Ohio State University

NIH Research Projects · FY 2025 · 2024-09

Project Abstract While obesity has been shown to increase mortality in sepsis, the mechanism underlying this phenomenon is unclear. Dr. Jalilvand’s preliminary data demonstrates that obesity is associated with increased 90-day mortality in surgical patients with sepsis. Further, we have shown that obese adipocytes produce distinct adipocyte- derived extracellular vesicles (AdEVs) that activate neutrophils and induce disordered neutrophil swarming. These AdEVs are enriched with miR-223, which interacts with PIK3C2A to downregulate the phosphoinositide- 3 kinase (PI3K) pathway, thus reudcing neutrophil migration and chemotaxis. The governing hypothesis for this proposal is that AdEV-mediated delivery of miR-223 to peripheral neutrophils in obesity impairs neutrophil swarming by modulating the PI3K pathway, thereby contributing to disrupted pathogen clearance in surgical patients with sepsis. We will test this hypothesis with the following aims. AIM 1: Define the role of AdEV-mediated delivery of miR-223 on PI3K activation and swarming in healthy neutrophils and the efficacy of targeting this pathway to restore swarming in septic obese and lean patients in vitro. We will determine the in vitro differential impact of modulating AdEV miR-223 content on neutrophil function and whether pharmacologic upregulation of the PI3K pathway can restore swarming in neutrophils isolated from healthy and septic obese and lean individuals. AIM 2: Establish the efficacy of inhibiting miR-223 mediated PI3K downregulation in neutrophils to improve swarming and survival in obese mice with intra-abdominal sepsis. Using a murine model of diet-induced obesity (DIO) and intra-abdominal sepsis (cecal ligation puncture, CLP), we will test whether systemic delivery of AdEVs enriched with anti-miR-223 improves neutrophil function, bacterial clearance, and survival following CLP in vivo. Lastly, using a conditional, neutrophil-specific knock out of PIK3C2A, we will establish whether transient silencing of PIK3C2A mitigates miR-223-mediated downregulation of neutrophil PI3K activation and swarming in DIO mice after CLP. While capitalizing on her PhD in adipose tissue inflammation and obesity, this proposal represents a departure from both Dr. Jalilvand and her primary mentor’s, Dr. Hsueh, prior research focus. A multidisciplinary approach has been crafted to study the proposed aims to develop Dr. Jalilvand’s expertise in EV biology and cargo, live cell imaging to follow neutrophil behavior and quantitative function, pharmacologic targeting, and murine models of sepsis. Completion of this proposal will provide crucial technique expansion needed to establish a unique area of research distinct from Dr. Jalilvand’s PhD, accomplishing the short-term goals of this career development plan. Paired with the institutional commitment to her success that includes laboratory space, protected time, and laboratory funding, a well-established committee of mentors has been selected to ensure that Dr. Jalilvand will achieve the long-term goal of becoming an independent surgeon scientist evaluating the implications of obesity in surgical patients with sepsis. As such, this proposal will provide Dr. Jalilvand with the requisite mentorship and resources to transition to independence.

NIH Research Projects · FY 2025 · 2024-09

Abstract Acquired infections after spinal cord injury (SCI) constitute the main cause of death in patients. For those that survive, acute pneumonia has been associated with significantly reduced neurological recovery. However, proof of causality and the mechanisms underlying the blunted recovery triggered by infections remain elusive. To analyze whether pneumonia has a causal and direct effect on the lesioned spinal cord, we expose mice to a clinically relevant, controlled SCI-associated pneumonia (SCI-AP) embedded into a bed-to-bench translational approach. Our experimental data demonstrate that contracting pneumonia after subacute SCI with consecutive intraparenchymal iron deposition extending from the lesion site into spared tissue. Three aims are proposed to answer one main and novel question: does acute pneumonia exacerbate spinal cord -AP-triggered signaling as causal targets for neuroprotective intervention. Experiments in Aim 1 apply parabiosis models in conjunction with serum injections to identity circulatory, humoral factors underlying SCI-AP-induced injury/disrepair. Moreover, tracing bacterial fragments will detect passive shuttling of immunogenic bacterial material (Bright fluorescent S. pneumoniae) across a breached blood spinal cord barrier to study possible direct spinal cord lesion-pathogen interactions. Loss of vascular integrity will be detailed to the capillary level to detect features of capillary fragmentation by using spinal cord tissue clearing, intravascular tracing and 3-D imaging. Aim 2 targets and its underlying IFN-1 signaling via toxic CD11b+, CD206-- to propagate vascular damage and/or disrepair (STINGflox x LysMcre mice). Aim 3 captures SCI-AP-induced cell-specific transcriptional changes associated with anti-microbial immunity, such as myeloid cell-dependent effects on spinal cord endothelium to reveal novel targets for blocking or reversing SCI-AP-induced injury/disrepair. Clinically, SCI-AP typically emerges around day 2-3 days post SCI when the patient is in the hospital. This offers a more feasible and realistic time window for protective interventions compared to orthodox neuroprotective strategies which are limited to a short post- SCI time-window (golden first hour(s)). If successful, data from these experiments will directly inform effective strategies for SCI patients to i) reduce infection-associated disability and ii) protect outcome at risk.

NIH Research Projects · FY 2024 · 2024-09

ABSTRACT Ependymal motile cilia are responsible for the coordinated movement of cerebrospinal fluid (CSF) along the ventricle wall. The synchronized activity of ependymal cilia creates specific flow networks that are regulated by behavioral state. Ependymal cilia motility can be controlled by specific compounds in the CSF to adjust fluid flow, but little is known about the molecular and cellular mechanisms of this regulation. We propose the hypothesis that G protein-coupled receptor (GPCR) signaling pathways are expressed and function within ependymal cilia to modulate ciliary beat frequency. The focus of this proposal is to define the complement of signaling receptors that localize to ependymal cilia and determine how GPCR activation impacts cilia beating and activity-dependent ciliary localization of regulatory signaling proteins. As proof of principle, the GPCR melanin-concentrating hormone (MCH) receptor 1 localizes to ependymal motile cilia and we find that the GPCR signaling protein β-arrestin moves into ependymal cilia in response to MCH treatment. To assess which other GPCRs and signaling proteins are localized to motile cilia in ependymal cells, we will perform proximity labeling in normal and Bardet-Biedl syndrome protein deficient ependymal cells to identify low abundance and transient ciliary signaling proteins. We will also employ pharmacological approaches to evaluate the role of potential ciliary GPCRs in the regulation of ependymal cilia beating. Finally, we will assess potential ciliary GPCR activation and signaling by defining the spatial organization of GPCRs, β-arrestin, and the signaling chaperone Bbs4 within and proximal to the ciliary compartment prior to and after GPCR agonist addition. In order to provide the necessary resolution to define the precise spatial organization, we will use expansion microscopy, which is a cutting-edge imaging technique. The outcomes of this work will provide critical insight into the sensory and signaling functions of ependymal motile cilia and lay the foundation to create new tools to control ependymal cilia activity and discern the role of ependymal cilia in the mature brain.

