Ohio State University

NIH Research Projects · FY 2026 · 2024-02

Mutations exclusively in SLC29A3, which encodes the lysosomal transporter termed equilibrative nucleoside transporter-3 (ENT3), cause an expanding spectrum of human genetic disorders, such as H syndrome, PHID syndrome, RDD syndrome, SHML syndrome, dysosteosclerosis etc. They share common mutations and overlapping clinical manifestations with anemia, erythroid hypoplasia and hepatosplenomegaly as characteristic signs. Our group has recently described an indispensable role of ENT3 in the maintenance of hematopoietic stem cell (HSC) homeostasis via the genetic deletion of ENT3 in mice. Intriguingly, ENT3- deleted mice manifest clinical signs closely resembling human SLC29A3 disorders. The objective of this application is to evaluate the molecular disease pathogeneses and treatments of SLC29A3 genetic disorders. Specifically, it is our hypothesis that interference with the lysosome-to-ER mobilization of bile acid (BA) chemical chaperones and ER stress signaling in HSCs underlies the pathogeneses of SLC29A3 dysfunctions and that endogenous or synthetic chaperones will serve as suitable treatment agents to overcome these disorders. This hypothesis is based on preliminary data showing that BA chemical chaperones are novel cargos of ENT3, showing ENT3 conferring the BA-dependent amelioration of ER stress signaling in HSCs, and showing improved function of ENT3-deleted HSCs after treatment with salubrinal, a chemical ER stress reducer. The rationale for this project is that the identification of the mechanisms of ER stress regulation by ENT3 in HSCs will determine the molecular disease pathogenesis of SLC29A3 disorders, which would provide therapeutic opportunities to treat the SLC29A3 genetic disorders. This central hypothesis will be tested by pursuing two specific aims. Specific Aim 1 will evaluate ENT3 transport and the subcellular disposition of BA chemical chaperones in HSCs. The working hypothesis is that ENT3 promotes the lysosome-to-ER mobilization of BA and that the loss of ENT3 will reduce the ER accumulation of BAs with increased sequestration in the lysosome. We will use new investigational models, ENT3 structure-function analysis, novel BA molecular probes, time lapse imaging, subcellular pharmacokinetics, and mass spectrometry studies to address this aim. Specific Aim 2 will evaluate aberrant ER stress signaling as the basis of HSC dysfunction in SLC29A3 disorders. Key questions connecting ENT3-regulated ER stress signaling and SLC29A3 disease pathologies will be evaluated in steady-state and stressed HSCs using HSC-specific, inducible and conditional ENT3 KO mice. Furthermore, endogenous and synthetic chaperones and pharmacological modifiers of ENT3 misfolding, trafficking, and degradation will be evaluated to identify suitable treatment agent(s) for overcoming SLC29A3 disorders. Completion of this project will illuminate the pathogenetic basis and treatments for SLC29A3 genetic disorders and will provide unique insight into the ENT3-regulated BA chaperone defense mechanisms in the physiological ER stress signaling in HSCs.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY (ABSTRACT) Ozone (O3) is a harmful air pollutant that exacerbates chronic lung diseases in part by activating inflammatory responses, inducing lung injury, and inhibiting resolution mechanisms. Resolution mechanisms are mediated largely by specialized pro-resolving mediators (SPMs). SPMs are potent bioactive molecules that inhibit immune cell recruitment, downregulate pro-inflammatory cytokine and chemokine production, and upregulate phagocytosis of apoptotic cells – a resolution process termed ‘efferocytosis’. We have previously reported that O3 decreases SPM production and inhibits efferocytosis, potentially contributing to exacerbated and persistent inflammation observed in patients with pulmonary diseases exposed to O3. SPMs are metabolized primarily from ω-3 fatty acids, such as docosahexaenoic acid (DHA). DHA can be consumed through diet or, more commonly in western cultures, synthesized from the essential fatty acid α-linolenic acid by elongation of very long-chain fatty acids protein 2 (ELOVL2). Preliminary data in this proposal indicates that DHA concentrations and ELOVL2 expression increase in the lung tissue following O3 exposure. Furthermore, ELOVL2 is increased in alveolar macrophages (AMs) after O3 exposure, suggesting an immunological need for endogenous DHA in AMs. When exogenous DHA was supplemented through diet, the O3-induced pulmonary and AM driven inflammatory responses were reduced, AM efferocytosis was augmented, and markers of lung injury resolved faster. Additionally, similar findings were noted when mice were pretreated with DHA derived SPM intermediates that correlated with an increase in the SPM maresin 1 (MaR1) in lung tissue. MaR1 is a unique SPM that is primarily produced by monocytes and macrophages that binds to leucine rich repeat containing G protein-coupled receptor 6 (LGR6) to facilitate resolution processes. To examine MaR1 signaling following O3 exposure, we measured LGR6 expression on AMs and found it to be significantly reduced when pulmonary inflammation was ongoing. Taken together, these data indicate that endogenous DHA in AMs leading to MaR1 signaling may be a crucial pathway to resolving O3-induced lung injury and inflammation. Therefore, we hypothesize that endogenous DHA synthesis in AMs is required for production of MaR1 which reduces the severity and improves resolution of O3-induced lung inflammation . To test this hypothesis, we will: 1) Examine how endogenous DHA synthesis via ELOVL2 in AMs reduces O3-induced pulmonary inflammation and promotes resolution responses, and 2) Determine if DHA-derived MaR1 signaling induces a pro-resolving AM phenotype through LGR6, mitigating pulmonary inflammation and promoting resolution following O3 exposure. Completion of the proposed aims and training included in this proposal will prepare the applicant for a successful career as an independent investigator and will equip him with cutting edge techniques, a strong network of mentors, and didactic training required for a successful career in academia.

