University Of Nebraska Lincoln

NIH Research Projects · FY 2026 · 2023-12

Project Summary/Abstract The broad, long-term goal of the proposed work is to provide children with severe speech and physical impair- ments (SSPI) access to effective augmentative and alternative communication (AAC). AAC devices provide a means of communication for those who find speech communication inefficient or ineffective due to disability. For children with SSPI, the lack of AAC access options has a devastating impact on quality of life, well-being, and medical care. By translating brain activity into communication device control, brain-computer interfaces (BCIs) for AAC (BCI-AAC) can support access to communication for those with SSPI who find existing methods of AAC inaccessible, fatiguing, or difficult to learn, and expand AAC options to facilitate multimodal device control. P300- based BCI-AAC devices use brain activity associated with attention to a target item and provide an encouraging avenue for children's AAC access due to the simple control task and short training times. However, P300-BCI- AAC research primarily focuses on developing spelling-based interfaces for adults, with limited research on chil- dren largely focusing on adolescents (aged ≥13 years), even though cortical maturation impacts the P300 brain signal. Recent works that employ implementation science frameworks and AAC experts have identified a para- mount need to build on prior foundational research and extend P300-BCI-AAC access to those in middle child- hood who may be unable to spell. Problematically, the development of picture-based P300-BCI-AAC systems for children aged 8–12 years remains largely unexplored. Thus, this project will break new ground in (a) picture- based P300-BCI-AAC development and (b) clinical translation, by determining initial levels of picture-P300-BCI- AAC performance for both healthy children and those with SSPI in middle childhood. Further, it will establish factors impacting BCI-AAC accuracy and children's design preferences. Project outcomes will be vital to achieve accelerated BCI-AAC success for individuals and the NIDCD's plan for patient-oriented implementation. Aim 1 will determine initial picture-based P300-BCI-AAC performance across two sessions, which will promote comfort, as BCI-AAC equipment may seem unfamiliar. Aim 2 will provide new knowledge of brain activity underlying P300-BCI-AAC access and how a range of patient-oriented factors impact accuracy to inform the development of assessment tools that can accelerate training times. Finally, to promote engagement, Aim 3 will evaluate and establish the P300-BCI-AAC design preferences of children via the use of a novel BCI-AAC design creation application. Project findings will have potential to improve communication abilities of children with SSPI via BCI- AAC and help decrease the impact of disability and risk of preventable adverse medical events (e.g., incorrect drug administration) while promoting quality of life and social participation. Outcomes will inform larger-scale investigations to develop assessment materials and inform future research to elucidate how children with SSPI learn BCI-AAC control in natural environments, alongside how display designs impact performance.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY/ABSTRACT Alcohol use and destructive couple conflict, including intimate partner violence (IPV), represent dual public health threats for sexual and gender minorities (SGM), who experience higher rates of these problems than their cisgender, heterosexual counterparts. Although a broader literature links alcohol use to increased rates of destructive couple conflict, very little work has examined these associations in SGM couples. Here, we draw on alcohol myopia theory and propose that increased daily alcohol use will be associated with higher same-day levels of destructive conflict among SGM couples. Further, we predict that greater drinking to cope motives and higher levels of internalized minority stress (i.e., fear of rejection, worry about concealment, internalized homophobia/transphobia) assessed at baseline will exacerbate the effects of daily minority stress (exposure to discrimination, harassment, and stigmatization) on alcohol use and subsequent destructive conflict. Importantly, these processes will be examined in a sample of SGM couples residing in the rural Midwest—a population that is critical to study because of the increased types and frequency of stigma encountered by SGM individuals living in rural areas as compared to more urban locations. Finally, we test the hypothesis that the detrimental impact of minority stress on alcohol use and destructive couple conflict will be mitigated by greater (a) social support from interpersonal relationships, (b) psychological sense of connection with the LGBTQ+ community, and (c) psychological sense of community with one's rural Midwestern neighborhood. Participants will be 200 SGM couples (i.e., both individuals identify as lesbian, gay, bisexual, transgender, and/or queer, and are in a committed intimate relationship). Couples will be recruited from the Midwestern LGBTQ+ Research Registry, established by the MPIs. Our proposed models will be tested using intensive daily diary methods employed over 60 days to track daily experiences of minority stress, alcohol use, and destructive conflict. Findings from this project will provide novel data about the conditions under which daily minority stress contribute to elevated levels of destructive couple conflict via increased alcohol use by rural SGM couples. Our examination of support and community-based resiliency factors will highlight potential points of intervention that can be targeted to interrupt the harmful effects of minority stress on alcohol use and subsequent intimate partner conflict and violence.

NIH Research Projects · FY 2024 · 2023-09

Project Summary/Abstract Earlier onset of a psychiatric disorder is associated with an increased likelihood of meeting criteria for additional disorders and impaired functioning across the lifespan. Thus, efforts aimed at preventing or reducing the development and maintenance of psychopathology early in life are of great value, especially when focused on identifying early modifiable risk factors and mechanisms most salient to its development. The overall objective of the present study is to investigate how the parent-child relationship across infancy and toddlerhood – a sensitive period consisting of increased neuroplasticity and sensitivity to the quality of the caregiving relationship – ultimately impacts the development of transdiagnostic psychopathology and its underlying neurocognitive mechanisms. Additionally, to address the dearth of research on the importance of fathers on children’s development, we will examine the unique contributions of both mothers and fathers in the development of children’s psychopathology and its underlying neurocognitive mechanisms. Based on prior theory and empirical evidence, we hypothesize that preschool executive control will be a mechanism through which the quality of both the mother-child and father-child relationship in infancy and toddlerhood impacts the development of the p-factor, a general shared dimension of symptoms that underlies nearly all forms of psychopathology. To test this hypothesis, the integrative, comprehensive construct of mutually responsive orientation (MRO) will be observed in both the mother-child and father-child relationships during infancy and toddlerhood as predictors of child executive control in preschool and the general factor of psychopathology at school age. MRO captures a dyadic level of relationship quality comprised of coordinated routines, harmonious communication, mutual cooperation, and emotional ambience (high MRO) and maladaptive qualities of the relationship such as hostile communication, lack of responsiveness, and high levels of negative affect (low MRO). Study aims will be pursued in an established sample of 159 families (mother, father, and child) who have completed observational paradigms when the target child was 1 and 2 years of age. Preschool executive control will be measured using a comprehensive, laboratory battery of nine executive functioning tasks at age 5 years. Child general psychopathology will be measured using both mother and father reports of well-validated, reliable, and developmentally appropriate psychopathology questionnaires of child internalizing problems, externalizing problems, and emotion dysregulation when participating children turn 7 years of age. Findings are expected to identify modifiable targets for developmentally informed prevention and intervention efforts aimed at reducing executive dysfunction and the subsequent development of nearly all forms of psychopathology across the lifespan.