NIH Research Projects · FY 2024 · 2024-09

Project Summary Acute kidney injury (AKI) is a common and severe complication in hospitalized patients, which is associated with high morbidity and mortality rates. Patients who survive an episode of AKI are at increased risk for progression to chronic kidney disease and end-stage renal disease. Therefore, there is a dire need to identify therapeutic strategies to prevent or treat AKI. The kidneys have a high metabolic rate and substantial decreases in the levels of nicotinamide adenine dinucleotide (NAD+) and NAD+/NADH imbalance impairs energy generation and kidney function during AKI. Augmentation of NAD+ may protect kidney tubule against diverse stressors. However, how NAD+/NADH imbalance contributes to development of AKI remains unknown. Our preliminary studies involving a druggable genome screen have discovered an unexpected role of a transcriptional repressor CtBP2 (C-Terminal Binding Protein 2) in renal epithelial cell death. CtBP2 is a NAD+/NADH-dependent transcriptional repressor. Our studies show that CtBP2 is activated in ischemic, nephrotoxic, and rhabdomyolysis-associated AKI. Moreover, in multiple mouse models of AKI, pharmacological inhibition of CtBP2 mitigated renal impairment, suggesting a pathogenic role. Furthermore, we found that CtBP2 is acetylated and this is critical for nuclear localization during AKI. Our ChIP-seq (chromatin immunoprecipitation followed by sequencing) studies showed that CtBP2 transcriptionally suppresses many genes involved in energy metabolism, including NAD+ biosynthesis in AKI. We hypothesize that CtBP2 is a pathogenic regulator of RTEC dysfunction and cell death that regulates NAD+/NADH imbalance-mediated metabolic dysregulation as a stress-induced metabolic sensor during AKI. To test this hypothesis, we will investigate a role of p300 in nuclear localization and activation of CtBP2 in AKI (Aim 1) and determine a pathogenic role of CtBP2 in renal injury and metabolic changes during AKI (Aim 2). These studies will significantly advance our understanding of the molecular mechanisms of CtBP2 activation during AKI. These outcomes will define novel molecular networks that underlie the severity of AKI and could lead to development of CtBP2 inhibition as a novel therapeutic strategy.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Over 1 billion people around the world will be significantly impacted by a neurological disorder during their lifetime. Better understanding of the underlying pathobiological mechanisms and the development of new treatment paradigms is critical to mitigate the impact and/or cure these diseases. Neurosurgeon-scientists have a unique opportunity to fundamentally advance understanding into ineffectively treated or untreatable neurologic disorders. Specifically, neurosurgeons can directly access human physiologic data, acquire nervous system lesions/tissue, develop/employ new surgical treatment technology, as well as make direct associations between the operating room and clinical findings. Moreover, they have the potential to collaborate with scientists from other disciplines to most effectively advance critical discoveries and develop new therapeutic paradigms. Consequently, defined and robust training of neurosurgeon-scientists is urgently needed. To enhance research training and best support the successful transition from residency to an independent neurosurgeon-scientist career, we are applying for National Institute of Neurological Disorders and Stroke (NINDS) R25 in Neurological Surgery at the Ohio State University (OSU). The overarching objective of this program is to provide research training to neurosurgery residents in the basic, translational and/or clinic sciences that will drive their successful transition to independent National Institutes of Health (NIH) funded neurosurgeon-scientists after residency. This program is led by 2 senior investigators (Drs. Herson and Lonser) who have implemented a research curriculum and programmatic changes to maximize the impact of the R25 on selected neurosurgery residents at OSU. The training plan is supported by senior faculty with a track record of mentoring/funding success and leverages the research infrastructure across the OSU campus.

NIH Research Projects · FY 2025 · 2024-09

Reading difficulties continue to be a pervasive public health issue. Many children experience reading difficulties due to underdeveloped phonological awareness – and, specifically, phonemic awareness. Although decades of basic research have established phonemic awareness as causally related to reading acquisition as well as a developmental progression from phonological sensitivity to phonemic awareness, we know much less about how or when to support phonological awareness development in children. Our long-term goal is to optimize phonological awareness intervention as a means of preventing reading difficulties. Our short-term goal is to complete two randomized controlled trials that inform such optimization through addressing current scientific controversies around the content, timing, and goals of phonological awareness intervention. In the randomized controlled trials, we evaluate three interventions (phonological sensitivity + phonemic awareness, phonemic awareness only, delayed phonemic awareness), which vary in the linguistic units targeted but are carefully equated on content and/or instructional time, for their impact on children’s phonemic awareness, reading, and spelling skills relative to each other and to a control condition; we also consider whether impacts differ based on when and to whom intervention is provided. Participating preschool and kindergarten children contribute screening, pretest, midtest, posttest, and longitudinal data to address four aims: (Specific Aim 1) Determine the relative impact on phonemic awareness, and subsequent reading and spelling development, of intervention that (a) initially targets phonological sensitivity or (b) directly targets only phonemic awareness, (Specific Aim 2) Determine the relative efficacy of these two approaches when intervening during preschool or kindergarten, (Secondary Aim 3) Examine whether the impact of the two approaches is differential based on children’s initial phonological skills and alphabet knowledge as well as sociodemographic factors, and (Secondary Aim 4) Explore whether a transition point can be identified as to when phonemic awareness intervention is most likely to be efficacious. The results will contribute to both basic science concerning the development of phonological awareness and its contribution to reading and spelling acquisition as well as translational efforts. With respect to the latter, results will inform learning standards, instructional recommendations, and intervention design to better align these with the scientific evidence base and thereby improve prevention and intervention for reading difficulties.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Each year, ~1 million Americans survive a critical illness only to suffer with new-onset frailty. This accelerated, catastrophic, critical illness-associated form of frailty is an emerging, costly, and age-related public health problem that is driven by the growing number of survivors of critical illness, the aging U.S. population, and the ongoing COVID-19 pandemic. Little is known about critical illness-associated frailty. The lack of systematic knowledge of critical illness-associated frailty makes it difficult to propose and test potential interventions. We propose to validate questionnaire-based frailty assessment methods needed to identify those with baseline (i.e., pre-critical illness) frailty, leading directly to a new generation of interventions to manage and mitigate critical illness-associated frailty, ultimately improving the long-term health for older adults surviving critical illness. We hypothesize that questionnaire-based frailty assessments will demonstrate strong reliability and validity as compared to performance-based methods. Upon completion of this study, we will have validated questionnaire-based frailty assessments that can be used when performance-based frailty assessments are not possible—an important next step in our work to understand and intervene upon critical illness-associated frailty. Aim 1: Determine the validity and reliability of patient responses on questionnaire-based frailty assessments with performance-based frailty assessments. Aim 2: Determine the validity and reliability of surrogate responses on questionnaire-based frailty assessments with performance-based frailty assessments. The study of critical illness-associated frailty is in its infancy; our project is therefore intrinsically innovative. Additional innovation arises from our systematic and rigorous frailty measurements using performance-based assessments and questionnaire responses from both patients and their carefully selected surrogates. This is the first study to conduct systematic measurement of questionnaire-based frailty assessments in relation to performance-based frailty assessments in those with critical illness. We will address key barriers needed to guide our approach to intervention, giving this project broad significance. To date, pre-critical illness frailty has been measured by judgement-based tools. Rigorous and objective measurements of frailty are needed to understand etiologies, risk factors, and to guide interventions. Regardless of outcome, results will improve the health of millions of older adult survivors of critical illness by identifying the extent to frailty is present at ICU admission facilitating the study of trajectories, risk factors, and mechanisms of a common, costly, and preventable condition—critical illness-associated frailty.