NIH Research Projects · FY 2026 · 2024-02

Project Summary Obesity is a risk factor for pancreatic ductal adenocarcinoma (PDAC), a cancer that has a dismal 5-year survival rate due to limitations in prevention and treatment. Obesity promotes PDAC tumorigenesis and worsens survival by increasing systemic inflammation, metabolic dysfunction, and chemotherapy resistance, but the mechanisms that drive tumorigenesis in obesity are unknown. Deleting lipocalin 2 (LCN2) in obese genetically engineered mouse models (GEMMs) of PDAC decreases inflammation in the tumor microenvironment and increases survival. LCN2 can bind lipid ligands like fatty acids (FAs), which are dysregulated in the plasma and adipose tissue of obese PDAC subjects. Linoleic acid can suppress pro- inflammatory signals through several pathways, including inhibiting LCN2’s binding to matrix metalloproteinase-9. We observe that linoleic acid was lower in the plasma and arachidonic acid levels to be highly abundant in the adipose tissue of obese PDAC patients. Arachidonic acid can be derived from linoleic acid and then converted into pro-inflammatory prostaglandins by the enzyme cyclooxygenase 2 (COX2). We have previously shown that inhibiting COX2 in obese PDAC GEMMS reduces PDAC. Through other metabolic pathways, linoleic acid can be metabolized into anti-inflammatory prostaglandins. Therefore, understanding how LCN2, linoleic acid, and COX2 function in inflammation and metabolism in obesity may help improve prevention and treatment of PDAC. Our long-term goal is to determine how excess adiposity and inflammation due to obesity contributes to PDAC to identify targets for novel prevention and treatment strategies. HYPOTHESES: 1) Signaling from adipose and immune cells in obesity drives tumor development through changes in inflammation and FA metabolism in the adipose and tumor microenvironments. 2) Targeting FA metabolism will slow the development of obesity-associated PDAC and decrease adipose tissue inflammation. To test our hypothesis, we propose the following aims. AIM 1: Determine the adipose tissue and immune cell contributions of LCN2 signaling on obesity-associated PDAC. We hypothesize that increasing LCN2 signaling from excess adipose tissue and/or immune cells contributes to obesity-associated PDAC development, progression, and growth by modulating FA metabolism and inflammation in the tumor and adipose microenvironments. AIM 2: Determine whether targeting fatty acid metabolism inhibits obesity- associated PDAC. We hypothesize that increasing dietary linoleic acid levels and inhibiting COX2 will suppress tumors by modifying KRas activity and prostaglandin biosynthesis to promote anti-inflammatory signaling in PDAC. IMPACT: We will elucidate the signaling mechanisms that underlie how changes in the adipose tissue of obese individuals can drive the development and progression of obesity-associated PDAC. Understanding how adipose-derived factors contribute to obesity-associated tumorigenesis will lead to novel targets for improving PDAC prevention and treatment and provide a model for future work in other obesity- associated cancers.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY My career goal is to lead an independent research group to investigate the pathology and resolution of pain to solve neuropathic pain problems. To achieve this goal, I propose to develop and use novel translational strategies to investigate neuron-substrate interactions in the skin layer using a combination of human and animal models of peripheral sensory neuropathy. My research focuses on a common type of peripheral sensory neuropathy induced by chemotherapy, which affects over 3 million people in the US alone. While chemotherapy- induced peripheral neuropathy (CIPN) develops in up to 80% of cancer patients treated with chemotherapeutics, there are no effective prevention or treatment strategies. Considering