NIH Research Projects · FY 2025 · 2023-07

Project Summary Both naturally occurring and laboratory protein modifications are important fields of research since proteins are central to virtually every biological process. On one hand, protein post-translational modifications (PTMs) are essential for proper functioning of an organism and are associated with many diseases. It is of great significance to study PTMs in order to develop new therapeutic agents. On the other hand, continuing advances in noncanonical amino acid (ncAA) mutagenesis has provided powerful tools for the manipulation and study of protein function. This MIRA application seeks to merge investigations on both fronts of these research areas. The first general area of this MIRA project focuses on the co-translational modification of protein through genetic code engineering. The long-term goal is to explore innovative strategies for ncAA mutagenesis through reprogramming the genetic code. Our immediate and unique focus is on reprogramming codon language with quadruplet codons (Qcodons), although triplet nonsense codons are the predominant ones used in the field of genetic code expansion. Built on our pioneering efforts on tRNA engineering, we propose a new direction to improve Qcodon decoding efficiency through the identification and implementation of recoding signals. The use of recoding signals can also significantly mitigate a major concern over undesirable readthrough of endogenous stop codons with nonsense suppression-based noncanonical amino acid (ncAA) mutagenesis in live cell studies. We will also apply a Qcodon-dependent and ncAA-mediated control strategy to the development of HIV-1 vaccines, which represents a novel direction that was first demonstrated by my group. Such strategy will also be applied to the generation of vaccines against other pathogenic viruses or bacteria in the future. The second general area of this MIRA project is to investigate the role of protein tyrosine O-sulfation (PTS) in mammalian cell biology. Our initial efforts will focus on PTS of chemokine receptors as the first step of our long-term efforts to study the effects of PTS on receptor signaling in general. Furthermore, we seek to develop therapeutic agents targeting sulfated receptors. These two innovative projects have their own goals and significance, but are unified under our expertise in chemical/synthetic biology and are partially associated with each other. The long-term goal of my laboratory is not only to develop innovative and meaningful chemical/synthetic biology tools, but also to employ these tools to gain insights into biomedical processes for the development of novel therapeutics.

NIH Research Projects · FY 2026 · 2023-07

PROJECT ABSTRACT Nicotine is the primary addictive constituent in tobacco. For an individual, chronic tobacco use is associated with a significant increase in heart disease and many forms of cancer. Increasingly, researchers are documenting health issues associated with vaping flavored e-juice containing nicotine. There are some important sex differences in chronic nicotine use and its health consequences. Women on average are impacted more by environmental and sociocultural factors, take less time to become nicotine dependent, make fewer attempts to quit, abstain for less time, relapse at higher rates, and benefit less from nicotine replacement therapy. Enormous gains to the individual and society would come from a more complete understanding of factors that contribute to nicotine use and misuse. Through Pavlovian conditioning processes, environmental stimuli associated with drugs such as nicotine can become powerful modulators of drug-seeking behavior. Another related and important factor in the development of chronic nicotine use is the pharmacological effects of nicotine serving as an internal (interoceptive) stimulus that enters into a conditioned association with other reinforcers (e.g., peer acceptance, alcohol, work breaks, stress relief, etc.). Thus, smokers/vapers over time develop a rich interoceptive conditioning history with the stimulus effects of nicotine. Our long-term objective is to understand how this conditioning history alters the trajectory of use and misuse in individuals and, in doing so, provide new clues on how to improve cessation programs. To this end, we recently developed an innovative new approach using rats that merges interoceptive conditioning of intravenous (IV) nicotine with its later self-administration. When IV nicotine is repeatedly paired with an appetitive reinforcer (i.e., sucrose), this conditioning history dose- and sex- dependently increases nicotine self-administration (21 to 126%)—an effect more profound in female rats. Leveraging the methodological strengths of this new approach, we address three Aims in the present proposal. Aim 1 will assess whether the need to recall the learning after a retention interval alters the strength of the conditioned reinforcing effects of nicotine. Aim 2 will determine whether the addition of a weak reinforcer during self-administration interacts with nicotine and its newly acquired conditioned reinforcing effects. Aim 3 will examine whether an external (exteroceptive) stimulus will compete with the interoceptive nicotine stimulus for conditioned reinforcing value, thus weakening its later impact on nicotine intake. A fundamental understanding of such factors that influence the effect of interoceptive conditioning with nicotine will refine and advance our theoretical models. These advancements will provide foundational research for other basic scientists and clues for clinical scientists and healthcare providers working to improve approaches to treating nicotine use disorder.

NIH Research Projects · FY 2026 · 2022-12

Viral manipulation of mitotic and antiviral signal transduction determines the outcome of infection, but remains poorly understood. To address this knowledge gap, our laboratory studies a family of protein kinases comprised of homologs widely expressed in poxviruses and in all multicellular eukaryotes. The long term goal of our research is to determine how poxviruses usurp and redirect signaling cascades governing mitotic and host defense effectors responsive to foreign DNA. Mammalian poxviruses express two proteins, B1 and B12, which are homologous to each other and to three eukaryotic protein kinases named vaccinia related kinases (VRKs). Comparative studies of B1 and VRK1 revealed that both enzymes directly modify the cellular protein BAF. Importantly, BAF acts as both a mitotic regulator and antiviral effector by binding and compacting dsDNA, a property that is inactivated via phosphorylation by B1 or VRK1. Our new data argue that B1 and cellular VRKs co-regulate other pathways as well, including an antiviral pathway activated by the B12 protein. Our data indicate that B12 directs strong repression of vaccinia DNA replication via partly unknown mechanisms also governed by B1. Intriguingly, B12 is a nuclear poxviral protein and a non-catalytic kinase or `pseudokinase', which are of key innovative importance for this proposal. Pseudokinases are members of the pseudoenzyme family, about which little is known in viruses. It is our central hypothesis that vaccinia B1 and B12 form a novel signaling axis that supplants and redirects cellular VRK pathways regulating BAF and other VRK substrates such as histones and the HUSH (Human Silencing Hub) complex. To test our hypothesis, we propose three aims. AIM 1) Determine how B1 and B12 remodel VRK1-responsive signaling during poxvirus infection. This Aim tests the hypothesis that B12 interacts with VRKs in the nucleus, thereby altering H2A, BAF, and HUSH regulation. Characterization of B12 interaction with BAF, B1, and cellular VRKs in vitro and in cultured cells will be achieved. AIM 2) Determine the molecular mechanisms governing B12 repression of poxvirus DNA replication. This Aim tests the hypothesis that the B12/VRK1 complex plays key roles in the mechanism of B12 signaling dysregulation. Structure/function analysis of B12 through targeted mutational analysis, novel loss of function screens, and investigation of B12 phosphoregulation are outlined in this Aim. AIM 3) Determine how viral/cellular pseudokinases mediate repression of the poxvirus lifecycle and converge with protein phosphatase signaling. This Aim will test the hypotheses that B1 and VRK2 kinases regulate B12 via direct phosphorylation while VRK3 and the phosphatase PP2A control dynamic regulation of B1/VRK1 substrates, leading to manipulation of downstream antiviral responses. The completion of this work will: fill gaps in our understanding of poxvirus manipulation of nuclear processes, yield broadly relevant insights to the field of kinase-pseudokinase biology, and provide needed information of how mitotic and antiviral signaling interweave.