NIH Research Projects · FY 2025 · 2024-09

Healthy lifestyle behaviors, including diet and physical activity, reduce cancer risk and mortality. Although dietary guidelines have shifted to focus on dietary patterns, evaluations of dietary patterns have primarily relied on self-reported data. Self-reported dietary intake has systematic measurement errors and could lead to incorrect dietary pattern estimation. Dietary biomarkers provide unique opportunities to improve the reliability of self-reported dietary patterns. Given the current focus on recommending healthy dietary patterns, developing a method for calibrating self-reported dietary patterns to mitigate measurement error and to strengthen the existing evidence of diet-cancer association is necessary. Additionally, meal frequency and timing are critical aspects of nutrition and health. Metabolic aberrations are a cancer risk factor. Understanding the role of meal frequency on metabolic biomarkers and appetite regulation can provide new insights to inform the design of dietary interventions to improve metabolic health and reduce cancer risk. Lifestyle programs, such as consuming a better diet and engaging in physical activity, could reduce cancer recurrence and improve overall survival. However, whether lifestyle interventions delivered through various approaches lead to different physiological responses among cancer survivors has not been established. It is possible that due to personal and environmental differences, each cancer survivor may benefit differently from various delivery approaches. The proposed application focuses on: a novel approach for dietary assessment, physiological mechanisms of energy balance and cancer, intervention modalities to deliver lifestyle modifications, and intervention strategies to effect change in all populations. These objectives align with Make America Healthy goals to improve diet and physical activity across all populations for cancer prevention. The successful completion of the activities in this award will strengthen the foundation necessary to launch my career and position me with the expertise to become an independent investigator with a focus on lifestyle modifications in cancer prevention and control to reach all populations.

NIH Research Projects · FY 2025 · 2024-09

Gynecologic Cancer remains an underfunded area of research with funding to lethality scores significantly lower than other cancer sites. Endometrial cancer, the most common gynecologic cancer, is one of the few cancers that has seen an increase in incidence and mortality over the recent years.. High risk and rare subtypes of uterine cancers have distinct molecular profiles. Recent research has identified several molecular subtypes that can be targeted with novel agents. Molecularly driven trials are critical to the progress of personalized medicine to improve survival and limit exposure to ineffective therapy and associated toxicities. Unfortunately, these highly specialized clinical trials are at risk of failure due to slow accrual due to the rarity of certain cancer subtypes. As the national PI for NCI/NRG-GY020 (vaginal brachytherapy +/- immunotherapy for early stage high risk mismatch repair deficient endometrial cancer, which is a subset of endometrial cancer patients who are highly sensitive to immunotherapy) Dr. Backes is well aware of the opportunities and challenges associated with molecularly driven clinical trials. NCI support through the R50 mechanism will allow Dr. Backes to increase her efforts to improve enrollment to clinical trials for all populations, both at her institution and nationally. First, leveraging the resources of the Ohio State University Comprehensive Cancer Center (OSUCCC), Dr. Backes will develop a comprehensive system for molecular matching to simplify and streamline the identification of patients for trials and specialized trials for patients, which will increase accrual. Collaborative cross-disciplinary efforts are especially important with the increasing complexity of molecularly driven clinical trials, such as NCI ComboMatch and iMatch, and also to improve the outcomes of patients with rare tumor mutations. Secondly, Dr. Backes will collaborate with Dr. Elektra Paskett to include gynecologic oncology on a recently opened study that aims to connect patients to clinical trials. Dr. Backes and Dr. Ashley Felix (PhD, Epidemiology) will focus on assessing barriers and implement strategies to improve access to care and clinical trials in gynecologic oncology locally and nationally. Finally, the opportunities afforded by this award will allow Dr. Backes to increase her time to design and support the development of paradigm shifting clinical trials, and mentor junior investigators in this process, through her national roles as the NRG Oncology Co-Chair for Developmental Therapeutics, member of the Uterine Corpus Committee, New Investigator Committee, NCI Uterine Taskforce, national PI for NRG-GY020 and other trials under development, and in additional local roles as the Disease Specific Research Group Leader, NRG Oncology site PI for Gynecologic Oncology and Fellowship Director.

NIH Research Projects · FY 2024 · 2024-09

ABSTRACT Half of the spinal cord injury (SCI) individuals experience weight gain attributed to increased adiposity. Following SCI, increased lipid deposition is also observed in other organs including the liver, heart and skeletal muscle. Accumulating evidence indicates that pathological expansion of adipose tissue and ectopic lipid accumulation increases the risk of cardiometabolic disorders including hypertension, dyslipidemia and insulin resistance. Under physiological conditions, an excess of energy is stored in adipose tissue in the form of triglycerides. During energy deprivation, the adipose tissue mobilizes triglycerides into fatty acids via lipolysis, and these are used as fuel for brown adipose tissue, heart, liver, and muscle. When energy balance becomes dysregulated, ectopic accumulation of lipids in the liver and muscle as well as unresolved inflammation in adipose tissue contribute to insulin resistance. Whether the underlying cause of metabolic complications after SCI may be linked to adipose tissue dysfunction is unclear. Also unknown are the mechanisms that cause or contribute to pathophysiological changes in white adipose tissue (WAT) structure and function after SCI. The sympathetic nervous system and sensory neurons with cell bodies located in dorsal root ganglia (DRG) innervate WAT. Whereas the contribution of sympathetic innervation to WAT function has been characterized, the extent to which sensory innervation drives WAT function under normal and pathophysiological conditions has remained elusive. We and others have demonstrated a remarkable convergence between neuronal circuits' structural and functional organization and expression of α2δ subunits of voltage-gated calcium channels (VGCC). α2δ subunits positively regulate synaptic transmission by increasing plasma membrane expression of VGCC. However, these subunits may also play a pathological role following neuronal injury. Our preliminary data suggest that SCI exacerbates lipolysis in epididymal (e)WAT and that α2δ1 expression increases after a thoracic SCI in calcitonin gene-related peptide (CGRP) DRG neurons that project to eWAT. Concurrently, we found elevated CGRP content in eWAT and increased expression of the CGRP receptor RAMP1 in eWAT seven days after SCI. Not only does CGRP activate lipolysis independent of sympathetic drive, but it also acts as a potent vasodilator. Changes in adipose tissue microcirculation have been associated with health complications including adiposity and altered metabolism. Accordingly, experiments in Aim 1 will evaluate the extent to which α2δ1 silencing and pharmacological blockade normalize eWAT function after SCI. Aim 2 will dissect the cellular mechanism underlying α2δ1-dependent changes in eWAT lipolysis. Aim 3 will explore the role of vascular cells in eWAT dysfunction and inflammation after SCI. Successful completion of the proposed study may provide novel insight into molecular causes and mechanistic underpinning of maladaptive sensory processing and WAT function after SCI, facilitating the development of strategies targeting WAT function to reduce metabolic and cardiovascular complications after SCI.