the dynamic extracellular environment of cutaneous sensory nerves and recent understanding that keratinocytes can also function as a nociceptor, it is vital to understand the maintenance and function of both neurons and keratinocytes to solve neuropathic pain. Hence, I propose to test the hypothesis that dysregulation of neuron and epidermal interactions underlie pathological progression of CIPN. Building from my recent work that identified protective roles of integrins in CIPN, my proposal has three aims that use cutting-edge tools and innovative multi-disciplinary approaches. In Aim 1, I will investigate neuron-substrate interactions and their regulatory mechanisms in a Drosophila in vivo CIPN model. I will use genetics and advanced microscopy approaches to label, manipulate, and detect subcellular changes in neuron to substrate contacts and link with functional changes in a CIPN model. Using integrins as a model to understand cell surface protein-mediated mechanisms underlying CIPN, I will characterize endocytic regulators that modulate surface protein expression mediating neuron-substrate contacts. In Aim 2, I will use human induced pluripotent stem cell (iPSC) technology to identify keratinocyte-nociceptive neuron interactions and cell-type-specific pathology in a co-culture CIPN model. Given species differences in sensory neurons, I expect that establishing a human model to investigate the impact of sensory neurons and extrinsic factors on pathological progression in CIPN will provide critical insights. In Aim 3, I will establish and validate a 3D human skin-nerve co-culture CIPN model to investigate neuron-substrate relationships and integrin-mediated protection. Because subtypes of nociceptive neurons target different epidermal layers and contribute to distinct pathology in peripheral neuropathy, a 3D engineered human skin will be incorporated in the model. This will allow me to investigate subtype-specific pathology and to resolve nociceptive neuron terminal degeneration and dysfunction in CIPN. This proposal will provide new insights into pathological progression and mechanisms mediated by neuron-substrate interactions and a strong foundation for my future research. As my primary expertise is in neuroscience using Drosophila models, investigating human neurons and epidermal cells requires new training in skin biology and stem cell biology. The proposed plan will significantly facilitate my career goal to solve intractable neuropathic pain problems in patients.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY T2D is a major public health problem and is currently the 7th leading cause of death in the US. Despite a range of efficacious treatments, less than 50% of patients achieve a glycemic target of A1c < 7.0%, suggesting that this is due to difficulty with following medical regimens to reduce A1C levels. While a range of factors have been identified in this regard, we posit that a barrier to treatment are broad difficulty with emotional regulation that are not diagnosis-specific but lead to Diabetes Distress (DD) and difficulty in coping with medical regimens, and other aspects of diabetes self-care, in the context of the psychosocial stressors associated with T2D. Extant data suggests that sub-optimal emotional regulation (experience of intense emotion and skill at regulating emotion) is related to elevated DD and A1c levels, and that an Emotion-Focused Behavioral Intervention (EFBI) can reduce both DD and A1c levels in PWD with T2D. In this project we seek to, first, adapt our EFBI, initially developed as a one-to-one intervention, to a group intervention (G-EFBI) and, two, collect feasibility, acceptability, and preliminary efficacy data to determine if G-EFBI is a feasible, acceptable and, possibly, efficacious intervention compared to an “Attentional Control” intervention in PWD with T2D and elevated DD and A1c levels.