NIH Research Projects · FY 2024 · 2022-09

Project Summary The commercial sexual exploitation of children (CSEC) is a public health crisis in the U.S. To date, however, we know little about how to prevent CSEC. The Set Me Free Project has developed the READY to Stand (RTS) curriculum consisting of four, 45-minute modules implemented to high school students and school personnel. The dynamic RTS provides students with psychoeducation on CSEC, healthy relationship skills training, identification of safe people and resources, and programming components to enhance valuing of self and others. As part of the proposed project, the RTS will be enhanced with two additional 45-minute modules that focus on bystander intervention training in situations of CSEC and shifting school and community norms to be intolerant of all forms of violence, including CSEC. Despite its potential for reducing CSEC, the RTS has never been evaluated. The overarching goal of this multi-stakeholder collaboration, including researchers, educators, practitioners, and youth, is to conduct both a process and rigorous outcome evaluation of the RTS with an eye toward widespread dissemination to other communities in the U.S., if deemed effective. The implementation site includes five traditional three alternative high schools in Des Moines Public Schools (DMPS); students are largely racial/ethnic minority (65%) and low-income (76%). Component A (Years 1-2), the refinement and planning phase, includes the creation of a Research Advisory Board (RAB) comprised of researchers, practitioners, educators, caregivers, and youth; development of the two supplemental 45-minute modules on bystander intervention and social norms for the RTS (student version) and enhancement of the 90- minute RTS for school personnel version; creation of psychometrically validated survey instruments to measure CSEC perpetration and victimization and related CSEC constructs; and the conduction of an open pilot trial of the RTS with one traditional high school and one alternative high school in DMPS (n=878 students; n=78 school personnel). Component B (Years 3-5) comprises the rigorous evaluation phase and includes a quasi-experimental study in which the remaining four traditional high schools and two alternative schools are randomly assigned to the treatment or wait-list control conditions. Students (n=7,241) will complete baseline and 6-, 12-, and 18-month follow-up surveys to test the hypothesis that participation in the RTS will lead to reductions in CSEC perpetration (primary outcome) as well as reductions in CSEC victimization and teen dating violence and sexual assault victimization and perpetration and increases in bystander intervention in CSEC situations (secondary outcomes). We will assess mediators (e.g., increases in CSEC knowledge, CSEC bystander efficacy) and demographic moderators of program impact and conduct an in-depth process evaluation that includes data from both students (n=7,241) and school personnel (n=396). This study represents the first ever rigorous evaluation of a CSEC prevention program. Findings from both Components A and B will be widely disseminated to diverse audiences with RAB engagement.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY During young adulthood, an estimated one in five women experience sexual assault. We seek to reduce this violence by motivating young adults to intervene with their peers to prevent sexual assault—an approach known as bystander intervention. Current bystander training is conducted in group sessions involving education about how to recognize and intervene in response to sexual risk situations. Although successful in changing knowledge and attitudes about sexual assault prevention, evaluations of these programs have rarely focused on changing actual bystander behaviors. Further, while bystander alcohol use is common in sexual risk situations, and undermines intervention attempts, alcohol consumption by bystanders is not explicitly targeted in existing intervention training programs. To address these gaps, we will conduct a RCT comparing the efficacy of: 1) our recently developed bystander intervention, Motivate-the-Bystander (MTB), 2) MTB with an MI alcohol component (MTB+ALC), and 3) an attention control condition for reducing alcohol use and increasing bystander behaviors in response to sexual risk. Bystander behaviors will be assessed observationally during a virtual reality-based house party at 2 months post intervention. Participants’ bystander behaviors, alcohol use, and relevant contextual variables will be assessed with a measurement burst design using electronic daily diaries at baseline and 3, 6, and 9 months post intervention. We expect that, compared to MTB alone and the control condition, MTB+ALC will produce significantly greater reductions in overall drinking and increases in prosocial bystander behaviors in a diverse sample of 450 young adults who are heavy drinkers. If our hypotheses are confirmed, results will support the use of our combined MI-based bystander-alcohol intervention as an effective means of reducing drinking and motivating bystander behaviors among those at highest risk for sexual violence.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY High-altitude animals have evolved the ability to survive and function under conditions of oxygen deprivation (hypoxia) that mimic disease states in humans. Identifying evolved mechanisms of hypoxia adaptation in mammals that are long-term high-altitude natives can therefore yield discoveries of biomedical relevance while also providing general insights into the evolution of complex traits. A number of wild rodent species inhabit far more extreme altitudes than Tibetan and Andean humans and also represent far more tractable subjects for experimental approaches that involve genetic crosses and invasive physiological manipulations. This project integrates genomics and experimental physiology to dissect the mechanistic basis of adaptive enhancements of whole-animal performance in hypoxia in extreme high-altitude rodents. The experiments compare high- and low- altitude populations of two species: the deer mouse (Peromyscus maniculatus), which has the broadest altitudinal range of any North American mammal (sea level to 4350 m), and the Andean leaf-eared mouse (Phyllotis vaccarum), an extremophile species that holds the record as the world’s highest dwelling mammal and that also has the broadest altitudinal range (sea level to >6700 m [>22,000’]). To test hypotheses about adaptive regulatory responses to hypoxia, we will use a common-garden experimental design to integrate measures of whole-animal physiological performance (aerobic exercise capacity in hypoxia) and various subordinate traits (respiratory, cardiovascular, and metabolic) with tissue-specific transcriptomic and metabolomic profiles. Experiments will involve highly invasive manipulations (e.g., surgical instrumentation of arterial and venous catheters to measure blood gases during exercise trials and terminal sampling of vital organs) that are not feasible in human subjects. In both species, mechanistic experiments will be complemented by population genomic experiments to generate hypotheses about the specific genes and pathways that may have contributed to hypoxia adaptation. Such hypotheses will then be tested using follow-up experiments to measure phenotypic effects of changes in gene function and/or gene expression, as illustrated by our ongoing work on deer mice. The specific aims of the research are (1) to elucidate the mechanistic basis of adaptive enhancements of aerobic performance capacity in hypoxia; (2) to determine how regulatory changes in gene expression translate into changes in phenotype at different hierarchical levels of biological organization; and (3) to identify and experimentally test new candidate genes and pathways for hypoxia adaptation. The integration of population genomics, functional genomics, and experimental physiology will advance the field by elucidating the mechanistic basis of adaptive evolutionary change in complex performance traits.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Structured RNAs play fundamental roles in biological processes and are actively being pursued as targets to treat disease. Furthermore, synthetic RNA-based biosensors and therapeutics – inspired by natural RNA machines – are beginning to come online. These RNAs can undergo conformational transitions to bind small molecules and perform biochemical reactions. Unfortunately, incomplete models of RNA structure and how it recognizes molecules hinder the development of these potential novel devices and treatments. Without predictive biophysical models, the field relies on extensive experimental methods to probe RNA 3D structure, dynamics, and ligand binding. Experiments such as NMR, cryo-electron microscopy, phylogenetic analysis, and biochemical methods, although powerful, often fail to completely capture an RNA’s structural dynamics and conformational changes. To address these challenges, the Yesselman Lab is developing novel models of RNA 3D structure and design. We have demonstrated the first high-resolution RNA 3D helical thermodynamics model, automated design of RNA 3D structure, and developed novel experimental methods to probe secondary and 3D structures. Over the next five years, the Yesselman Lab aims to improve our understanding and predictive models of RNA conformational dynamics, RNA-ligand binding, and RNA catalysis: (1) RNA conformational dynamics: utilizing novel RNA 3D design and massively parallel biochemical assays, our goal is to develop general models for RNA 3D dynamics to understand better how RNA undergoes conformational transitions and binds to ligands. (2) RNA-ligand interactions: our goal is to build the first predictive model of RNA/drug interactions by assaying the effects of small molecule drugs on thousands of RNA structures combined with novel machine learning approaches. (3) RNA catalytic activity: self-cleaving ribozymes can cut RNA strands. Compared to proteins, ribozymes are significantly less active. A key difference is ribozymes often have less 3D scaffolding. When they do, they contain long-range tertiary contacts, but the strength and placement of these interactions can dramatically affect catalysis activity. Determining the rules of 3D scaffolding in ribozymes will increase our understanding of RNA catalysis and enable the design of new ribozymes for other catalytic functions. Findings from these research areas will address challenges and advance knowledge in RNA folding, molecular recognition, and design. Ultimately, our research program will play a critical role in developing the next generation of RNA-based diagnostics and therapeutics.

