Temple Univ Of The Commonwealth

NIH Research Projects · FY 2025 · 2024-03

Anorexia nervosa (AN) is a deadly mental health disorder characterized by pronounced low bodyweight resulting from reduced food intake and oftentimes simultaneous hyperactivity, typically through compulsive exercise. Individuals experiencing AN, who are at least 75% more likely to identify as female, bear the brunt of this condition. AN has the second highest mortality rate among mental health disorders, recently surpassed only by opioid-use disorder. In addition to the strikingly low bodyweight, AN leads to chronic medical conditions and lifetime disability. The overwhelming majority of deaths in AN patients are attributed to either unknown medical complications or suicide. Interestingly, the prioritization of hyperactivity over food intake, a prominent feature in many individuals with AN, has been observed not only in humans but also in various species, including rodents. Leveraging this parallel, researchers have employed the activity-based anorexia (ABA) rodent model to shed light on the evolutionarily conserved motivational drives potentially underlying AN. Nevertheless, the specific neurobiological factors driving these maladaptive behaviors in ABA and AN are still largely unknown. Our previous studies have implicated hypothalamic hunger-promoting agouti-related peptide (AgRP) cells as potential contributors to the valence reassignment that occurs in AN between hunger, feeding, and hyperactivity. Notably, we have demonstrated that increasing AgRP neural activity can promote bodyweight gain in ABA mice, offering a potential therapeutic target for reversing weight loss in ABA and, potentially, AN. However, the precise mechanisms through which AgRP neurons coordinate these effects and whether they modulate valence associations with hunger, physical activity and feeding behavior in ABA remain elusive. The objective of this proposal is to determine the neurobiological mechanisms employed by AgRP neurons to orchestrate the re- prioritization of feeding behavior over hyperactivity in ABA, utilizing cutting-edge behavioral technologies and knock-in mouse models. In Aim 1, we will expand upon our previous findings to determine if AgRP cells can sense and modulate both hyperactivity and feeding during ABA. Through in vivo fiber photometry and optogenetics, we monitor and manipulate AgRP populations, respectively, in ABA and control mice while they are freely behaving on the behavioral paradigm. In Aim 2, we will unravel the circuits involved in AgRP-mediated bodyweight gain in ABA mice using projection-defined chemogenetics. By defining and characterizing the cellular and circuit functions of AgRP neurons in female ABA mice, our research endeavors hold the potential to identify a therapeutic target for the re-prioritization of feeding over exercise in AN. Ultimately, our findings have the potential to alleviate the chronic disability associated with AN, leading to improved survival and enhanced quality of life for individuals affected by this severe disorder.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY The relationship between alcohol use (AU) and aggression (AGG) is widely recognized. In fact, epidemiologic data demonstrate that 40% of individuals with Alcohol Use Disorder (AUD) in the community display clinically significant levels of AGG. Moreover, aggression frequency in individuals with AUD+AGG increases as overall alcohol consumption increases, making alcohol- facilitated aggression a major public health threat. While the relationship between AUD and AGG may be bidirectional, our data indicate that more than 90% of those with AUD+AGG report the onset of problematic aggression before the onset of problematic drinking. As such, premorbid AGG represents a risk factor for the development and maintenance of problematic drinking. In addition, experimental studies suggest that alcohol increases aggression primarily in those who are already aggressive. To date, the literature contains little treatment research in those with AUD+AGG. More importantly, AUD treatments, in general, minimally focus on treating AGG in AUD, including considering it an optional element of care. Fortunately, full cognitive behavioral treatments for AGG exist, notably CRCST (Cognitive Restructuring, Relaxation and Coping Skills Training) which we have shown has good efficacy in reducing aggression in those with AGG. In addition, based on our work in aggression, we posit that individuals with AUD+AGG constitute an important subgroup of those with AUD and that this comorbidity (AUD+AGG) represents a target for a personalized medicine approach for AUD. At this time, we have developed a provisional Integrated CBT manual containing the most critical elements for treating AUD and for treating AGG. In this R34 project, we will refine our Integrated CBT-AUD+AGG Treatment Manual in a small study in patients with AUD+AGG and then conduct a pilot RCT comparing this integrated CBT AUD+AGG intervention with CBT-AUD Alone (from Project MATCH). The goal of this study is to obtain acceptability, feasibility, and effect size data, which will be used inform the next steps in this program of research (e.g., a large scale clinical trial).