NIH Research Projects · FY 2025 · 2024-09

Childhood social anxiety disorder is a highly prevalent and impairing condition; in severe cases it can lead to school refusal and social isolation and is often comorbid with other mental and physical health problems posing a significant public health burden. Although exposure to environmental toxicants have not commonly been thought of as contributors to mental health problems, epidemiologic studies have linked prenatal exposure to ambient air pollution with increased risk for anxiety symptoms and disorders. Children living in the context of economic disadvantage are at disproportionately higher risk for anxiety and exposure to air pollution. This proposal integrates findings from environmental health sciences and developmental psychology/neuroscience and proposes a novel framework detailing the pathway through which prenatal exposure to air pollution contributes to SADS in those children most at risk. In animal models, prenatal exposure to polycyclic aromatic hydrocarbons (PAH), a common and neurotoxic class of air pollutants, causes increased avoidance behaviors, analogous to human behavioral inhibition (BI), a well-documented risk factor for SADS. Prenatal PAH exposure also alters children’s cognitive control, which in turn increases SADS risk in children with BI. We have linked prenatal exposure to PAH with BI-like behaviors in infants and children and with internalizing symptoms and altered cognitive control. We now propose to test the overarching hypothesis that (1) prenatal PAH exposure is linked with SADS via effects on BI-like behaviors and (2) that prenatal PAH exposure alters neural function underlying cognitive control, and (3) moderates the association between PAH, BI, and SADS. We will conduct the study in 200 children living in the context of economic disadvantage who are followed in a prospective longitudinal birth cohort at the Columbia Center for Children’s Environmental Health. Using EEG to measure alterations in brain function associated with PAH exposure will allow us to show effects of air pollution on children's task-related brain function (versus structure) for the first time and at very early time points (preschool age) when intervention and prevention may be most effective. Findings will identify modifiable inflection points in developmental cascades that can be targeted to deliver maximally effective personalized intervention strategies. In addition, working collaboratively with implementation scientists and public health agencies, our findings can be used to develop personalized prevention strategies targeted to help children who are at risk for exposure and poor mental health outcomes.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract This project aims to determine whether a media-based intervention targeting low- socioeconomic status (SES) preschoolers and their caregivers can improve children's early literacy skills relative to a caregiver-led shared-reading intervention, in line with two NICHD priority areas: (1) effects of technology and media on child development and (2) school readiness in children from low-SES homes. The current proposal is situated in an implementation science framework, aiming to identify barriers to implementing evidence-based practices and adopting strategies to address these barriers and improve outcomes. In prior work, we found that a storybook intervention with explicit print referencing improves children's early literacy skills; however, caregiver adherence is often low. Here, we hypothesize that using educational media may circumvent or diminish barriers and promote adherence, as media may be more readily incorporated into household routines and thus increase child exposure to print references, a key mechanism for learning. This study addresses four specific aims: (1) To determine the extent to which a media-based, caregiver-led early literacy intervention leads to higher adherence among low- SES families relative to an aligned shared reading intervention (and the extent to which the media intervention reduces barriers, which then leads to higher adherence), (2) To determine the extent to which a media-based, caregiver-led early literacy intervention leads to greater improvement in literacy skills of children from low-SES homes, relative to an aligned shared reading intervention, (3) To determine the extent to which effects are mediated by higher adherence (i.e., intervention dose and intensity), and (4) To determine the extent to which two potential moderators (child sex and caregiver self-efficacy beliefs) influence effects of intervention medium. We also explore potential child-, caregiver-, and dyad-level moderators of intervention medium on early literacy skills. This study uses a randomized controlled trial with three conditions: a caregiver-led media-based early literacy intervention, a caregiver-led shared reading early literacy intervention, and a business-usual control. Participants are 450 caregivers and 3.5 to 4-year-olds. Early literacy skills are assessed at pretest and posttest. Social validity of the intervention is measured to test whether the media intervention circumvents barriers. Fidelity of implementation is measured during the intervention. Follow-up visits at three, six, and twelve months tests long-term effects. Aim 1 uses regression models to compare adherence and social validity across conditions, and to predict adherence from social validity. Aims 2, 3, and 4 are addressed via regression models comparing gains from pre- to post-test and two-level growth models examining growth in skills across follow-up time points. This project is innovative as it seeks to understand mechanisms by which using media to address barriers to a caregiver-led literacy intervention may be effective, capitalizing on the benefits of both educational media and caregiver-led interventions and testing mechanisms underlying effects on literacy skills.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Cellular metabolism instructs immune cell functions in both physiological and disease conditions. Activation of immune cells in response to invading pathogens induces a robust metabolic reprogramming, which has been indicated to play a critical role in orchestrating immune cell functions. The possibility of modulating immune cell function through metabolic regulation represents a new strategy to strengthen host defense mechanism. However, the role of individual metabolic pathways and enzymes during pathogen infection is not well understood, which represents a critical knowledge gap. Recently, we have identified the O-GlcNAc transferase (OGT), an important glucose metabolism enzyme, as a critical regulator of innate immune signaling during virus infection and septic inflammation. OGT mediates a ubiquitous post-translational modification (PTM) of cytosolic and nuclear proteins, known as O-GlcNAcylation. OGT-mediated O-GlcNAc signaling is well known as an essential regulator of many fundamental aspects of cell physiology and dysregulated O-GlcNAc signaling occurs in numerous human diseases. Despite a broad pathophysiological significance of OGT, the function of OGT in host defense mechanism against pathogen infection is just emerging and still underappreciated. Our preliminary results indicate that OGT directly restricts intracellular replication of both virus and bacteria, in addition to its previously established impacts on regulating innate immune signaling. By studying cell and mouse models with either OGT gene-deletion (Ogt-KO) or enzyme-inactive OGT (OgtK908A), we observed both enzyme-dependent and -independent functions of OGT in modulating innate immune signaling and limiting intracellular pathogen replication. We hypothesize that OGT serves as a central component of host defense mechanism by sensing pathogen infection and integrating innate immune signaling and cellular lipid metabolism via both enzyme activity-dependent and -independent manner. The long-term objective of our work is to determine the principles, functions and mechanisms of OGT and O-GlcNAc signaling in health and diseases. In this proposal, we aim to use our newly developed OGT enzyme-inactive mouse model and OGT gene-deletion model to understand the molecular mechanism by which OGT mediates host defense responses against invading virus and bacteria. We will advance this goal through three directions: 1) how does OGT sense virus infection and integrate innate immune response and cellular fatty acid synthesis pathway? 2) what is the functional importance of OGT-mediated protein O- GlcNAcylation in the detection of intracellular bacteria? and 3) how does the ubiquitination of OGT affect host defense? This study will significantly advance the field of immunometabolism by elucidating the function and mechanism of an important metabolic enzyme sensing and antagonizing pathogen infection. Successful development of OGT enzyme-inactive mouse model will also benefit other research topics such as cancer, diabetes and neurodegenerative diseases.