NIH Research Projects · FY 2025 · 2022-09

Antisense oligonucleotides (ASOs) are short (~20 bp) oligonucleotides that have chemically-modified backbones to resist endonuclease activity in cells and biological fluids. ASOs bind to targeted mRNAs within cells and act as catalysts for RNAse H to destroy the mRNA, thus decreasing gene expression. The most common ASO modification is the substitution of the unbridging oxygen atom of the phosphodiester group with a sulfur atom to create the phosphorothioate (PS) moiety which is most often used in clinical ASOs to date. Further stabilization and nuclease resistance may be conferred through the modification of the 2’ position of ribose of the ASO. It is widely known that the liver is the natural sink for PS-ASOs, however, the mechanism for this activity is not clear. We have discovered that the Stabilin class (SR-H) scavenger receptors (Stabilin-1 and Stabilin-2) are the primary mechanism for systemic PS-ASO clearance. Stabilins are expressed in a number of tissues including the sinusoids of liver, lymph nodes, spleen, Type II macrophages, bone marrow, etc which may have implications for PS-ASO delivery to many tissues. The gap in knowledge is how interactions of PS-ASOs with the Stabilins increases PS-ASO knock-down of targeted mRNAs. Our central hypothesis is that Stabilin-mediated endocytosis of PS-ASO proceeds along 2 pathways; one in which the PS- ASO is shuttled to the lysosome (destruction) and the other in which the PS-ASO is allowed to escape the endosome to interact with mRNAs (efficacy). Our primary objectives for this project are first, to determine the biological interactions of Stabilin-PS-ASO binding complexes with the use of biolayer interferometry with competing ligands in both known solutions and in plasma. Second, to determine the kinetics of PS-ASO endocytosis in both recombinant stable cells lines expressing the Stabilins and in primary sinusoidal endothelial cells of liver. We will also dissect the endocytosis mechanisms of Stabilin-2 in which we will elucidate interacting molecules that are necessary for PS-ASO activity (endosomal escape). Third, to determine the systemic clearance and bioactivity of PS-ASOs in WT and Stabilin knock-out mice to assess Stabilin-dependent biodistribution and activation in tissues other than just the liver. We will use global and tissue-specific Stabilin knockout mice for these studies. This will further delineate the two possible pathways (destruction vs activation) in multiple tissues and how the Stabilins contribute to each pathway. The expected outcomes of this project will lend greater understanding for the structure-activity relationship, efficacy, and overall metabolism of clinical-grade PS-ASOs and Stabilin biology/biochemistry.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Youth experiencing homelessness (YEH) face many significant risks that are detrimental to their long-term well- being and survival, including engaging in negative coping behaviors such as alcohol and drug use. 89% of YEH report lifetime alcohol use, 77% marijuana use, and 48% report methamphetamine use. Drug and alcohol use prevalence rates are 2 to 3 times higher among YEH than in their housed peers, leading to poorer mental health and HIV outcomes for this population. Proximal risk factors for substance use in YEH include street victimization: 39% and 94% have been sexually or physically victimized, respectively; poor mental health: 80% and 74% meet diagnostic criteria for PTSD and depression, respectively; and risky social networks: 62% of YEH report drug use with a network member. Another risk factor for substance is affect regulation. Studies of general population youth find that those with greater negative affect have higher rates of substance use. The role of affect regulation has rarely been examined for YEH; thus, mediating mechanisms are unclear. It is possible that proximal risk factors increase negative affect, leading to greater substance use among YEH. Understanding these linkages is key to developing predictive models of coping. This project uses a socio-ecological framework to examine linkages between Proximal risk factors, Affect regulation, and Coping behaviors. Specifically, we will identify factors that influence positive vs. negative coping behavior for YEH to develop a data-driven intervention that: 1) is relevant to this vulnerable population, and provides individualized support, and 2) can be sustainable outside the research setting. Our team has pioneered the use of cellphones for ecological momentary assessment (EMA) in public health research (Khan), and with YEH (Tyler and Olson). Increased cellphone ownership among YEH alongside growth in urban WiFi hotspot coverage together create a public health opportunity leveraged in this project's aims: 1) determine whether affect regulation mediates the relationship between YEHs’ proximal risk factors and coping behaviors, achieved by analyzing survey data and EMA data on Proximal risk factors, Affect regulation, and Coping behaviors collected via app-based responsive EMA (rEMA) in an observational cohort study of N=300 YEH over 60 days; 2) develop a data-driven app-based just-in-time personal support intervention (JIT-PSI) that delivers responsive messaging to mitigate negative coping in favor of positive coping. This aim is achieved by developing predictive models of coping behavior based on machine learning analyses of Aim 1’s survey and rEMA data; and conducting focus groups with N=30 YEH and meetings with N=15 agency staff to design intervention messaging content responsive to a range of coping behaviors; 3) test the feasibility and acceptability of the JIT-PSI, achieved by an observational cohort study of N=300 YEH over 60 days, using surveys, alongside cellphone based rEMA, and JIT intervention delivery. Leveraging growing number of public free WiFi hotspots and shrinking cellphone hardware costs, this project will yield significant public health impacts by developing a sustainable, JIT behavioral intervention for the prevention of drug use among YEH.