NIH Research Projects · FY 2026 · 2024-02

Project Summary Acute myeloid leukemia (AML) is the most common diagnosed adult leukemia, the median age of patients with AML is about 70 years. Although the prognosis for younger adults with AML has improved during the last four decades, there has been little progress in the treatment of older adults. Currently, approximately 90% of adults with AML over the age of 55 will die due to resistance to therapy, relapse, or complications from harsh treatments such as chemotherapy. AML disease progression is heavily influenced by supportive cells in the tumor microenvironment. Bone marrow mesenchymal stromal cells (BMSCs) are an instrumental extrinsic component to normal hematopoiesis which are hijacked by leukemic cells in the process of leukemia development. Based on AML being mainly a disease of older adults and evidence of an accelerated aging phenotype in the (BM) microenvironment of AML, this proposal aims to investigate the role of aging and senescence in AML disease progression and to ultimately identify therapeutic targets and eliminate the leukemia-supportive aging phenotype in the BM. Although epigenetic aging and senescence are two distinct but parallel mechanisms of aging, they have been shown to converge where certain triggers of senescence can affect epigenetic age. The molecular basis for age-related alterations in AML-derived BMSCs are poorly described and if deciphered, could have significant implication on both the prevention and treatment of elderly AML. Moreover, the correlation of epigenetic age in cells of the AML tumor microenvironment with outcome has not been examined. Thus, the specific aims of this proposal are to (1) examine epigenetic, transcriptional and phenotypic differences in BMSCs derived from AML patients, compared to age matched control BSMCs, enabled by the use of methylation studies, sequencing, mass cytometry and biochemical assays (2) determine the epigenetic age via methylation analysis of different components of the tumor microenvironment (T-cells, tumor cells and BMSC cells) in AML patient samples and correlate with disease outcome and finally, (3) utilize findings and techniques developed in aim 1 and 2 to study the status of epigenetic aging and senescence in in vitro and in vivo models of accelerated aging and relapse to determine if they can be therapeutically targeted. The completion of this work will potentially provide a quantitative measure of senescence in elderly AML patients, further enhance risk stratification, and will help identify novel age-related targets in AML-BMSC with potential to lead to development of new therapies.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY/ABSTRACT Heat waves are a growing public concern and a leading cause of climate-related morbidity and mortality in the US. Eight out of 10 U.S. population live in urban areas. The combination of rising temperatures and urbanization poses a significant risk for vulnerable urban populations due to the slow release of heat by urban heat islands created by extreme summer temperatures. While adaptation planning for heat waves during the day has been developed and implemented at the city-wide and governmental levels, heat adaptation behaviors in individual households are less understood, particularly during nighttime hours. Although prior studies have identified several individual factors associated with heat adaptation, the role of nighttime indoor heat exposure as an external hazard has not been examined. Moreover, the contribution of the built environment on heat exposure and individual adaptation behavior has not been fully evaluated although the build environment play a role in residential household. To fill these gaps, we will examine the combined effects of individual factors, built environment, and nighttime indoor heat exposure on heat adaptation behavior using the Protective Action Decision Model. The long-term goal of this study is to reduce heat exposure risks and increase resilience to climate change for vulnerable urban populations. Our aims are to establish a community advisory board and examine the effects of indoor home heat exposure, individual attribute, and built environmental factors on heat adaptation behavior. The study results will be disseminated to community leaders and residents. We expect that disseminating our findings regarding the drivers and barriers of heat adaptation behavior helps individuals living in underserved urban communities take the necessary steps to reduce their potential health risks associated with nighttime heat exposure. We believe that the results of this study can provide an affordable and practical adaptation strategies for those affected by heat exposure during nighttime. This proposed exploratory study is directly responsive to the National Institute of Environmental Health Science Strategic Goal 2: Promoting Translation- Data to Knowledge to Action: Emerging Environmental Health Issues (Climate Impacts on Health and Preparation for Future Threats) and has the potential to reduce heat exposure risks and increase resilience to climate change for vulnerable urban populations.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY Oral squamous cell carcinoma (OSCC) exerts a significant clinical and financial burden worldwide. Recently, there has been increasing interest in the role of the microbiome in OSCC. Among microbial species that have frequently been identified in association with OSCC and demonstrated to promote oral carcinogenesis, both in vitro and in animal models, include the bacterium Fusobacterium nucleatum and the fungus Candida albicans. The two species have been demonstrated to interact via co-aggregation; however, whether such interkingdom interaction can promote oral carcinogenesis has never been explored The current proposal builds on our previous studies investigating the microbiome associated with OSCC and oral leukoplakia in clinical samples, and assessing the effects of oral bacteria against oral epithelial cell lines in vitro. The proposed studies will investigate for the first time the potentially synergistic interaction between C. albicans and F. nucleatum in malignant progression, which we hypothesize is facilitated by their coaggregation. Based on our preliminary data, we also hypothesize that the two species mediate part of their oncogenic properties through upregulation of INHBA, a proposed oncogene acting through the TGF-β pathway. To address these hypotheses, we propose to assess synergistic effects of C. albicans and F. nucleatum on normal, dysplastic, and neoplastic oral epithelium in vitro (Aim 1), and to study the carcinogenicity of C. albicans and F. nucleatum co-carriage in 4- nitroquinoline-1-oxide-induced OSCC mouse model (Aim 2). Combinations of wild-type, aggregation +ve strains and mutant, aggregation-deficient strains of the two species will be used in the two aims to assess the role of co-aggregation in promoting synergistic carcinogenicity. The involvement of INHBA upregulation in this synergy will be investigated by mechanistic gene knockdown experiments. The project will employ a range of technologies including cellular and biochemical assays, metatranscriptomics, histopathology, immunohistochemistry, flow cytometry, q-PCR, fluorescent in-situ hybridization and 16S sequencing to investigate the effect of treating the cell lines and mice with the test species. This innovative, exploratory study leverages the complementary expertise of the research team to provide a first insight into the potential role of interkingdom microbial interactions in OSCC and shed light on novel mechanisms by which C. albicans and F. nucleatum may contribute to oral carcinogenesis

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY/ABSTRACT Individuals with schizophrenia spectrum disorders experience a range of symptoms which cause high levels of functional impairment, thus representing a large personal and public health burden. Similarly, individuals at clinical high risk (CHR) for the development of psychosis experience general work and academic impairments, social impairment, and reductions in quality of life that are evident even prior to the onset of full-threshold psychosis. Therefore, studies evaluating CHR populations can help to uncover how symptoms of schizophrenia first arise and indicate more effective avenues for early intervention. Of particular importance is the examination of negative symptoms, such as anhedonia, as they tend to present prior to the onset of positive symptoms, are more resistant to treatment, and are more strongly predictive of functional outcomes. Further, although anhedonia commonly occurs in those with psychosis, it also is a hallmark feature of depression, a disorder with high comorbidity in CHR individuals. Mounting evidence suggests that childhood trauma not only increases risk of developing psychosis, but is associated with anhedonia, specifically, across diagnoses, including for individuals with depression, as well as non-psychiatric controls. Further, childhood trauma has deleterious effects on the mesolimbic system, particularly the hippocampus, a region which animal models have recently implicated in the development of reward deficits underlying anhedonia. Despite these converging lines of research, no one, to date, has examine the mediating role of mesolimbic functioning in the relationship between childhood trauma and anhedonia. Therefore, the proposed study will examine the association between childhood trauma and anhedonia via the indirect effect of hippocampal function during novelty processing, which has been shown to engage dopaminergic reward systems. The proposed study, in the context of my sponsor's NIMH-funded R01, will recruit non-help-seeking 16-30 year olds who represent the full spectrum of psychosis-risk to complete self- report questionnaire and clinical interviews, with a subset completing both resting-state and task-based functional magnetic resonance imaging (fMRI) scans. Findings from the proposed study have the capacity to provide additional insight into previously established reward-related alterations associated with anhedonia, thus providing a novel framework to understand the relationship between childhood trauma and anhedonia. Further, we will explore potential differences in the association between childhood trauma, mesolimbic engagement, and anhedonia across CHR and non-CHR individuals to assess specificity. To complete this study, a training plan has been developed that consists of formal coursework, workshops, experiential learning, and mentorship. This fellowship would allow me to obtain additional training opportunities that would not otherwise be available to me, in order to develop the expertise in the pathophysiology of reward dysfunction, neuroimaging, and data analysis necessary to become an independent clinical scientist.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY/ABSTRACT Symptoms of social anxiety rise dramatically at the transition to adolescence (ages 10-11), as peers increase in importance. Failure to remit in social anxiety by mid-adolescence (ages 12-13) predicts increased symptom severity, comorbid psychopathology, and diminished quality of life in adulthood. It is therefore critical to identify factors that contribute to remittance or persistence of social anxiety during the transition to adolescence, as such factors may serve as treatment targets. While much prior research on social anxiety persistence focuses on threat-based processes, gold standard treatments that target these processes rarely result in remission. Thus, there is an urgent need to develop a novel framework of investigation to identify new pathways to persistence and corresponding intervention targets. While prominent theories suggest that biased memory for negative social outcomes contribute to symptoms, such biases are relatively understudied in adolescent social anxiety. This omission is significant because memory is reconstructive and fundamentally malleable, making it an ideal target for treatment. Our preliminary data using behavior, fMRI, and neuromelanin-sensitive MRI (NM-MRI) to quantify dopamine function, suggests interplay between dopamine system function and medial prefrontal cortex (mPFC)- striatal connectivity promotes memory bias in adolescent social anxiety. Our overall goal is to apply this multi- systems approach to the longitudinal study of adolescent social anxiety. Doing so may help identify sorely needed biobehavioral targets for intervention during a key period of development, thereby preventing devastating lifelong problems. To achieve this, we will assess social anxiety symptoms, dopamine system function and brain- based functional connectivity associated with memory bias in 255 youth (10-11 years). Additional assessments of symptom expression obtained at 6-month intervals as they transition to mid-adolescence (12-13 years) will enable trajectory mapping. Our central hypothesis is that in the presence of high dopamine system function, youth with biased recall for negative peer feedback and enhanced mPFC-striatal connectivity will be more likely to persist in social anxiety versus those who remit. Specific aims are to (1) characterize longitudinal associations between memory bias and social anxiety during a critical developmental period; (2) determine the extent to which memory bias mediates the relation between mPFC-striatal connectivity and social anxiety symptom trajectories; and (3) test moderation of brain-memory bias relations by dopamine system function. This proposal will advance the field and provide novel treatment targets by characterizing the contribution of memory bias to adolescent social anxiety persistence and remittance and its neural mechanisms. Innovative features are a focus on understudied mechanisms (memory) in a key age (10-13 years) using multiple imaging modalities including NM- MRI. Our project is significant because it is a critical first step towards providing the right treatment (based on neural mechanisms) to the right youth (those who persist in social anxiety), at the right time (early adolescence, before social anxiety becomes intractable).