NIH Research Projects · FY 2025 · 2024-02

SUMMARY This proposal aims to generate a Cre driver in mice that can be used to investigate the distinct functions and contributions of type 3 innate lymphoid cells (ILC3s) in immune responses and diseases. ILC3s, similar to CD4+ helper T lymphocytes (Th), play crucial roles in host defense and in maintaining tissue homeostasis. However, the functional overlap between ILC3s and Th17 cells has made it challenging to study their specific contributions in these and other contexts. Our research team identified an ILC3-specific enhancer (E22-2) associated with the Il22 gene, which encodes a type 3 signature cytokine. We now propose to generate a mouse strain with ILC3- specific expression of GFP-IRES-Cre using CRISPR-mediated targeting of a single-copy BAC-Il22 reporter (22- GFP) containing E22-2 and exhibiting GFP expression that is exquisitely restricted to mature ILC3s. The specificity and extent of Cre-mediated deletion will be determined by crossing the ILC3-specific Cre mice (22- GFPCre) with a flox-STOP-TdTomato reporter strain. Successful completion of this project will facilitate genetic engineering and targeted manipulation to study the ILC3-specific functions of specific factors without confounding effects on Th17 or iNKT17 cells. Examples of such future studies include establishing the roles of type 3 cytokines (IL-22 and IL-17A/F) in controlling infections, inflammatory bowel diseases, oncogenesis, and innate immune responses to cancer. Additionally, deployment of the new Cre driver will shed light on factors that govern the functional plasticity of ILC3s in inflammatory settings.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Mental side effects of chemotherapy reduce quality of life and treatment dosing, thereby increasing mortality. Current supportive care treatments are ineffective, thus identifying new interventions to prevent or reduce these adverse side effects are crucial. We and others have recently hypothesized that gut dysbiosis may contribute to the behavioral side effects of chemotherapy. Our long-term goal is develop novel microbial-based treatments for the devastating side effects of the large and growing populations of chemotherapy-treated patients. The gut microbial community is dramatically altered by chemotherapy and has recently been shown to communicate with the brain to affect behavior. Indeed, we have previously published that altered composition of the gut microbiota due to chemotherapy in mice drives central and systemic inflammation resulting in selective behavioral side effects. Thus, the overall objective here is to determine the potential and feasibility of using fecal microbial trans- plant (FMT) as an intervention for chemotherapy behavioral side effects. Three specific aims are proposed to study this objective using our murine chemotherapy model (Aims 1 & 2) and a feasibility pilot clinical trial in breast cancer patients receiving chemotherapy (Aim 3). Aim 1 will quantify the efficacy of healthy FMTs in ameliorating sickness and cognitive behaviors during chemotherapy. Efficacy of autoFMT, alloFMT, or saline delivered in a preventative or therapeutic paradigm on sickness and cognitive behaviors will be assessed during chemotherapy treatment in mice after mammary tumor resection. Aim 2 will identify the systemic and neurobiological correlates of FMT interventions during chemotherapy. The efficacy of the FMT treatments on systemic and central inflam- matory signals, and the specific role of microglia, will be assessed during chemotherapy in mice after tumor resection. Aim 3 will pilot the feasibility of investigating the therapeutic potential for FMT to alleviate behavioral and gastrointestinal chemotherapy side effects in early-stage breast cancer patients. A small group of patients receiving chemotherapy will undergo an autoFMT or standard of care. Side effects will be assessed before and after FMT. The proposed research is conceptually innovative and our interdisciplinary team will be the first to use novel therapeutic FMT for chemotherapy-induced behavioral side effects in a combined basic and clinical approach. This research project will provide the necessary mechanistic and clinical foundation for future pro- posals focused on larger clinical FMT trials. This is the first step along a microbial research arc that has the potential to alter how common chemotherapy drugs are administered.