NIH Research Projects · FY 2025 · 2024-09

PROJECT ABSTRACT Rural Appalachian counties have the highest cancer incidence and mortality rates in the United States (US). While most cancer disparities are narrowing between Appalachian and non-Appalachian areas of the US, disparities in lung and oral cancers are widening. A high prevalence of cigarette smoking (i.e., >30% in many rural Appalachian counties vs. 11.5% nationally) primarily contributes to these disparities. As cultural factors, deficiencies in the rural Appalachian healthcare system, and tobacco industry actions have undermined the success of tobacco control approaches that proved successful elsewhere, we propose a new approach to reduce the prevalence of cigarette smoking in rural Appalachia. Oral nicotine pouches (ONPs, e.g., Zyn, On!), which contain nicotine but no tobacco, have a similar toxicant profile to medicinal nicotine replacement therapy (NRT), with most carcinogens and toxicants below the limit of detection. ONPs deliver nicotine similarly to NRT lozenges but are about half as expensive, are marketed to smokers, and our preliminary data indicate that rural Appalachian smokers find them more socially acceptable than cigarettes and more palatable than NRT. We propose a 6-month, two-arm, remote, randomized switching trial of 1,000 adult smokers who live in rural Appalachia. Participants will be randomized to either an ONP or an NRT (patch and lozenge) arm. During a 2- week Sampling Phase, participants will sample products in their relevant study arm in a range of flavors and nicotine concentrations and will select the flavor (ONP arm) and nicotine concentration (ONP and NRT arms) for use during the Switch Phase. During a 12-week Switch Phase, participants will attempt to stop smoking cigarettes and completely switch to their study ONP or NRT using study products provided for free. During a 14-week Observation Phase, participants will no longer receive study products but will be followed to evaluate longer-term switching outcomes. Participants will self-report smoking and study product use on online surveys, and changes in cigarette smoking will be confirmed using remote carbon monoxide assessment. This significant and innovative study will be the first to evaluate whether ONPs are suitable substitutes for cigarette smoking among rural Appalachian adults. If our study finds that ONP participants have greater success in reducing or quitting smoking than NRT participants, results will provide a foundation for interventions and regulations that support switching from cigarette smoking to ONP use. If our study finds that ONPs provide the same (or worse) effects on smoking outcomes as NRT, results will suggest that public health practitioners, regulators, and clinicians should steer smokers away from ONPs in favor of NRT.

NIH Research Projects · FY 2025 · 2024-09

Project Summary The Clinical Trial Concept in the Oncology Landscape (CTC) is a virtual and in-person, hybrid course designed to educate oncology trainees and junior faculty from any cancer specialty in applying Good Clinical Practice (GCP) and scientific reasoning into effective clinical trial design. The course will accept junior oncologists and fellows/residents in an oncology specialty, who are novice in cancer clinical research. The course will deliver in Part 1, The Fundamentals of Clinical Research, a semester long virtual immersive educational experience across all areas of cancer clinical research, then culminate in an in-person Trial Concept Writing Camp (Part 2) during the annual American Association of Cancer Institute (AACI) Clinical Research Innovation (CRI) meeting where participants will be mentored by experienced clinical investigators from different AACI member institutions during which time, they will finalize a clinical trial concept (also called letter of intent (LOI)), the end product of this course. Participants are selected through a competitive application process consisting of a CV, personal statement, letter of support from a local clinical research mentor, and a career development plan. Accepted participants will first receive scientific and GCP training over 20 virtual sessions covering the 6 major modules of protocol development that encompass the eleven domains of clinical trial expertise, including oncology, science, medicine, business/negotiation, accounting, statistics, pharmacy, pharmacology, laws and regulations, ethics, and clinical trial management. The curriculum will be loaded on a web portal developed for educational purposes under another U24 NCI award to OSUCCC. The virtual training will use a flipped classroom approach to maintain participant attentiveness. At the CRI meeting, participants will interact with Faculty mentors, nationwide experts in clinical research, who will help them complete a realistic concept form that meets scientific standards. Participants will take part in pre- and post-tests to determine the cogency of the CTC curriculum for their professional practice, and the effectiveness of the course logistics. They will complete the course by submitting a finalized clinical trial concept to the CTC course directors. Afterwards, they should be able to use their concept to develop a full protocol at their own institutions or though the NCI clinical research networks or other clinical trial networks. After the course, public data will be used to track participants’ career trajectories to determine the long-term effects of the course on their careers, working with Academics Analytics, a company contracted with OSU. Impact. Investigators must apply GCP and the International Conference on Harmonisation (ICH) guidelines to the design and scientific approach of their clinical research. The basics of clinical research are not taught in medical schools or residencies. The pool of clinical investigators has also decreased in the US, creating a serious shortage of clinical trial experts. At the same time, there has been an explosion of biological discoveries and technological advances. This innovative and necessary CTC course will address the need for thorough and specific training on clinical trial design and execution to eventually benefit patients with cancers and support the professional development of junior oncology faculty.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Oral cancer is a significant public health problem, as 58,450 new cases and 12,230 deaths occur annually. in the United States. Despite treatment advances, the overall survival rates (60% at 5 years, 50% at 10 years) for oral cancers have not appreciably changed over the past 30 years. Therefore, new and effective oral cancer chemoprevention and treatment strategies are needed. Although synthetic glucocorticoids are routinely used as adjuvant therapy in postoperative oral cancer patients, recent evidence indicates that glucocorticoids may actually promote tumor progression in some cases, thereby challenging the use of these compounds in patients with premalignant and malignant oral disease. Since glucocorticoid biosynthesis and regulation can occur in the oral mucosa, it is essential that we clearly understand the role of the oral glucocorticoid system on oral cancer development and metastasis. Further, we recently identified a potent cancer chemopreventive agent, gallic acid, which modulates glucocorticoid metabolism, but specific mechanisms associated with gallic acid mediated regulation of glucocorticoid metabolism during oral cancer chemoprevention is currently unknown. In this proposal, we propose to determine the mechanistic role of the oral glucocorticoid system on oral cancer development and metastasis and determine the effects of the chemopreventive agent gallic acid on the glucocorticoid metabolic pathway during oral cancer chemoprevention. We will use oral cancer cell lines, animal models and oral cancer clinical samples to determine these effects. Our hypothesis is that downregulation of the glucocorticoid inactivating enzyme, Hsd11b2, promotes active glucocorticoid signaling in the oral mucosa, which promotes cancer, and inhibition of this pathway by gallic acid can prevent oral cancer development and metastasis. Studies in aim 1 will define the impact of glucocorticoids and Hsd11b2 on oral cancer development and metastasis. We will use genetically modified mice, cell lines and patient samples, to determine how glucocorticoid metabolism affects oral carcinogenesis. Studies in aim 2 will determine the effect of glucocorticoid signaling on tumor cells and tumor infiltrating T cells during oral carcinogenesis. We will use genetically modified mice targeting the glucocorticoid receptor Nr3c1 on oral cancer cells and T cells for these studies. Studies in aim 3 will determine the effect of gallic acid on oral glucocorticoid metabolism during oral cancer, and how the regulation of glucocorticoid metabolism by gallic acid affects oral cancer outcomes. The complementary expertise of our investigative team coupled with novel experimental approaches will facilitate the successful completion of the aims proposed in this application. These studies will increase our understanding of how the oral glucocorticoid system affects oral carcinogenesis and the mechanisms of gallic acid mediated inhibition of oral cancer. Our studies also have the potential to change the current clinical practice of using synthetic glucocorticoids in oral cancer management.