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY: Updates: I am moving to a new position at Temple University Lewis Katz School of Medicine as an independent, tenure-track Assistant Professor in the Department of Neural Science. Here, I will conduct the R00 phase experiments and develop my own independent research program. The overall aims of the R00 phase have not changed from the K99 proposal. The brain has two strategies for controlling behavior. Goal-directed actions that rely on prospective consideration of their outcomes and consequences, and habits, reflexive responses performed without forethought of their consequences. Overreliance on habit can lead to maladaptive perseverative behavior, which contributes to numerous psychiatric conditions, including autism spectrum disorders. Environmental factors, like stress, and genetic alteration, can tip the balance between actions and habits. However, our knowledge of the mechanisms by which these factors influence habits is lacking, limiting our understanding of maladaptive behaviors in psychiatric conditions, and how to treat them. Therefore, the broad goal of my research is to reveal specific neuronal mechanisms that allow gene x environment interactions to disrupt behavioral strategy. Accumulating evidence suggests the dorsomedial striatum (DMS) is central for controlling goal-directed actions. Inhibition of the DMS, particularly D1+ neurons, disrupts goal-directed control, resulting in a bias toward habits. The basolateral amygdala (BLA) is a known hub for stress responsivity in the brain and projects onto DMS Drd1+ neurons. My postdoctoral work has revealed that BLA-DMS activity supports goal-directed learning and its suppression is necessary for premature habit formation. Further, I found that activation of BLA-DMS projections in stressed animals is sufficient to restore goal-directed control. This led me to the intriguing hypothesis that chronic stress dysregulates DMS D1+ control of goal-directed learning via BLA-DMS input. In my K99 phase, I found that indeed, suppression of DMS D1+ activity is necessary for chronic stress to promote premature habits. In my independent phase, I will apply these findings to a mouse model of 16p11.2 microdeletion, a genetic alteration associated with autism and that is known to dysregulate striatal function. Specifically, I will examine the vulnerability to premature habits of 16p11.2 mice, normally and after sub-threshold stress, and assess amygdala-striatal control, using cell-type specific in vivo calcium imaging with miniscopes, slice electrophysiology, and projection-specific chemogenetic manipulations. My findings will provide a mechanistic understanding of habitual control normally, after stress, and in a model of autism-associated genetic alteration. This will facilitate future work into the molecular and cellular mechanisms of this phenomenon and ultimately serve my goal of improving treatment approaches for psychiatric conditions.

NIH Research Projects · FY 2025 · 2023-11

PROJECT SUMMARY/ABSTRACT Childhood maltreatment impacts over a third of children worldwide and contributes to a quarter of all psychiatric disorders. Because parents with, compared to without exposure to childhood maltreatment are 2-3 times more likely to abuse their own children, many families are trapped in intergenerational cycles of abuse. Although intergenerational abuse is well-established, the cognitive mechanisms that contribute to its persistence are poorly mapped. We propose that childhood abuse may bias individuals towards prioritizing the encoding and recall of negative, rather than positive, social feedback. Despite robust evidence that childhood abuse is associated with negative cognitive biases as well as alterations in episodic memory and corresponding hippocampal dysfunction, the role that memory bias plays in intergenerational abuse remains untested. Because memory is reconstructive, it is fundamentally malleable, thus neurocognitive mechanisms of memory bias may serve as an ideal intervention target. One factor contributing to a dearth of research testing effects of memory bias on abuse transmission is a lack of ecologically-valid paradigms that link brain response to encoding social feedback with its subsequent recall. We addressed this challenge with the Recall After Feedback Task (RAFT). Preliminary work with the RAFT demonstrates that more severe childhood abuse is associated with greater recall for negative social feedback and enhanced anterior hippocampal engagement when encoding negative vs. positive social feedback. I propose to extend this work in parents with a range of childhood abuse experiences and test the extent to which neurocognitive mechanisms implicated in social memory bias relate to current harsh parenting style. This inquiry is essential given that abuse is an inherently social process, yet most studies of intergenerational transmission fail to probe neurocognitive mechanisms that support social processing. I aim to characterize associations between childhood abuse and 1) memory bias for social feedback; 2) the relationship between hippocampal activation when encoding social feedback and memory bias; and 3) the degree to which memory bias and brain activation promote harsh parenting. The proposed study will advance the field and provide novel intervention targets by characterizing the contribution of social memory bias and its neural basis to the link between childhood abuse and harsh parenting. The proposed training plan, which consists of workshops, experiential learning, and mentorship, will develop my expertise in ecologically-valid fMRI study design and analysis, neural mechanisms of memory, and neuropsychosocial outcomes following maltreatment. As a result, I will gain expertise and preliminary data needed for future grants that inform interventions to curtail intergenerational abuse and establish the foundation for becoming an independent developmental social neuroscientist. Temple University’s Department of Psychology and Neuroscience has a successful track record of conducting impactful NICHD-funded work and is the ideal setting for the proposed research and training.

NIH Research Projects · FY 2024 · 2023-09

Abstract Although exposure to microplastics and nanoplasctis (hereafter, microplastics) in general populations has been widely assessed, workers’ exposures to microplastics at workplaces are mainly unknown. Limited laboratory studies estimated that workers are exposed to airborne microplastics several orders of magnitude higher than general populations. Moreover, inhaled microplastics were found in all parts of the lung in humans and found significantly higher levels in the lower lung. Thus, workers handling or manufacturing materials containing microplastics are at increased risk for adverse health effects (e.g., respiratory outcomes) from microplastic exposures at their workplaces. To our knowledge, no studies have directly examined personal exposure to microplastics among workers, especially in the Service Sectors. Textiles made from synthetic (plastic) fibers are significant sources of occupational exposure to microplastics. Although workers in the drying cleaning and laundry industry are exposed to synthetic fibers (containing microplastics) by handling clothing and linens daily, these workers’ exposure to microplastics is largely unknown. Further, review studies summarized that workers in the garment industry handling synthetic fibers (including nylon, polyester, acrylic) had a higher prevalence of respiratory diseases than non-exposed populations. Hence, workers in the drying cleaning and laundry industry experience may experience a large burden of exposure to microplastics at their workplaces. In 2021, the dry cleaning and laundry industry in the United States employed 160,000 -244,000 persons, excluding self-employed persons. This industry is expected to grow nationally from $10.8 billion in 2020 to $14.4 billion by 2025. However, the impact of exposure to emerging microplastics on workers in this business sector is unknown due to the lack of exposure assessment. This proposed R21 study, in response to the National Occupational Research Agenda: the Services Sector and the Cross-sector of Healthy Work Design, will evaluate the effects of key determinants (work hazards, characteristics of workplace and workforce) on inhalation exposure to microplastics in the laundry and dry cleaning industry. We aim to (1) measure inhalation exposure to microplastics from 30 workers in the laundry and dry cleaning industry, and (2) determine the effects of work-related factors (workforce, workplace, and work hazards) on inhalation exposure to microplastics among these workers. The results will inform a future intervention study and training program to develop strategies for reducing workers’ exposure to microplastics in the Service Sector. The proposed research will also improve a scientific knowledge gap by informing workers’ exposure to microplastics that can be deposited in the human lungs, and by examining the association between microplastic exposure and adverse health effects among workers in the Services Sector.