NIH Research Projects · FY 2024 · 2022-07

PROJECT SUMMARY Internalizing problems represent a growing public health concern, particularly across the transition to formal schooling, when children face new academic, social, emotional, and behavioral challenges. Despite the prevalence of internalizing problems during childhood, factors underlying their development are not well understood relative to externalizing problems. Thus, the overall objective of the present study is to investigate developmental pathways originating during pregnancy and unfolding across early childhood that contribute to school-age internalizing problems by leveraging an ongoing, large-scale, multi-method, longitudinal study of families. Based on prior theory and empirical evidence, it is expected that specific dimensions of parental trauma-related distress (TRD) spanning pregnancy to preschool will be associated with internalizing problems at age 5. Further, it is hypothesized that observed and self-reported emotion-related socialization behaviors (ERSBs) during preschool-age will mediate the link between parental TRD and school-age internalizing problems. Lastly, it is expected that nonsupportive reactions to children’s negative emotions, specifically, will demonstrate unique incremental prediction of internalizing problems beyond other forms of ERSBs, such as poor emotion coaching and supportive responses. Study aims will be pursued in an established community sample of 159 families (mothers, fathers, and children) who have completed self-report measures during pregnancy, at 1- and 6-months postpartum, and at child ages 1 and 2. Mothers, fathers, and children were invited to attend a laboratory appointment when the child turned 3.5, where parental ERSBs were assessed via self-report and observational coding paradigms. As part of the proposed project, mothers and fathers will be invited to complete a survey of child internalizing problems from home when the child is 5 years of age. Findings are expected to improve prevention and early intervention efforts for school-age children with internalizing problems by identifying malleable factors within the early family environment (e.g., parental psychopathology and parenting behaviors) that ultimately impact the development of internalizing problems. The goals of this study will be accomplished within the proposed research training program, which is aimed at helping the fellow develop expertise in family systems, observational coding, and statistical analyses involving multi-method, longitudinal data. The training plan includes completion of relevant courses, attendance in targeted workshops, individual supervision and mentorship by experts in the field, and preparation for an academic career in clinical psychology.

NIH Research Projects · FY 2025 · 2022-06

The objective of this Food and Drug Administration (FDA) Center for Veterinary Medicine (CVM) Laboratory Response Network (Vet-LIRN) Diagnostic Laboratory Program is to build capacity and infrastructure to enhance veterinary related sample analysis capabilities for the FDA through cooperation with Vet-LIRN network laboratories. Specifically, testing capacity may be required in the event of animal food or drug related illness or other large scale animal food or drug related emergency events that necessitate large scale testing. Additional funding will support investigation and surveillance assignments of contamination or adulteration in animal food or drugs. Testing capacity and capabilities enhanced at the University of Nebraska Veterinary Diagnostic Laboratory (UNLVDC) will include microbiological analysis and testing of samples such as animal food or drugs, environmental samples related to animal foods or drug production, and/or animal diagnostic necropsy or microbiological testing of clinical samples. In the event requiring testing for biological testing of food or drug products, the UNLVDC will perform selected analyses of diagnostic samples collected and supplied to the laboratory by FDA or other agencies as required. Although UNLVDC lacks a dedicated toxicology section, UNLVDC can contribute to the overall mission through support of microbiological and pathological testing as needed. The overall goal of the cooperative agreement is to support, utilize, and enhance university, state, and federal veterinary diagnostic laboratory testing capabilities during case investigations and to bolster capacity and support research to enhance the national food safety system. The program will also be used to provide analytical data using standardized methods, equipment, analytical worksheets, and electronic reporting. Demonstration of competency will include participation in proficiency testing and other interlaboratory comparisons and exercises as available. Standard quality management systems will be enhanced. Small scale method development and method validation projects will be performed, as directed by the Vet-LIRN Program Office. Funding from this agreement will be used for supplies, testing fees, equipment maintenance, travel for continuing education, and personnel time.

NIH Research Projects · FY 2025 · 2022-03

This project tests how various forms of social stress contribute to allostatic load and related health risk, and how aspects of social support networks may mitigate negative health effects of stress by reducing stress contagion. Our project focuses on two critical types of social stress: “day-to-day” stressful interactions and severe forms of social stress (e.g., experiences of violence), which may represent forms of stress resulting in acute and chronic changes to stress response, respectively. Changes in stress responding may contribute to worsening allostatic load, or the chronic “wear and tear” from stress that has consistently been linked to the development of chronic diseases such as diabetes and cardiovascular disease. This proposal will be among the first to test mechanisms of how social stress contributes to allostatic load; we will extend this work beyond the individual to understand how stress acts on a social network. Social support may improve several stress-related outcomes, including allostatic load, but data are lacking to explain how. Prior research does not account for how social support may perpetuate stress across a network: that is, when one person experiences stress, their supportive others will also experience stress. Put simply, when we experience social stress, we tend seek support from others with similar experiences, because they uniquely understand what we are going through: but when we do, we remind others of their own stressful experiences and stress “spills over” to them. However, some qualities of support networks may mitigate this spillover. The current study seeks to identify these qualities, which may be targeted in interventions that reduce stress not only in the individual, but across the social network. First, we will determine the extent to which social stress response and social stress exposure frequency combine to predict changes in allostatic load, and in turn predicts risk for the chronic health outcomes identified as priorities for the Presidential Make America Healthy Again agenda. Second, we will determine the social network qualities that mitigate stress “spilling over” from one person to another. Finally, we determine how social stress transmits across a network to predict changes in the networks’ allostatic load. To achieve this, we will use sampling designs (respondent-driven sampling; RDS) and social network analyses that allow us to understand how the connections across people influence health. We will examine social support; biomarkers of allostatic load, acute and chronic stress, and inflammation; and daily diary assessments of social stress frequency. Using novel Bluetooth technology developed by our team we will also assess how often participants interact with one another, linking that data to each participant’s reports of social stress and whether they discussed social stress experiences with supportive others. The combination of these measures and network-based approach allows us to test how social environments influence health. These data will inform how to best target social support interventions to limit the negative effects of stress.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT RNA is a structurally adaptive polymer that adopts complex tertiary structures and undergoes dramatic large- scale conformational changes to regulate cellular activity. Integral to RNA function is its remarkable plasticity and ability to structurally adapt in response to stimuli including small molecules and protein cofactors. Despite a central role in biology, a comprehensive understanding of the principles that govern RNA folding and molecular recognition is lacking. This research program addresses these gaps to understand, at an atomic level, how RNA folds and recognizes binding partners during normal cell function and in disease states. Using RNA aptamers and the 7SK ribonucleoprotein (RNP) as model systems, we combine solution NMR spectroscopy, X-ray crystallography, mass spectrometry, and basic biochemistry in a multidisciplinary approach to advance understanding of RNA structural dynamics and molecular assembly. The eukaryotic 7SK RNP is a major regulator of transcription. Comprised of the 7SK RNA and protein components, 7SK RNP binds and inactivates the kinase activity of the essential positive transcription elongation factor b (P-TEFb). P-TEFb must be released from the 7SK RNP to activate RNA Polymerase II transcription processive elongation. P-TEFb dysregulation or 7SK RNP malfunction is associated with several genetic diseases including cancers, heart disease, and primordial dwarfism. Moreover, several viruses manipulate host 7SK RNP for viral survival, notably HIV-1 and more recently SARS-CoV-2 underscoring the significance of 7SK RNP in biological processes. Despite its critical function and biomedical significance, there are presently few mechanistic insights into 7SK RNP function in stark contrast to other regulatory RNPs. This gap is largely due to a lack of fundamental knowledge on basic 7SK RNP features: how 7SK RNA folds, how proteins assemble onto 7SK RNA, and how 7SK RNP is structured. There is a critical need to answer these outstanding questions to provide foundational insights into 7SK RNP structural biology, and are essential to achieving a comprehensive understanding of 7SK RNP and its central role in biology. Over the next five years, we will determine high resolution structures of RNA aptamer-ligand complexes, 7SK RNA elements involved in P-TEFb release, and 7SK RNA-protein complexes. We will identify the determinants for RNA-ligand or RNA-protein binding specificity, elucidate 7SK RNA conformational dynamics, and uncover the 7SK RNP proteome and protein-protein interaction network during normal cell function and under genotoxic stress. Long-term, we will use newly gained knowledge to rationally design improved aptamer- ligand pairs, determine global folding and dynamics of 7SK RNA, identify the molecular mechanisms of P-TEFb release from 7SK RNP, and determine the 7SK RNP macromolecular architecture. Findings will address critical knowledge gaps in RNA molecular recognition, 7SK RNA structure, 7SK RNA-protein recognition, and RNP organization. This work will provide fundamental knowledge of RNA-protein interactions that can be extended to understanding the foundational principles of RNA-protein recognition for other RNPs.