NIH Research Projects · FY 2025 · 2024-09

SUMMARY Despite significant advances in Immuno-Oncology (IO) treatments, lung cancers with loss-of-function mutations in STK11, comprising 20-30% of cases, demonstrate poor prognosis due to resistance to these strategies. Our proposal targets the interaction between specific tumor 'differentiation phenotypes' and immune resistance pathways, associated with STK11-Deficient lung cancers. Firstly, we've found that these differentiation phenotypes drastically influence the immune composition of the tumor microenvironment (TME) and the patient's response to immune checkpoint blockade (ICB). Furthermore, we've developed STK11-MLDD, a novel biomarker that measures these differentiation phenotypes and may predict ICB benefit. Secondly, we've identified the immune regulator TRAF2 and its partner cIAP1 as promoters of immune resistance, specifically in the context of STK11 deficiency, and appear to be linked to the neuroendocrine differentiation phenotype. These findings provide the foundation for our two complementary aims. Aim 1 focuses on characterizing the interaction between tumor differentiation phenotypes, corresponding immune TME phenotypes, and clinical response to ICB. We'll validate the STK11-MLDD biomarker in multiple patient cohorts and develop it as a clinical tool for future trials. Additionally, using flow cytometry and single cell RNAseq data, we will identify specific suppressive immune cell populations and resistance phenotypes in the TME associated with different STK11-MLDD differentiation subsets. Aim 2 centers on understanding TRAF2/cIAP1-mediated immune evasion and validating TRAF2 as a potential clinical target in lung cancer. We'll conduct experiments evaluating changes in TNF signaling resulting from STK11 loss and TRAF2/cIAP1, assessing their impact on anti-tumor immunity. Additionally, we will conduct in vivo experiments to examine STK11-/TRAF2-mediated effects on the TME and the potential for TRAF2 inhibition to enhance ICB response. We anticipate that our results will establish TRAF2/cIAP1 inhibition as a promising clinical target for STK11- deficient lung cancer, while also defining novel mechanisms of immune resistance and additional precision treatment candidates for these cancers. Furthermore, the STK11-MLDD biomarker is expected to have use not only as a clinical predictor of ICB response, but also potentially as a ‘treatment-selective’ biomarker that could be used to identify patients likely to respond to precision approaches such as cIAP1 inhibition. As a physician-scientist in thoracic oncology, my career goal is to become a successful independent academic investigator, focusing on STK11-mutated NSCLC to improve treatments and outcomes for patients with these cancers. We have outlined a detailed mentorship and career development plan that will complement my scientific approaches and promote mastery of skills necessary for my long-term success, with specific focus in the areas of 1) bioinformatics and biostatistics, 2) tumor immunology and immune oncology, 3) academic leadership. Achieving these goals will accelerate my success and maximize my long-term impact in the field.

NSF Awards · FY 2024 · 2024-09

Scientists detect exoplanets by using spectrographs that measure tiny changes in the color of starlight caused by stars wobbling back and forth as planets orbit around them. In 1995 this method was used to detect the first exoplanet in orbit around a star like the Sun, a breakthrough recognized with the Nobel Prize in Physics in 2019. Finding even smaller exoplanets in the future, including those like Earth, requires careful calibration of the spectrographs that make these measurements. With funds from NSF’s Major Research Instrumentation program, this team will procure, install, and optimize a new “laser frequency comb” calibration source for two spectrographs at the 8.4-meter Large Binocular Telescope in Arizona. In addition, this project will contribute to training the next generation of scientists and engineers, and graduate students will lead key program elements while being mentored by senior members of the team in training and career development. High resolution spectroscopic measurements will play an essential role in exoplanet science. In addition to supporting transit surveys, the radial velocity (RV) method will also play a critical role in the characterization of planets discovered by future direct imaging missions. To accomplish these goals it is crtical that we continue to advance the sensitivity, resolution, and precision of extreme precision radial velocity (EPRV) spectrographs. This project will deliver a state-of-the-art Laser Frequency Comb (LFC) to the Large Binocular Telescope (LBT) and complete its optimized integration, enabling critical multi-year spectroscopic studies with the Potsdam Echelle Polarimetric and Spectroscopic Instrument (PEPSI) and iLocater spectrograph. With their precision high-resolution (R>190,000) capabilities spanning visible (PEPSI: 383-912nm) and near-infrared (iLocater: 970-1310nm) wavelengths on a dual 8.4m telescope, these instruments are optimized for EPRV studies of exoplanets and stellar astrophysics. Augmenting them with a dedicated LFC with its absolute wavelength calibration capabilities enables robust intercalibration between LBT instruments and expands short-term instrument performance to multi-year baselines. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