NIH Research Projects · FY 2025 · 2023-09

The kappa opioid receptor (KOR) is one of the three opioid receptors. KOR agonists produce analgesic and anti-pruritic effects, but their development for clinical use has been limited by side effects, most importantly dysphoria and psychotomimesis. KOR antagonists display antidepressant- and anti-anxiety-like effects in rodents and may be useful for the treatment of drug addiction in humans. The paraventricular nucleus of the thalamus (PVT), the most dorsal nucleus of the thalamic midline nuclei, is among the brain regions that express high levels of KOR. The PVT receives inputs from the prelimbic, infralimbic and insular cortices, the ventral subiculum and many hypothalamic and brain stem nuclei. The PVT sends dense projections to several limbic structures including the amygdala, the bed nucleus of the stria terminalis, and the core and shell of the nucleus accumbens. The PVT is part of the brain anxiety network and is involved in stress responses, fear, anxiety, arousal, reward, and homeostasis. The KOR level in the PVT is similar to that in the ventral tegmental area, but KOR in the PVT has not yet been characterized. For the proposed studies, we have generated a mutant mouse line expressing tamoxifen-inducible Cre conjugated to KOR (KOR-iCre). In this application, we propose the following three specific aims. For the Aim 1, the origins of afferent projections to PVT KOR- expressing neurons will be determined by Cre-dependent rabies virus-mediated monosynaptic retrograde tracing. The brain regions of efferent projections of PVT KOR-expressing neurons will then be characterized using Cre-dependent anterograde tracing. Whether KOR+ neurons in the PVT projecting to different brain regions receive innervations from different brain areas will also be investigated. Finally, the origin of dynorphin inputs into the PVT will be explored. For the Aim 2, we will elucidate the functions of PVT KOR by examining the effects of conditional deletion of PVT KOR on behaviors such as KOR agonist-induced analgesia in a visceral pain model, conditioned place aversion, naloxone-precipitated withdrawal signs, aversion after chronic morphine and anxiety-like behaviors. For the Aim 3, we will examine the roles of PVT KOR-expressing neurons in stress-related behaviors, fear conditioning and aversion by activation and inhibition of these neurons via chemogenetic approaches. This will be the first time that these PVT KOR-expressing neurons and KOR per se are investigated in a comprehensive manner. Determining the neuronal circuitries in which the KOR is involved and the functional significance of KOR+ circuits will enhance our understanding of KOR functional neuroanatomy and KOR-mediated aversion, anxiety, stress responses and other psychopathology. The knowledge acquired may provide the neuronal basis for developing KOR antagonists as anti-anxiety agents.

NIH Research Projects · FY 2025 · 2023-09

Development of blood-based methylation biomarkers for CRC risk prediction Colorectal cancer (CRC) incidence and mortality rates are disproportionately higher in African Americans (AA) compared to Caucasian Americans (CA). Current non-invasive screening tools like fecal occult blood test (FOBT) or Cologuard detect cancer after it occurs, and more effective tools for prevention and treatment of higher risk individuals, such as colonoscopy or endoscopy, are invasive, less popular and subjective. Therefore, identification of early biomarkers that distinguish normal colon mucosa of cancer patients from normal colon mucosa of patients without cancer might decrease racial disparities in CRC. We have identified a subgroup of patients as having “Outlier Methylation Phenotype” (OMP) using normal tissue methylome. OMPs are highly epigenetically disrupted and display abnormal DNA methylation patterns throughout their epigenome. We have been able to significantly associate this phenotype with CRC patients over healthy controls. Furthermore, AA CRC patients appear more than twice as likely to have OMP than CA. In our current grant application, we propose to develop OMP as a less- invasive CRC screening and prognostic tool by evaluating the consistency of OMP status in a less-invasive (whole blood) tissue with an invasive tissue (normal colorectal mucosa). In Specific Aim 1A, we will test whether OMPs identified in colorectal tissues can also be identified in whole blood samples in 200 CRC patients (100 AA, 100CA) and age, sex, racial ancestry and location (for colorectal tissues) matched 400 healthy controls (200 with history of adenomas and 200 without history of adenomas) using epigenome-wide data from >850K CpGs. In Specific Aim 1B, we will analyze the association of OMP in CRC patients with known CRC molecular subtypes or mutations like CpG island methylation phenotype (CIMP), KRAS, BRAF, MLH1 to estimate whether or not OMP is a surrogate marker of any known CRC molecular subtype. In Specific Aim 1C, we will follow-up the controls (especially OMPs) after 3-4years of screening colonoscopy and compare the clinical outcomes to evaluate the relevance of this phenotype in screening or CRC prevention. We will also study the association of genetic (Specific Aim 2) and environmental (Specific Aim 3) factors with OMPs. In Specific Aim 2, we will genotype the blood DNA to confirm the self-identified racial ancestry of our samples and to study the association of genetic variants with abnormal methylation in OMPs. In Specific Aim 3, we will evaluate the effect of diet and social determinants of health on OMP. Overall, the proposed study aims to identify and characterize molecular markers (OMP) in a less- invasive tissue (whole blood) that can be used both as a diagnostic and a prognostic tool, especially in the underprivileged AA population who have the lowest CRC screening rates, highest CRC incidence and highest CRC mortality rates.