NIH Research Projects · FY 2025 · 2021-08

Cellular metabolism is emerging as a critical factor to control the immune responses and their impact on the pathogens. In addition, recent studies pinpoint a more prominent role of the aberrant metabolism in controlling both genetic and epigenetic cellular phenomena of any form of cancer. Thus, investigating the dynamic metabolic shift in immune cells upon pathogenic infection and temporal ‘reactomics’ (defined as a combination of reaction mechanisms, regulations, and kinetic parameters) and associated vulnerabilities of tumor cells holds immense potential to develop novel therapeutic approaches. While the existing multi-scale modeling of immune cells tries to bridge the gap between multiple scales (i.e., molecular to organ-level), none of the existing approaches can simultaneously do that by building a proper, predictive ‘full-scale’ model. Furthermore, whether or to what extent metabolic shifts occur in the host’s immune system is still not known. In case of cancer cell, some of the critical challenges include defining the systems-level cellular metabolic phenotype and tracking the temporal changes in reactomics which are critical for reverting the cell metabolism to more healthy state. Herein, PI Saha proposes to develop and iteratively improve a systems-level, comprehensive, and integrative metabolic modeling framework: i) to dissect the dynamic shifts in the immunometabolic responses associated with pathogenic Infection, and ii) investigate the changes in temporal reactomics associated with the metabolic reprogramming in a specific cancer cell. The proposed research program will leverage the unique combination of computational modeling skills and rich research experience in Saha’s laboratory that are crucial for characterizing the metabolic phenomena associated with any disease. His research team recently developed the first computationally tractable and accurate modeling framework to track the temporal dynamics of cellular metabolism and also established a new method to estimate the reactomics of each of the metabolic reactions involved in a cellular system when ‘omics’ datasets are incomplete or missing and, thereby, develop a predictive kinetic modeling framework. Thus, the proposed modeling framework can potentially investigate the metabolic dynamics associated with a cluster of cells (e.g., immune cells) interacting with a pathogen or the temporal reactomics of a specific cell (e.g., cancer cell). As a first step, Saha will investigate the dynamic metabolic shifts in a specific type of immune cell (i.e., macrophage) upon SARS-Cov-2 and Staphylococcus aureus infection and the temporal reprogramming and reactomics of pancreatic ductal adenocarcinoma (PDAC) cell metabolism and test the hypothesis that if the degree to these changes gives rise to the severity of the disease symptoms. Overall, the proposed framework as well as the associated ‘predictome’ database (containing the predictions of key genes/proteins/reactions playing critical roles) will provide the broader scientific community including molecular biologists, computational biologists, clinicians, and translational scientists with a basic understanding of the role of metabolism in dictating disease severity and also a useful template to investigate other diseases.