Stellar spectra are the primary way we learn about stars, including their composition, age, and special features like star spots or hidden companion stars. Although we can now observe millions of star spectra, our current methods aren't robust enough to fully explain what this starlight reveals about stars. This project aims to resolve this problem: how to connect the latest computer models of stars with the vast number of stellar spectra we've collected. The researchers will use new artificial intelligence (AI) methods to create AI models. They will then combine these models with detailed computer simulations to analyze star spectra more effectively. This research aligns with national priorities, using AI to accelerate research and training new experts who are knowledgeable in both scientific research and AI. The investigators will develop spectral foundational models—robust AI models pre-trained on vast numbers of simplified stellar spectral models. These models will be designed to capture the underlying atomic and plasma physics of stellar atmospheres through carefully crafted training tasks. While simplified one-dimensional physical models assuming spherical symmetry are easy to generate, they lack nuance. Conversely, detailed three-dimensional models with full non-hydrostatics, though accurate, are resource-intensive and can only be generated sparingly. To address this challenge, the researchers will harness the latest Transformer-based neural network models, establishing methods for pre-training these models with robust unsupervised tasks aimed at teaching the models to grapple with the underlying complex physics in stellar atmospheres. Building upon the AI foundational model, the team will integrate it with vertex frameworks and probability density estimators with neural networks. The research team will examine two specific science cases. First, they will elucidate star-spot distributions using spectra from the Sloan Digital Sky Survey V (SDSS-V). Second, they will infer characteristics of stellar populations, including age, mass, and metallicity distributions, from galaxy spectra collected by the Dark Energy Spectroscopic Instrument (DESI). This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

The ability to understand other people’s thoughts and feelings (commonly referred to as theory of mind) is related to narrative comprehension during the preschool years because children need to understand characters’ mental states to understand the meaning of a story. Little research has explored what predicts how these two skills develop together. This project examines two family-level factors, the home literacy environment and the mental state talk parents use when reading to children, that might explain how theory of mind and narrative comprehension co-develop during early childhood. Both skills are important for school readiness because theory of mind predicts socially competent interactions with peers, and narrative comprehension predicts later reading comprehension. Thus, understanding the predictors of theory of mind and narrative comprehension, and how they are reinforced by the same early environmental supports (such as parent-child talk about mental states), is key to developing approaches to promote these skills. The research plan for this project includes testing preschoolers at three time points, each six months apart, using longitudinal three-way cross-lag panel design. At the first time point, the research team gathers information on the home literacy environment (such as how frequently parents read to children and the mental state language parents use when reading to children). At each time point, the research team tests preschoolers’ theory of mind and narrative comprehension skills. The study includes additional assessments to control for the possibility that other variables (including socioeconomic status and children’s vocabulary size) contribute to the observed relations between the key variables of interest. The research plan includes recruitment of families that are diverse with respect to race, ethnicity, and socioeconomic status. The study design allows the research team to test a novel framework that highlights shared developmental antecedents of theory of mind and narrative comprehension skills. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

When machine learning (ML) and artificial intelligence (AI) techniques are used in applications involving humans (e.g., recommending personalized items to users, ranking candidates for admission, hiring, and lending), it is critical to ensure safety for both the learning system and humans. From the learner’s perspective, the ML system should prevent manipulated information from disrupting the training procedure (safe training) and remain robust against rare and unexpected events during deployment (safe deployment). From the human standpoint, it is crucial that ML decisions align with social values (safety perception) and prevent the system from evolving toward unsafe states (safe downstream effects). However, achieving such safety assurance is often challenging due to the complex interactions and feedback dynamics between humans and the learning system. For instance, humans who utilize for obtaining loans or job searches may change their behavior, such as changing their profiles, to achieve favorable outcomes. While digital platforms offering on-demand services may steer consumer preferences to benefit their service. Meanwhile, as the users evolves, the learning system needs to update accordingly. Under such intricate human-AI interactions, creating a safe learning environment that supports long-term human well-being remains a significant challenge. This project aims to develop theoretical and algorithmic foundations for building a human-AI ecosystem with long-term safety assurance. The outcomes have the potential to benefit diverse domains, including lending, recruitment, healthcare, admission, and recommendation systems. To achieve long-term safety in the human-AI ecosystem, the project explicitly considers the complex interactions between humans and the learning system, with a research agenda comprising the following objectives: 1) Develop an analytical framework to characterize human-AI interactions, which embeds all safety components for both the learner and human agents; 2) Examine the feedback effects between agents and the ML system, developing methods to ensure the long-term safety of both under their dynamic interactions; 3) Establish a causal understanding of human-AI dynamics and design transparent and interpretable interventions to achieve long-term safety. This agenda entails developing new theories and algorithms at the intersection of control theory, reinforcement learning, dynamical systems, and optimization. Beyond theoretical and algorithmic contributions, the project will be validated through various use cases, including recommendation systems, lending, and healthcare. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

By the end of this century, sea level is expected to rise by one meter due to climate change and inundate coastal areas that are home to millions of people in the United States. As sea levels rise, groundwater levels will also rise, and groundwater salinities will increase, causing damage to subsurface infrastructures such as tunnels, building foundations, and stormwater and sewer systems. Until now, coastal risk research has largely excluded groundwater-related hazards because they are harder to observe than floods that occur above ground. The long-term vision for this project is to advance new methods for mapping risks to subsurface infrastructure in coastal communities due to sea level rise. The project will co-develop new approaches for assessing exposure and vulnerability to shallow groundwater hazards so that communities can use maps to understand potential damage from rising and salinizing groundwater and make informed decisions about how to maintain or surrender subsurface infrastructure. One important outcome of this planning proposal will be the formation of a Boston area coalition of scientists and community leaders who are concerned about risks to subsurface infrastructure from sea level rise that will collaborate on new risk assessment methods. The coalition will be engaged in conversations about adaptation pathways for mitigating the effects of sea level rise. The activities and products from this planning project will catalyze a larger-scale proposal to extend risk assessment methods to new infrastructure categories and coastal communities throughout the United States. A postdoctoral scientist and PhD student will be mentored in a highly interdisciplinary endeavor that will generate new knowledge at the interface of geoscience, engineering, economics, and urban planning. This pilot-scale effort will focus on a specific subsurface asset category, such as stormwater and sewer systems, and apply workflows to at least two distinct testbed communities in the Boston area, which encompasses the densely urbanized shoreline of Boston Harbor and a diverse set of ex-urban communities. One anticipated product will be pilot hazard, exposure, vulnerability, and impact (or risk) maps. The project will also generate a clearer picture of challenges to upscaling or transferring the workflow to other asset categories and communities. Lastly, a workshop will be convened where prototype risk maps will be used to engage community leaders in adaptation planning conversations to mitigate the impacts of sea level rise on coastal communities. These experiences will inform a future proposal to develop risk maps for new geographic areas and engage communities in adaptation planning. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