NIH Research Projects · FY 2024 · 2023-09

Project Summary Uncertainty is an often pervasive, stressful experience that arises when making judgments about others' beliefs, intentions, or emotions (i.e., ambiguous social situations). Excessive uncertainty can have pernicious effects upon memory, mood, and physical and mental outcomes. Yet, we understand little of how judgments of social certainty form over time, the neural circuitry underlying these judgments, and how these judgments meaningfully differ from non-social uncertainty sources (e.g., calculations, perceptions). Uncertainty is featured in every developmental stage but adolescents and adults differ in their appraisals of and responses to ambiguity; both social and non-social. Traditional univariate neuroimaging analyses, which compare average magnitudes of activation across broad neural regions, are unresponsive to the subtle pattern differences that characterize complex social cognition. Novel multivariate techniques, such as intersubject correlations (ISC), applied to dynamic, feature-rich, and ecologically-valid sources of social ambiguity are crucial for understanding fundamental aspects of social cognition but have not yet been applied to answer these important questions. F99 Phase: My proposed pre-doctoral project uses a novel study design in which adult participants continuously rate their certainty of a given social (e.g., a character's innocence or guilt) and non-social (e.g., frame luminance) outcome while observing long-form narrative video stimuli (i.e., 45 min crime drama) during fMRI. This yields a continuous time course of concurrently-recorded neural and behavioral data which can be analyzed via ISC to determine the neural circuitry commonly implicated in uncertainty judgment formation among normative adult populations. Interdomain neural-behavioral synchrony can underscore how social and non-social certainty judgment formation mechanisms differ. Defining normative adult neural uncertainty responses provides a crucial comparison to vulnerable populations with pronounced uncertainty responses, including autism spectrum, anxiety, and mood disorders. Training during this phase includes instruction in computational methods relevant to the neuroscience of social decision-making and neurodevelopmental theory to prepare for the K00 Phase. K00 Phase: This phase extends my pre-doctoral research by applying the F99 Phase study design to adolescent populations. This approach could identify whether adults and adolescents are generally relying on distinct or similar neural circuitry to assess the same stimuli and inform subjective indicators of uncertainty (i.e., behavioral ratings). Adolescence is a period marked by intense attention towards social others. The presence of ambiguous social stressors during this period predicts susceptibility to and severity of anxiety and depression into adulthood. Thus, how social uncertainty is processed has notable implications for researchers working at any later developmental stage. The training plan in this phase of the proposal will focus on developing expertise in the developmental social neuroscience literature, computational methods that can be applied to developmental social neuroscience research, and mentorship skills to establish a diverse, inclusive independent research lab.

NIH Research Projects · FY 2025 · 2023-09

SUMMARY The Collaborative ICPRS clinically-oriented predoctoral/postdoctoral administered development current proposal titled “Sequential Modeling for Prediction of Periodontal Disea ses: An I ntra- Practice-Based Research Study ( ICPRS)” is developed in response to RFA-DE-23-012 . The will seek to develop various tasks in 4 component sections. In Component 1 , we seek to train 10 faculty from Temple University Kornberg School of Dentistry (TUKSoD) nd ten students in Applied Clinica l Research by offering a graduate certificate program by the College of Public Health (CPH) on facilitating skills in applied patient-oriented research a . The 15-credit certificate will focus . In Component 2, we aim to strengthen collaborations between TUKSoD, CPH and the Medical School for faculty and student training, and overall research support for the execution of Component 4 of this proposal. Our proposed project was collaboratively developed by individuals with backgrounds in Periodontology, General and Oral Epidemiology, Oral Microbiome, Health Informatics, Biostatistics and Public Health, and Physicians who will assist in extracting medical metrics relevant to management of periodontal diseases (PD) from real-time medical records in the Pennsylvania Health Share Exchange (PA HSX). In Component 3, we aim to expand clinically-oriented faculty participation in grant funded research activities, in the annual research day, and in national research meetings by increasing the number of faculty-student mentoring partnerships to conduct small-scale research projects. Finally, in Component 4, we will develop longitudinal predictive models of PD incidence and progression using machine learning approaches and microbiome/metagenomic sequencing strategies. This component has the following aims: Aim1 seeks to develop clinical decision tools that utilize matched medical-dental patient datasets from TUKSoD and the PA HSX EHR records. Aim1a will focus on developing a Medical Continuity of Care Record (CCR) that will provide patients' up-to-date medical health metrics that can be used in the provision of care at the dental school. Using dental patient's matched records, Aim1b will build an AI Empowered Prediction Model for PD. Aim2 will build on findings from Aim1b to assess the potential of the subgingival and salivary microbiomes as additional predictors of PD incidence and progression. Aim 2a will validate a dysbiosis index, applied to both subgingival plaque and saliva samples, as a predictor of PD in a cohort study, while Aim2b will identify potential high resolution, salivary/plaque metagenomic biomarkers of PD progression via a nested case-control study. The proposed application builds on prior research conducted at TUKSoD and is directed towards increasing participation of clinically-oriented faculty and pre/postdoctoral dental students in the school's research activities, coupled with training in Applied Clinical Research including health informatics, opening a new pathway to further research in dental education as well as promote faculty professional development, expand the research academic workforce, and provide opportunities to conduct student-mentored practice-based research.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT Intermittent Explosive Disorder (IED) is the only disorder in the DSM-5 for which the cardinal symptom is recurrent aggressive outbursts. IED is a common and multidetermined mental illness, which, if left untreated, has a chronic and debilitating course that often involves interpersonal, occupational, and legal consequences, as well as adverse physical health outcomes. IED also is associated with a number of cognitive biases related to socioemotional information processing. Over-attentiveness to threatening information (attention bias) and the tendency to attribute hostile intent to others (interpretation bias) are known correlates of aggression more broadly, and aggressive behavior in IED specifically. Additionally, hyperarousal of the autonomic nervous system (ANS) following provocation has been demonstrated in aggressive individuals; however, this phenomenon has not yet been investigated in IED, nor has it been linked with the biased cognitive processing seen in the disorder. Understanding the biological correlates of such cognitive biases is crucial to developing an integrated biopsychosocial framework for IED. Further, understanding the mechanisms of cognitive bias modification (CBM) could advance targeted interventions for IED that address both cognitive and physiological dysfunction. Leveraging an existing randomized controlled trial led by my primary sponsor, the present study proposes to examine the relationship between cognitive (attentional and interpretive) biases and physiological reactivity following provocation both before and after a CBM intervention among individuals diagnosed with IED. At pre- treatment, both IED (n = 48) and non-aggressive personality disorder controls (PD; n = 48) will complete computerized measures of attention and interpretation biases, along with an aggression-provocation task (TAP; Taylor Aggression Paradigm), during which psychophysiological data (i.e., heart rate, respiratory sinus arrhythmia and electrodermal activity) will be collected. Following the pre-treatment assessment, participants in the IED group will be randomized into either (1) a four-week (8-session) computerized cognitive bias modification intervention (CBM; n = 24) or (2) a dose equated computerized placebo control (placebo, n = 24). In the CBM condition, the tasks are aimed at modifying the aforementioned attention and interpretation biases using corrective feedback between trials, whereas in the placebo condition, corrective feedback will not favor “non- hostile” responses, presumably leaving any existing cognitive biases unaffected. Post-treatment procedures will be identical to those at pre-treatment, in which cognitive biases are again measured via computerized tasks, and psychophysiological data are collected during the TAP. Results of this study will provide further insight into the pathophysiology of reactive aggression, the efficacy of CBM interventions for IED, and the development of novel interventions for IED that address cognitive and psychophysiological dysfunction that characterize the disorder. A training plan was designed to develop the applicant's expertise in the conceptual and methodological areas relevant to the proposed study and professional development toward becoming an independent researcher.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT The proposed study is designed to evaluate the efficacy of a remotely-delivered, acceptance-based behavioral intervention (ABTi) for individuals demonstrating weight regain after bariatric surgery by comparing it to a Control condition. The current study aims to: 1) evaluate ABTi's efficacy on weight outcomes compared to Control; 2) evaluate ABTi's efficacy on secondary outcomes including improvements in eating behaviors, physical activity, and medical conditions compared to Control; 3) test proposed mechanisms of action, including evaluating a) if changes in hypothesized active ingredients in ABTi mediate differences in weight outcomes between conditions and b) if the association between eating behavior and subjective internal states (i.e., hunger, cravings, urges, negative affect) will weaken in ABTi compared to Control. Patients 6 to 48 months out from surgery who have regained >5% of their weight will be randomized to ABTi or Control (n=200). Assessments willbe conducted at baseline, 3, 6 (end of active treatment), and 12 months (follow up) after enrollment. Developing innovative interventions to reverse postoperative weight regain is crucial to maintain the health benefits of surgery in this growing population of patients not experiencing optimal postoperative results.