NIH Research Projects · FY 2026 · 2021-08

PROJECT SUMMARY/ABSTRACT Endogenous peptides are naturally occurring transmitter molecules that act as messengers to facilitate communication between cells. Targeting endogenous peptide signaling is a proven strategy to treat a wide range of human diseases. A rigorous molecular-level understanding of how specific peptide molecules signal provides valuable insights into the cause of disease and directly informs the design of therapeutic compounds. However, there remain many bioactive and disease-relevant endogenous peptides whose signaling pathways are not well understood. Because of the importance of peptide-receptor interactions in both normal physiology and disease, there is a critical need for new tools and approaches to fully interrogate and modulate these signaling systems. The long-term goal of this research program is to develop and implement chemical approaches to build our understanding of peptide transmitters in biological processes and to identify new therapeutic targets. To achieve this goal, this research program pursues a highly interdisciplinary approach, with expertise in the design, synthesis, and application of novel chemical probes, protein and peptide mass spectrometry, and analysis of peptide-receptor interactions on cells. Over the next five years, this MIRA research program will focus on two complementary research areas. Research Area 1 of this program will develop and implement unbiased chemical approaches to directly detect peptide-receptor interactions on the surface of cells through affinity-driven protein labeling. These approaches will not require genetic or chemical modification of receptors prior to the labeling event and will be compatible with membrane proteins of low abundance. We will utilize these approaches to identify the receptors for physiologically relevant bioactive peptides, which will serve as novel therapeutic targets. Research Area 2 will develop and utilize mass spectrometry-based peptidomics methods to identify and quantify changes in cell-cell signaling peptides to understand general principles underlying natural adaptations for cellular protection. These methods will allow for the rigorous characterization of endogenous peptides extracted from tissues, including peptide quantitation with identification of all post-translational modifications. Peptides of interest identified from peptidomics will then be subjected to functional analysis to validate their role in physiology, enhancing our knowledge of these biological processes and identifying new targets for therapeutic modulation. The overall vision of this MIRA research program is to develop new chemical strategies to fully understand and modulate peptide signaling pathways. This work will impact biomedical research by enhancing our knowledge of the molecular mechanisms of endogenous cell-cell signaling peptides, building new chemical tools to study these signaling events, and identifying new therapeutic targets.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT Nitric oxide (NO) is a key signaling molecule in biological systems, but it is poorly understood due to a lack of temporal and spatial sensors that measure NO. A direct, fast and accurate sensor for measuring NO would be of immense value to ongoing and future research. Single walled carbon nanotubes (SWNT) have excellent potential for use as NO sensors both in vitro and in vivo, but they are not widely used by the research community because they are not readily available in an easy-to-use platform. The goals of this project are to 1) develop new SWNT sensor platforms and modify the current systems to improve sensor specificity and handling, and 2) determine nitric oxide concentration and dynamics with a highly sensitive and accurate sensor. The goals will be accomplished through the completion of four specific projects. In the first project we will develop a ratiometric sensor to quantify NO over long time periods, the second project will involve the development of an easy-to-use platform for the in vivo delivery and stabilization of SWNT sensors that are capable of both spatial and temporal analyte quantification. Projects 3 and 4 will use the SWNT sensors to investigate NO concentrations, specifically for project 3 we will determine the intracellular dynamics of NO in relation to organelles and membranes and project 4 will involve the investigation of extracellular NO concentration values and gradients associated with healthy and diseased cells. This work will leverage the expertise of the Iverson Laboratory to engineer new and improved sensor delivery platforms and use the sensors to investigate intracellular and extracellular NO signaling. By developing these platforms a template for the development of other SWNT sensor systems will also be provided, allowing researchers to learn about reactive oxygen species, small molecules and proteins in a spatial and temporal fashion at the sub-cellular level. The investigation of the NO signaling for the cells investigated in this project will provide researchers with a basic understanding of NO’s role in cell signaling and provide a template for investigation of other cell types.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY Iron-sulfur (Fe-S) clusters are ancient cofactors composed of multiple iron and sulfur atoms. They are fundamental to numerous biological processes in all domains of life. Owing to the rich, tunable redox reactivity and selectivity of the cluster, Fe-S proteins play multifaced roles in redox control under both physiological and stress conditions. The roles of Fe-S proteins in redox control are vital for the maintenance of normal cellular functions and cell survival, and thus they are tightly linked to health and disease such as cancer and diabetes in human and bacterial infection. This contrasts vividly with the lack of functional, structural and mechanistic understanding of many Fe-S proteins in the cellular control of redox homeostasis, particularly in the three core aspects that are addressed in this MIRA proposal: i) redox sensing and transcriptional regulation by Fe-S proteins; ii) assembly, transfer and repair of Fe-S clusters; and iii) crosstalk between Fe-S proteins and the low-molecular- weight (LMW) thiols in redox hemostasis. Several Fe-S proteins-mediated mechanisms for redox control in mycobacteria will be used as examples to elaborate our research goals and approaches in this proposal, including i) redox sensing and transcriptional regulation by a unique family of Fe-S cluster-bound transcription factors in the WhiB-like family; ii) assembly, transfer and repair of Fe-S clusters by the SUF system; and iii) mycothiol in Fe-S cluster homeostasis. The proposed research program is built on the unique combination of skills and rich research experience in my research team that are crucial for characterizing the oxygen-sensitive metal-binding proteins. We recently determined the first and long-desired Fe-S cluster-bound structure of the monomeric transcription factor WhilB1 from Mycobacterium tuberculosis and established a new mechanism of bacterial transcriptional regulation mediated by this protein. By combining structural, spectroscopic and biochemical approaches in vitro with molecular biology in vivo, we are poised to determine: i) the structural basis of redox reactivity and ligand selectivity in Fe-S clusters; ii) the mechanism by which the redox state and integrity of the Fe-S cluster allows these proteins to sense redox state and regulate transcription; iii) the structural biochemistry of Fe-S cluster biosynthesis and regulation; and iv) the role of non-proteinaceous thiols in modulating Fe-S cluster-mediated redox control. Altogether, the proposed research program will establish a new line of ground-breaking research in an understudied aspect of redox homeostasis. The strategies developed from the proposed program will be instrumental for studies on the newly discovered Fe-S cluster system, such as those in the DNA repair and RNA metabolism in response to oxidative stress. Because of their critical roles in redox control, the study of the novel mechanisms of these Fe-S proteins will not only shed light on the fundamental molecular mechanism governing Fe-S protein-mediated redox control, but also have a significant impact on improving health and combating infectious diseases.

NIH Research Projects · FY 2025 · 2020-07

Project Summary This proposal seeks funding to continue support for the Molecular Mechanisms of Disease (MMoD) predoctoral training program at the University of Nebraska-Lincoln. The MMoD program addresses the NIH training focus of Cellular, Biochemical, and Molecular Sciences by providing advanced training and career development in biomedical research centered on molecular interactions and chemical transformations. The goal of the program is to develop outstanding new scientists who work in collaborative multi-disciplinary teams to research disease mechanisms using quantitative approaches that ultimately yield tangible strategies for prevention and therapy. To accomplish this goal, the program recruits high-quality, motivated predoctoral trainees with a strong interest in the underlying causes of human disease and engage them in mentored, cutting-edge research. Program outcomes from the last four years indicate strong progress toward our goal with 17 appointed trainees thus far. The MMoD program provides collaborative training in mechanistic disease research in the areas of molecular signaling, metabolic integrity, oxidative stress and redox biology, and disease microenvironment. The mentoring team spans seven departments and three colleges and includes 32 faculty members from early-stage to established investigators with strong histories in biomedical research funding and graduate student mentoring. The MMoD program will (1) Provide a rigorous curriculum and innovative, collaborative research opportunities for six predoctoral trainees per year (NIH-supported) to become experts in the mechanistic study of human disease; (2) Cultivate an interactive and inclusive training environment that enables trainees to develop professional skills in communication, leadership, proposal-writing, scholarship, entrepreneurship, and teaching; and 3) Build a cohort of trainees from diverse perspectives that have the knowledge and ability to work at the crossroads of different disciplines in their future careers as PhD scientists. Trainees will conduct research rotations in year 1 before choosing a faculty mentor, and will be supported by NIH T32 funding for up to two years beginning in their second year of their five-year program. Selection of new trainees for NIH support will be competitive and includes an original collaborative proposal bridging disciplinary boundaries for innovative thesis research. Trainees will take a flexible core curriculum that emphasizes writing and quantitative biology skills. Students in the MMoD program acquire skills in evidence-based education, quantitative and critical analysis, and rigor and reproducibility in research. They also gain professional experiences related to intellectual property and the pharmaceutical industry in preparation for potential career options. The MMoD program provides a framework that enables trainees to gain a broad knowledge base; actively seek research collaborations; produce an outstanding record of original published research; and develop presentation, proposal-writing, and leadership skills that position them for future excellence as independent researchers.

NIH Research Projects · FY 2026 · 2019-06

PROJECT SUMMARY/ABSTRACT Mitochondria are complex, dynamic organelles that are essential in virtually all eukaryotes. They play important roles in various vital processes including energy conversion, synthesis of iron-containing cofactors and metabolic intermediates, immune signaling, and cell fate choices. Although knowledge about mitochondrial biology is increasing, the mechanistic understanding of how mitochondrial functions are established, maintained, and adjusted in response to physiological and stress cues remains incomplete. This is significant as said mechanisms are highly relevant to both normal cellular physiology and a wide range of Mendelian and common age-related human disorders – including glaucoma, hearing loss, Parkinsonism, amyotrophic lateral sclerosis, various neuropathies, dementia, and certain cancers – for which no effective therapies currently exist. The overarching goal of this research program is to determine how evolutionary conserved mitochondrial quality control modules function to maintain cell survival and how these functions can be manipulated to achieve clinical benefits. Unified by the topic of mitochondrial fidelity and protein homeostasis, the program utilizes multidisciplinary approaches to address key gaps in knowledge about conserved mechanisms through which compartmentalized protein folding, stability, assembly, and membrane integrity are safeguarded to ensure proper mitochondrial functions. The proposed studies focus on the following directions: 1) Understanding disease-relevant mechanisms that govern proteolytic and proteostatic control at the inner mitochondrial membrane; and 2) Elucidating how a delicate protein homeostasis in the mitochondrial matrix is established and maintained. In each direction, the proposed work will focus on a group of dedicated factors that mediate protein, ion, and metabolic homeostasis in mitochondrial subcompartments, thereby ensuring the organelle's self-preservation under basal and stress conditions. Capitalizing on previous findings and novel tools and approaches developed by the PI and colleagues, the team will identify and characterize the mechanisms behind these processes. Expected outcomes will deepen the fundamental understanding of mitochondrial biology and related pathogenic mechanisms of human diseases stemming from progressive mitochondrial dysfunction due to quality control of failing organelles.