The modern oceans and the ecosystems they contain resulted from millions of years of change in physical and biological ocean systems. One aspect of the environment that has a large impact on marine animals is the amount of available food and nutrients. Understanding how individual organisms and biological communities adapt and respond to changes in nutrient availability advances scientific knowledge by 1) improving understanding of how the physical environment drives evolution, and 2) providing insight into how decreased nutrients might trigger regional extinction events. These results are important for understanding the geologic history of life as well as its future. In addition to these scientific objectives, this project supports the training and advancement of students through 1) an inclusive field course for advanced undergraduate students, 2) the development of a graduate student cohort trained to participate in international field research, and 3) the production of a bilingual graphic novel to increase scientific literacy in K-12 students in the US and the Caribbean. The goal of this project is to understand and characterize the relationship between surface productivity and ecological structure in marine benthos by (1) evaluating how productivity affects the energetic and trophic structure of marine benthic communities on both sides of the modern Isthmus of Panama, (2) using this knowledge to evaluate the fossil record of Caribbean benthic ecosystems before, during, and after the uplift of the isthmus, and (3) relating ecosystem changes driven by productivity shifts to the well-documented Caribbean extinction event ~2 Ma. The project leverages collections from the Panama Paleontology Project, which includes extensive collections of modern mollusks and rich fossil collections, to meet these objectives. The project applies new technologies, including high-throughput imaging and automated morphometric methods, to analyze the size-frequency distribution and calculate measures of energetics. The project also explores how trophic composition, larval dispersal mode, and predatory attack frequencies in mollusk shells in modern death assemblages and fossil assemblages vary across productivity gradients. The project will advance the community’s current understanding of how these ecological traits are influenced by productivity in modern systems and the role they have played in the evolution of modern Caribbean ecosystems. The Biological Oceanography Program co-reviewed and co-funded this project with the Sedimentary Geology and Paleobiology Program. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-09

Project Summary: Our organs respond dynamically to changes in the body’s demands by adapting both their size and activity levels. In these processes, mechanisms governing cell fate decisions, which influence the equilibrium between cell death and cell proliferation, assume crucial roles. Although decades of research have revealed the involvement of cell cycle regulators, epigenetic modulators, metabolic pathways and ionic homeostasis in this complex decision process, we still have major gaps in our understanding of the mechanisms that govern cell fate. These gaps need to be addressed in order to better understand the pathophysiology of many life threatening conditions stemming from the errors of cell fate decision, such as uncontrolled cell death seen in degenerative diseases or uncontrolled cell proliferation in neoplasms and autoimmunity. Our recent work showed that B cell fate is regulated through two spatiotemporally distinct signals, 1- by antigen binding to the B cell receptor (BCR), and 2- via T cell help or pathogen-driven activation of the toll-like receptors (TLRs). While signal-1 primes the cell for activation and proliferation, it also initiates a short countdown to death that can be halted by a “signal-2”. The presence of signal-2 promotes cell survival and proliferation by validating the accuracy of the signal-1, while its absence causes a gradual increase in intracellular calcium leading to mitochondrial dysfunction and cell death. We called this process the ‘metabolic clock’. Because mitochondria and calcium homeostasis have been shown to play roles in determining the fate outcomes of many different cell types; our metabolic clock model provides an excellent platform to dissect the mechanisms that govern life and death decisions throughout the body. The overarching goal of our proposal is to unveil the interplay between signals -1 and -2 at the molecular level, with a view towards identifying how metabolic clocks regulate cell fates. Our first project will define how signal-1 induces calcium increase, the main driver of mitochondrial dysfunction, and how this can be prevented by signal- 2 to promote survival and proliferation. Our second project will delineate whether mitochondrial remodeling, under the influence of signals -1 and -2, can modulate the metabolic clock by altering the sensitivity of mitochondria to rising cytoplasmic calcium. Lastly, we will reveal whether the metabolic clock may lead to cell fates other than death or proliferation, such as the induction of an hypofunctional anergic state. We will use transgenic mouse models, novel bone marrow chimeras and human samples to perform integrated sets of biochemical and cellular assays, unraveling how the two signals independently or in combination impact activation of signaling molecules, mitochondrial remodeling, calcium dynamics, and energy production pathways. Altogether, this proposal, bridging basic mechanistic research with in vivo models, will provide a novel perspective of cell fate decision mechanisms, paving the way for new therapeutic approaches to disorders caused by the errors in this process.

NSF Awards · FY 2024 · 2024-09

The Generation-4 NSF Engineering Research Center titled “Transformation of American Rubber through Domestic Innovation for Supply Security: TARDISS” will lead fundamental research towards US natural rubber biomanufacturing. Currently the single commercial source of natural rubber is the tropical rubber tree (Hevea brasiliensis), with production areas all outside of the United States. TARDISS will use a systems engineering approach to integrate engineering with biology, biotechnology, agriculture, and other disciplines optimizing alternative plants to produce entirely new natural rubber materials at scale. The TARDISS team will collaborate with communities, farmers, processors and rubber manufacturers to enable biomanufacturing-based natural rubber production optimized to large parts of the US, with a focus on marginal agricultural lands. TARDISS will enable a circular biomanufacturing economy that respects natural systems, including pollinator services by the new domestic crops, water recycling and re-use, additional CO2 capture, and an estimated 2 million jobs tied to US soil. Engineering workforce development will provide training in this new U.S. area, also include those with untapped potential that are currently underutilized in the workforce. The outcomes will be a sustainable domestic rubber industry and a new, young workforce converging engineering and agriculture trained through a new American Rubber Academy. The “U.S. Sunlight to Materials” vision motivating the systems engineering approach of TARDISS is encapsulated by two hypotheses: #1: The U.S. can replace imported natural rubber (NR) with rubber from domestic crops, utilizing marginal agricultural lands, hydroponic systems, and new extraction methods; and #2: The U.S. can replace imported goods with products made with home-grown natural rubber. TARDISS will integrate engineering with biology and other science disciplines via the following three research thrusts: 1: BioEngineering will converge engineering, biochemistry, enzyme chemistry, and molecular biology to fundamentally understand how plants naturally produce rubber. Natural variety will be combined with genetic approaches to tailor hydroponic dandelion to produce new NR variants and transfer the knowledge to the guayule and mountain gum plant species. 2: Crop Engineering will converge plant and agricultural engineering to develop and disseminate new “smart” crop production practices for all three crops. 3: Latex/Rubber Engineering will converge engineering, materials/polymer science and engineering, chemistry, and physics to invent extraction methods to produce consistent high-performance latex and rubber and new processing methods for products. Furthermore, TARDISS will invent enabling technologies in field and hydroponic systems, industrial scale latex and rubber extraction methods, and novel processes-for-products and bring these to communities. A seamless integration of scalable biology, engineering, and science, while co-developing economically scalable pathways with domestic stakeholders, will be critical for success in Convergent Research. The outcomes will be a sustainable domestic rubber industry and a new, young workforce converging agriculture and engineering trained through a new American Rubber Academy in collaboration with the Rubber Division of the American Chemical Society. The TARDISS Innovation Ecosystem will bring together American business leaders and entrepreneurs, researchers, students, national labs, and communities, and features novel programs such as the Piranha Pit to encourage innovation. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.