NIH Research Projects · FY 2026 · 2023-09

Abstract The aged brain is thought to be more vulnerable to stresses than its young counterpart, and different in its coping with neuroinflammation and ability to repair an injury. A better understanding of the brain aging process will provide valuable information. This knowledge enables one to mitigate age-related declines in cognitive, emotional, sensory, and motor functions. Such information may also promote effective strategies for treating age-related neurodegenerative diseases, such as Alzheimer’s disease (AD). The brain is composed of multiple types of non-neuronal cells besides neurons, and each type seems to undergo unique age-related changes following its genetic program. Oligodendrocytes (OLs), a major glial cell population, form myelin sheaths, essential for rapid axonal conduction in the central nervous system (CNS). OLs also provide metabolic and nutritional support to neurons and contribute to other homeostatic regulations for axonal communication. Recently, our OL-specific transcriptomic analyses revealed that IL-33, a member of the IL-1 family known to contribute to neural circuit refining and neural repair, is increasingly expressed in OLs with age. Consequently, at one year of age, OLs become the predominant source of IL-33 (> 90% of all IL33-expressing cells) in the mouse CNS. Interestingly, IL-33 genetic variations are correlated with the risk of AD in patients, and higher levels of IL-33 in the brain significantly benefited amyloid plaque clearance in mice. Given the critical functions of IL-33, it is crucial to identify detailed source cell-specific mechanisms of IL-33 in the aged brain. To understand how OL-derived IL-33 shapes brain aging and AD-like disease progression, we will employ mouse genetic tools that allow OL-specific IL-33 conditional knockout (cKO) or overexpression. We will examine the effects of those genetic manipulations on OL survival and myelin maintenance in the aged brain. Moreover, these IL33-related genetic manipulations will be applied to a mouse model of AD (APP/PS1), and we will determine whether OL-derived IL-33 regulates AD-like diseases and cognitive deficits, as well as microglia-mediated clearance of beta-amyloid (Aβ) deposits. The same genetic manipulations will also be used on astrocytes; thus, the relative importance of OL-derived IL33 will be compared with astroglial IL33. If successfully conducted, this study will advance our understanding of cell-cell interactions, especially those mediated by IL-33 in brain aging and during AD progression. Our results may promote the development of a therapeutic strategy with an oligodendroglia-targeted approach and identify related molecular mechanisms and targets for treating AD patients.

NIH Research Projects · FY 2025 · 2023-09

Abstract: Periodontitis is the second most prevalent but preventable dental disease affecting over 64 million Americans and responsible for tooth loss, functionality limitations, pain, and poor quality of life. Thus, early diagnosis and preventive therapeutics are imperative in clinical practice to prevent disease initiation and progression. However, by the time dentists can observe the first bone loss patterns in radiographs to diagnose periodontitis, 30-50% deterioration (periodontal bone damage) has already occurred, which is not visible to human eyes. Clinical decision support systems are designed to identify high-risk periodontitis patients for prevention; however, they are not widely used in clinical practice because of the suboptimal prediction performance and lack of diverse predictive features (early bone loss lesions) for prediction. Therefore, there is an unmet need for a tool that can detect early bone loss patterns invisible to human eyes to alert dentists for early diagnosis and preventive care. Dr. Patel has developed an artificial intelligence (AI) empowered prediction model for periodontitis that utilizes more than 150 distinct variables (e.g., social determinants of health, medical records, lab reports, CDC census data, financial data, etc.) for prediction, which aren't well understood in the existing literature. However, this model lacks dental imaging data such as bone pattern, bone density, pixel intensity, and other imaging predictive features, which have a high potential to improve prediction accuracy. The early bone mineral changes in alveolar bone for early diagnosis have been studied in biological studies; however, the transition of these findings at the chairside is limited. AI and computer vision can bridge this gap and help identify early bone loss patterns from radiographs invisible to human eyes. Therefore, the objective of this project is to develop three automated computer vision algorithms: 1) to improve the extraction of diagnostically meaningful information from periapical radiographs, 2) to determine the extent of bone loss information from radiographs, and 3) build a prediction model to identify early bone loss patterns from radiographs before disease initiation and progression. Enhanced and consistent radiographs will improve diagnostic accuracy & reduce radiographic exposure, automatic bone loss measurement will reduce diagnostic discrepancies, and early bone loss detection will identify high-risk patients to take preventive approaches. The candidate, Dr. Patel's goal is to become an independent PI in dental informatics and develop cutting-edge technologies to generate practice- based evidence (using data-driven methods) to improve patient care and outcomes. A funded K08 proposal will allow Dr. Patel to develop the skills necessary to complete the proposed research (training in computer vision & radiology) and become an independent research scientist (training in didactic mentoring, lecturing, & grantsmanship). Dr. Patel has formed a team of five mentors with expertise in clinical dentistry, computer vision, radiology, and periodontology to provide high-quality, diverse scientific, collegial support and state-of-the-art facilities to ensure the successful completion of this proposed career development goals and research program.

NIH Research Projects · FY 2024 · 2023-09

Abstract: Mitochondrial function underlies a broad spectrum of health and disease conditions that range from metabolic rate regulation, thermogenesis, to inborn genetic disorders, cancer, aging, cardiac and muscle dysfunction, diabetes and obesity. Using oxidative phosphorylation, the mitochondria pump protons from the matrix to the intermembrane space to generate a proton gradient. The protons return to the matrix via the ATPase complex to generate ATP that supports the life of the organism. About 20-50% of protons leak back to matrix but do not drive ATP production and this is called proton leak or uncoupling. The traditional proton leak measurement by Clark type oxygen probe and Seahorse assay use oxygen consumption rate (OCR) as a readout. These OCR based proton leak measurements cannot exclude “proton slip”, which is defined as respiratory chain activity that consumes O2 and transfers electrons without extruding protons out of the membrane. There is a knowledge gap of how to assess the specific mitochondrial proton leak and slip. To address this gap, I established a direct method for measuring the accurate proton leak and eliminating proton slip by exposing mitochondria to a pH gradient stress in saponin permeabilized cells with mt-cpYFP, a mitochondrial targeted pH sensitive indicator. The buffers lack metabolic substrates, thus making the cells energetically inactive, allowing separation of the physical property of the mitochondrial inner membrane from energetically active processes such as ion pumping. Using this approach, I have revealed that the adenine nucleotide transporter 1 (ANT1) has an under-appreciated role in supporting an excessive proton leak. With this novel mitochondrial proton leak measuring method, in this R35 MIRA proposal, we propose to 1) Compare mitochondrial proton leak between different cell types from the mt-cpYFP transgenic mouse; 2) Dissect the molecular basis of ANT1-mediated mitochondrial proton leak; 3) Screen for new drugs/compounds to prevent or augment the mitochondrial proton leak. Completion of this study will provide key mechanisms of mitochondrial proton leak and will provide a novel group of drugs that modulate mitochondrial function involved in the metabolic homeostasis of these diverse health processes.