NIH Research Projects · FY 2026 · 2019-04

Treatment and prevention of women’s substance use lags that of men, and interventions designed for men are often ineffective for women. Much prior work has failed to account for unique contextual factors related to young women’s substance use: relative to men, women report greater stress-induced and cue-induced substance cravings and are more likely to progress from first use to clinically significant substance use in a short period of time. Also, about 1/5 – 1/3 of women report using substances for sexual enhancement (i.e., sex-linked substance use; SLSU). When women frequently pair substance use with sex, sexual desire may become a substance-related cue that triggers cravings. Finally, there is high co-morbidity of substance use disorder with mood and anxiety disorders, which are more prevalent in women. Mood symptoms reinforce substance use and vice versa, and hence successful intervention requires knowing which symptoms to target to cause a beneficial “chain reaction” that disrupts the broader symptom network. The long-term goal of the proposed work is to prevent SUD in young women at a pivotal developmental window that sets the stage for lifelong substance use patterns. The proposed project’s objective is to identify potential tractable points of intervention to effectively disrupt women’s substance cravings and escalating substance use trajectories. The central hypothesis is that women’s substance cravings and stress interact to reinforce the connections between SLSU and mood symptoms, which leads to rapid increases in substance use. To test this hypothesis, a sample of young women will be recruited to assess networks of mood symptoms and substance use, including SLSU and cravings. Repeated survey measures will be used to estimate within-person symptom networks with much higher accuracy and precision than traditional single-timepoint designs. The two aims of this project are to identify: (1) the dynamic role of stress in changes in women’s substance cravings over time, and (2) the factors most central to change in the network of women’s substance use and mood symptoms. Data from this study will identify how stress and sexual/relational contexts interact to trigger women’s substance cravings, how changes in symptom network stability predict women’s substance use trajectories, and critical factors to tailor substance use prevention efforts to women. These data will support Project Leader Tierney Lorenz in submitting an R01 to test prevention-based interventions in young women who use substances. Ultimately, this will contribute to the development of effective programs to slow women’s substance use trajectories and prevent development of SUD, a critical area of progress within the Rural Drug Addiction Research Center’s wider aims.

NIH Research Projects · FY 2025 · 2018-07

Project Summary Obesity is a prevalent and impairing condition that poses a major public health challenge in the United States. To face this challenge, there is a need for ground-breaking research aimed at identifying modifiable factors that contribute to obesity risk at pivotal junctures, thus informing novel interventions. Studies aimed at elucidating specific neural vulnerability factors that predict future excess weight gain during the critical period of adolescence hold particular promise. Findings from the parent R01 point to the potential role of top-down regulation in emerging obesity risk in adolescence. Top-down regulation deficits are associated with higher body mass index (BMI) and more obesogenic eating behaviors. While these results suggest the importance of top-down regulation in emerging obesity risk, critical questions remain and present opportunities for novel investigation, including the relative impact of food-specific versus non-food regulation, the impact of regulation within the context of reward sensitivity, and the interactions between neural vulnerabilities and food environment in predicting adolescent weight trajectories. Guided by our Contextualized Neural Vulnerabilities Model of Obesity, the proposed renewal addresses each of these issues within a highly contextualized longitudinal study exploring regulation, reward, and environment – and the interplay between these factors – in predicting adolescent weight trajectories. We propose to follow a diverse sample of adolescents for a longitudinal multimethod study leveraging rigorous measures of neural vulnerability factors (measured via fMRI tasks), diet (via multiple 24-hour recall), weight status (via BMI and body fat %), and obesogenic food environment (via multidimensional assessment, including geocoded risk score). The long-term goal is to inform specific targets of novel interventions to reduce long-term obesity risk. The objective of the proposed research, therefore, is to elucidate the role of theoretically relevant neural vulnerability factors for obesity in context within a longitudinal study. The central hypothesis is that food- specific regulation plays a uniquely critical role in adolescent diet and weight trajectories, particularly in the context of high reward sensitivity and obesogenic food environment. The specific aims are to: 1) Determine the unique relative impacts of food-specific versus non-food regulation on adolescent diet and weight trajectories; 2) Determine the impact of food-specific regulation on adolescent diet and weight trajectories, in the context of reward sensitivity; and 3) Explore the interaction between neural vulnerabilities factors (regulation, reward) and obesogenic food environment in predicting adolescent diet and weight trajectories. The study is innovative in its conceptual framework, novel focus on neural vulnerabilities in context, and integration of diverse data collection modalities within a developmentally informed longitudinal design. The significance of this research is that it will directly inform the specific targets of interventions as well as the individual and environmental contexts in which these targets may be most relevant. These results will contribute to more personalized prevention approaches, with the potential to substantially strengthen obesity prevention efforts during a key developmental period.

NIH Research Projects · FY 2026 · 2016-09

PROJECT SUMMARY/ABSTRACT The immune system is arguably the second most complex human system after the brain. Its proper response to foreign stimuli is governed by network-like interactions among various types of cells and cytokines as their communication mediators. The complexity at the inter-cellular level of the immune system is further exacerbated by the similarly complex biological and biochemical networks within each cell (metabolism, gene regulation, etc.) responsible for the dynamics and decision-making at the single-cell level. Such multiscale complexity makes it incredibly challenging to understand the complete etiology and pathology of immune-system-related diseases. My research program aims to identify how the immune system can be rewired en masse to elicit higher-order decision-making while still enabling the system to remain otherwise “healthy.” To this end, my research program is leveraging a highly interdisciplinary research team (computational and experimental immunologists, software engineers, and education researchers) and collaborators to build a Virtual Immune System -- a multi-scale, multi-approach computational framework to understand better the complex dynamical nature of the immune system, identify more accurate multi-dimensional biomarkers, and identify safe and effective treatments within a reasonable time and cost. In the next five years, in addition to expanding the Virtual Immune system, my program will continue to develop methods and technologies for data-driven model construction, visualization, computation, real-time simulations, and reproducibility to advance multi-scale modeling of the immune system and beyond. We will continue to decipher the dynamics of the immune system under various pathologies and the re-programmability of CD4+ T cells under the milieu of their microenvironments. My laboratory will continue to iteratively predict, validate, and refine predictions generated using the systems approaches and technologies. To do this, we will generate our multi-omics data to more precisely validate immune system behaviors and apply our findings to refine the computational approaches directly. My team will continue to build collaborations and deepen our existing relationships, including with translational partners to advance the impact of our systems work on drug discovery, the international team modeling COVID-19, and with virologists and immunologists to further validate our computational predictions experimentally.