To do so, we will explore the concept that activation of the REDD1 redox sensor is a unifying molecular mechanism for improper activation of Nrf2 and NF-κB in AMD.