NIH Research Projects · FY 2024 · 2023-08

ABSTRACT Understanding the mechanisms underlying GPCR signaling is crucial in order to fully comprehend their role in physiology and pathophysiology. In addition to canonical second messengers (cAMP, cGMP and IP3) and β- arrestin signaling, small GTPase proteins, such as Rho GTPases are largely involved in GPCR-mediated signal transduction. Guanine nucleotide exchange factors (GEFs) convert Rho GTPases from an inactive (GDP-bound) state to an active state (GTP-bound). Rho-GEFs can be activated by Gq, G12/13 and Gs proteins. However, currently there is no evidence that Gi/0-WT can directly activate RhoGEFs. PDZ domains are structural protein domains that recognize simple linear amino acid motifs often at the protein C-terminal (C-motif). RhoA, activated by PDZ-RhoGEFs, has important signaling roles, by activating phospholipase D (PLD) and transcription factors. Cannabinoid receptor CB1, an abundantly expressed GPCR that mainly couples to Gi/o, has a EAL C-motif (last three amino acids) that binds PDZ class III proteins, including PDZ-RhoGEF. Our central hypothesis is that CB1 receptor activation, in addition to engaging cAMP inhibition and β-arrestin pathways, initiates an additional signaling mechanism downstream to PDZ-RhoGEF leading to activation of RhoA and subsequent activation of PLD. Activation of PLD generates two distinct second messengers, phosphatidic acid, which activates the mTOR pathway, and choline, which activates Sigma1 receptors. We will use a multidisciplinary approach, combining state-of-the-art molecular and pharmacological approaches for a comprehensive investigation of the signaling pathways elicited by activation of C-motifs of CB1 receptor and identification of pathways-selective ligands. We will use receptor and PDZ-RhoGEF mutations, measurements of second messengers levels (cAMP, choline and phosphatidic acid) as well as live imaging of PLD activation. We provide solid preliminary results supporting the feasibility of the project and the ability of our team to complete the work proposed. The project has two aims: Aim 1. Investigate the role of PDZ-binding domain in CB1-induced signaling; experiments are designed to characterize intracellular cascades activated by CB1-motifs (CB1-PDZ binding domain. Aim 2. Investigate the role of PDZ-RhoGEF/RhoA pathway in CB1 receptor signaling in cultured primary neurons and in vivo. The successful completion of this project will increase the current knowledge of GPCR signaling and will serve as a basis for further development of ligands selectively targeting this pathway.

NIH Research Projects · FY 2024 · 2023-08

ABSTRACT Surveillance of many human pathogens relies primarily on clinical diagnosis of individuals who seek testing or care. In most contexts the number of unobserved infections is unknown, which poses a challenge in effectively mitigating transmission. Infections may go unobserved for a number of reasons, including mild and asymptomatic infections not captured by clinical surveillance as well as symptomatic infections not captured due to factors such as barriers in access to healthcare or limited testing resources. This is true for Vibrio cholerae, a pathogen responsible for an estimated 3 million cholera cases and 100,000 deaths each year. Previous studies in Bangladesh have shown that unobserved infections account for at least half of V. cholerae infections and that individuals with asymptomatic infection shed less V. cholerae in their stool compared to symptomatic cholera. The extent to which this impacts our ability to prevent and control outbreaks, and whether these findings apply to other populations, is unknown. In this study we aim to address this gap in our knowledge by investigating the contribution of unobserved infections to cholera burden and transmission in multiple populations. We propose to do this by estimating the magnitude of V. cholerae infections that go unobserved by comparing patterns in clinical and serological data across various endemic and epidemic locations (aim 1), examining the degree to which individuals with mild symptoms who commonly go unobserved shed V. cholerae bacteria that may be transmitted (aim 2), and estimating the risk of infection following exposure to asymptomatic household contacts (aim 3). We expect to find high levels of unobserved infections, particularly in areas with barriers in access to healthcare. We also anticipate that mild/asymptomatic infections will be associated with bacterial shedding and ongoing transmission, but to a lesser extent than cases with more severe symptoms. Alternatively, since people with mild/asymptomatic infections are less likely to receive antibiotics and often continue to go about their daily routines, we may find increased bacterial shedding and secondary infections in these cases. In either scenario, these results will provide important insights into the degree to which unobserved infections contribute to V. cholerae persistence and spread, as well as their role in our efforts to mitigate outbreaks. Our long-term goal is to better understand the underlying host, microbial, and socio-demographic factors that lead to infections going unobserved so that we can design more effective prevention and control strategies.

NIH Research Projects · FY 2024 · 2023-08

ABSTRACT In Alzheimer’s Disease (AD), and to a lesser extent in non-demented individuals, deposition of amyloid β (Aβ) is found around cerebral vessels, a condition called Cerebral Amyloid Angiopathy (CAA). It is likely that the development of CAA, and consequently neurovascular unit (NVU) dysfunction, in AD may result from failure of Aβ clearance pathways. Importantly, all Aβ clearance pathways involve movement of Aβ towards the vasculature for elimination. Cerebrovascular endothelial cells (EC), which line all brain vessels, are the gatekeepers of the brain and are responsible for the maintenance of cerebral homeostasis through the blood-brain barrier (BBB). The development of CAA severely impacts brain health since it results in EC death, BBB breakdown, microhemorrhages, parenchymal Aβ accumulation, and is one of the earliest triggers for AD progression. EC dysfunction also affects other cells of the NVU, such as astrocytes or microglia, through finely regulated communication mechanism between these cells. Therefore, CAA-mediated EC dysfunction may also precipitate neuroinflammation in AD. Our preliminary data suggests that EC mitochondria serve as important sensors of Aβ damage. Previous studies have shown that mitochondria are responsible for the activation of inflammatory pathways through the release of mitochondrial danger associated molecular patterns (mtDAMPs), including mitochondrial reactive oxygen species (mtROS), due to formation of the pathological mitochondrial permeability transition pore (mPTP). Both cerebrovascular dysfunction and inflammation are shown early in disease pathogenesis, suggesting that early EC (and thus, BBB) dysfunction may drive and perpetuate AD pathology also by triggering widespread neuroinflammation. Here, we will test the hypothesis that Aβ induces mitochondrial dysfunction in cerebral ECs through deleterious alterations in mitochondria function (mtROS and mPTP opening) and that these alterations further contribute to neurovascular dysfunction and inflammation in CAA and AD. This proposal seeks to understand the role of alterations in cerebral EC mitochondrial health and bioenergetics, specifically focusing on the modulation of mtROS and mPTP, on human EC inflammatory activation due to Aβ (Aim 1; K99: Y1 and Y2). Moreover, we want to assess how Aβ-induced cerebral EC inflammatory mediators contribute to glial mitochondrial dysfunction and activation, through the modulation of mtDAMPs, using a human iPSC-derived glial cell cultures (Aim 2; K99: Y2 and R00: Y1). Finally, using an animal model of amyloidosis presenting CAA and gliosis (TgSwDI), we will test the hypothesis that Aβ-induced endothelial mtROS and mPTP mediate neuroinflammation and cognitive decline, by treating the animals with a mtROS scavenger or depleting their ECs of CypD, the main mPTP regulator (Aim 3; R00: Y1-Y3). The proposed experiments will provide a novel understanding of the role of vascular mitochondria as initiators of inflammation in AD and CAA, potentially resulting in the development of new disease modifying therapeutic strategies.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY/ABSTRACT Adolescence is a key risk period for depression and anxiety, and adolescent-onset psychopathology is predictive of poorer health and life outcomes. Functional connectivity (FC) networks, which reflect trait-like brain functioning, have emerged as a promising biomarker to inform diagnosis and intervention of psychopathology. However, despite findings of FC differences between clinical and control groups, particularly in resting state (RS) networks, there has been minimal clinical translation. A key limitation is that qualitative network heterogeneity between- and within-individuals threatens the ability to draw inferences valid at the individual level. At the between-person level, qualitatively distinct networks across individuals limit the ability of group-averaged networks to validly reflect each individual. If group-level networks do not reflect individuals, behavioral inferences drawn from them will not apply to the individual. Our preliminary work and previous precision imaging studies provide evidence of this limitation by demonstrating FC network heterogeneity across individuals. However, the generalizability of group networks to individuals is yet to be tested empirically. This proposal will assess group-to-individual generalizability of adolescent RS networks and examine the ability of data-driven subgroups of similar individuals to address the limitation of heterogeneity (Aim 1). At the within-person level, FC variability across a single scan also threatens the validity of FC networks. If FC means and covariances vary with time across a scan (i.e., are not stationary), a static (time-invariant) network would not validly reflect network processes across the scan. This proposal will estimate dynamic FC to assess stationarity of adolescent RS networks to determine the validity of static networks (Aim 2). For both aims, this proposal will use a large 11–12-year-old sample from the Adolescent Brain Cognitive Development study. The proposal will determine the levels of data aggregation (group, subgroup, or individual) and time precision (static or dynamic) necessary for individual-level inferences from FC networks. Findings will be critical for the ultimate goal of using FC networks for clinical translation, which requires individual-level prediction. We will then use RS network features that are precise to individuals to predict depression and anxiety outcomes in adolescents (Aim 3), building a foundation with increased potential for clinical translation. The training plan to achieve the proposed project was developed in collaboration with a team of relevant experts that consists of formal coursework, workshops, and applied research activities. Specifically, I will develop expertise in fMRI research and analysis methods, machine learning approaches for subgroup identification, and idiographic (person- centered) methods necessary to complete the research proposal. Training will emphasize development toward my goal of becoming an independent investigator in clinical neuroscience who studies individually precise associations between neural functioning and adolescent psychopathology.

NIH Research Projects · FY 2025 · 2023-08

Abstract We propose to study whether targeting both the transcription factor ZNF362 and cytoskeletal non-muscle myosin IIA and IIB (NMIIA&B) will promote better axon regeneration and functional recovery after spinal cord injury (SCI) than either approach alone. After SCI, severed axons fail to regenerate largely because of the reduced intrinsic growth capacity of adult CNS neurons and the poor environment for axon extension. Treatments to recover paralysis and other lost functions are not available and patients with SCI are often permanently disabled. Many genes have been determined to control the regrowth failure of mature neurons, but none have been translated to clinical use. There is a persistent need to identify better gene targets and therapeutic strategies. ZNF362 is highly expressed in the CNS, but its major function in mammals remains largely unknown. The PI’s group generated ZNF362 conditional knockout (cKO) mice and designed novel small peptides to block ZNF362 function selectively. Our pilot studies suggest that ZNF362 strongly suppresses the growth capacity of CNS neurons in adult rodents. After CNS injury, various inhibitory molecules around the lesion activate neuronal RhoA, which, in turn, activates cytoskeletal NMIIA&B to condense actin filaments and restricts microtubule protrusion and axon elongation. Because neuronal cytoskeleton is the major machinery to drive axon growth and the converging targets of multiple signaling pathways that control axon growth, manipulating neuronal cytoskeleton is also very attractive to promote significant CNS axon regeneration. We hypothesize that inhibiting both ZNF362 and NMIIA&B represents a dual approach for enhancing neuronal growth capacity and reducing environmental inhibition around the lesion. We propose to dissect potential critical roles of ZNF362 and NMIIA&B for controlling regrowth of mature CNS neurons and to develop novel and effective strategies for promoting CNS axon regeneration. We aim to stimulate robust axon regrowth and functional recovery in SCI rodents by suppressing these genes using cKO mice and new selective antagonist peptides designed in our lab. In Aim 1, we will study whether transgenically deleting ZNF362 and NMIIA&B acts synergistically to promote axon regeneration and recovery in adult mice with SCI. Aim 2 proposes to determine whether blocking individual ZNF362 and NMIIA&B signals pharmacologically with novel selective antagonists promotes axon regeneration and recovery in adult rodents with SCI. In Aim 3, we plan to develop combination therapies that block both ZNF362 and NMIIA&B signals, aiming to yield robust axon regrowth and functional recovery in adult rodents with SCI. Based on the promising results of our pilot studies, we anticipate that our novel regenerative strategies will significantly advance our ability to treat SCI. If our peptides are successful with rodent SCI models, we plan to move this work to peptide safety assessments and further translational studies (e.g., cervical/chronic SCI and clinical trials). Therefore, this project may facilitate identifying novel molecular targets for CNS repair and highly effective strategies for treating CNS lesions.

NIH Research Projects · FY 2025 · 2023-08

ABSTRACT There are ~40 million people world-wide infected by the Human Immunodeficiency Virus Type 1 (HIV-1, commonly referred to as HIV). As currently there is no cure, antiretroviral treatment is the primary treatment option. Yet antiretroviral treatment eventually fails over time due to the development of drug resistance. We will develop new computational tools for forecasting HIV evolutionary trajectories under therapeutic selection pressure leading to drug resistance, using high resolution all atom molecular dynamics simulations together with physics-based machine learning models of sequence co-variation. The computational studies will be complemented by structural, biophysical, and virological studies on two HIV protein multimeric targets: HIV integrase (IN), and capsid (CA). The modeling and experiments will be employed in an iterative manner, with the experimental results being used to validate, parameterize and improve the models; and the molecular dynamics simulations used to guide new experiments and also to develop new tools for high resolution cryo-EM refinement of multiple binding modes of HIV inhibitors and interfacial solvent. The common theme of our proposed work is to provide structural interpretations for the observed fitness and resistance effects of mutations, with the goal of developing holistic structure-function models which can be used to predict viral mutation trajectories under drug selection pressure and give a mechanistic explanation for them. There are three specific aims: (1) determine the physical mechanisms underlying mutational epistasis under varied drug environments, and use MD simulations and virological data to parameterize drug specific landscapes for HIV IN and CA under a novel theoretical framework; (2) use high resolution, large scale alchemical molecular dynamics free energy simulations based on advanced sampling methods to analyze the effects of protein mutations on the stability of protein-protein interfaces that constitute the intasome and the capsid particle assemblies; (3) determine the molecular basis for multiple binding modes of inhibitors of HIV IN, and the role of solvation in the strong binding of these inhibitors. These aims seek to achieve a molecular understanding of the cooperative effects (epistasis) of multi-residue mutation patterns on the binding of inhibitors to their viral protein targets (IN, CA) and their effects on the stability of the multimers (intasome and capsid particle). We anticipate that this work will lead to the development of surveillance tools to forecast the response of viral systems to the selection pressure of antiviral therapeutics.