NIH Research Projects · FY 2026 · 2024-01

Project Summary Over the past 30 years, the synthesis of small molecules has become commonplace to facilitate drug discovery and development efforts. Retrosynthesis reduces a structurally complex target molecule into increasingly structurally simpler intermediates and commercially available starting materials, facilitating the preparation of a target molecule (also referred to as product) through a series of logical synthetic reactions (i.e., a multi-step synthetic route) from readily available starting materials or building blocks (referred to as reactants). Retrosynthetic analysis has become the cornerstone of modern synthetic endeavors and has revolutionized drug design. The goal of this project is to develop innovative generative AI – the type of AI that can create new content, to generate synthetic reaction libraries with diverse and feasible reactant molecules to synthesize given molecules via one-step synthetic reactions, and to evaluate and validate the libraries in laboratories thoroughly and rigorously. To achieve the goal, we have the following Aims. Aim 1 is to generate diverse and high-quality synthetic reaction libraries by developing innovative deep graph generative methods for retrosynthesis prediction. Novel graph neural networks will be developed to best capture and represent molecular structures for downstream retrosynthesis analysis. Innovative graph-based generative methods will automate step-by-step modification of target molecules toward their reactants. Aim 2 is to generate diverse and high-quality synthetic reaction libraries by developing innovative sequence-based methods for retrosynthesis prediction. Novel sequence-based methods include pre-training strategies, SMILES editing and a reinforcement learning framework will be developed. Aim 3 is to evaluate the generated synthetic reactions by domain expertise and laboratory experiments. Successful completion of this project will enable diverse and high-quality synthetic reaction libraries for any given drug-like molecule, which will be highly significant to accelerating drug development (e.g., lead generation). More importantly, the project will enable new AI capacity and infrastructure far beyond the conventional methodologies in synthetic route design. Successful application of the new methodology is ultimately expected to facilitate rapid retrosynthetic analysis of newly discovered or complex molecules as well as those with limited availability, enabling chemical synthesis for subsequent biological evaluations.

NIH Research Projects · FY 2026 · 2024-01

Project Summary My lab studies the molecular biology of archaeal microorganisms. Our rationale for doing so lies in the evolutionary relationship between archaea and eukaryotes. Archaeal possess information machineries that are in essence a simplified, ancestral version of the eukaryotic machinery. Viewing these machineries through the prism of archaea provides a unique perspective on the fundamentally conserved molecular mechanisms of genome duplication and expression. We have investigated these processes for many years and have in vitro systems in place for both DNA replication and gene transcription. More recently, we have discovered that archaea of the order Sulfolobales organize their genomes with a compartmentalized architecture – reminiscent of that seen in higher eukaryotes. Our work has revealed a key role for a simple, small SMC-superfamily protein, termed ClsN, in establishing and maintaining this compartmentalized architecture. We are now ideally poised to integrate our chromosome architecture studies with our DNA replication work, to investigate the interplay between genome organization and the dynamic processes of DNA replication and gene expression through combined in vitro, single-molecule and in vivo approaches. With regard to gene expression, we have recently described an unanticipated role for ClsN in effecting an epigenetic state in Sulfolobus. We will dissect the mechanistic basis of this state, determine how applicable it is to other regulatory systems in Sulfolobus and investigate the mechanistic basis of the multi- generational inheritance of ClsN-mediated conformational marks. Taken together, our work will reveal novel insights into the core molecular biology of archaea. Additionally, due to the relationship between archaea and eukaryotes, our work will provide a novel perspective on the evolution and core mechanisms of orthologous processes in eukaryotes.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT Randomized controlled trials (RCTs) are the gold standard for assessing interventions for preventing and treating cancer, but their external validity is only guaranteed if the trial participants are a random sample from the target population. Unfortunately, most cancer-related RCTs use convenience samples, not probability samples, and differences between the trial sample and the target population are likely to exist. If these differences are related to the effectiveness of the treatment being studied (“effect modifiers”), trial results will fail to generalize. While observable differences may be assessed and potentially adjusted for (e.g., underrepresentation of certain demographic groups), these differences have been shown to not completely explain the so-called efficacy-effectiveness gap. We posit that unmeasured differences between who chooses to participate in an RCT and who does not may be an important contributor to the failure of some trial results to generalize. In this project, we propose to develop a statistical framework for quantifying the potential impact of unmeasured differences between the trial sample and the target population on trial results. The resulting sensitivity analysis will bound the potential bias in the treatment effect estimate when generalizing from the trial sample to a target population. The methodology will be based on our prior work developing sensitivity analyses in the areas of survey nonresponse and selection bias which similarly consider the issue of differences between who is in a study sample and who is not. This work will have broad applicability beyond cancer trials, as generalizability is a universal concern of randomized trials across application areas.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT T-cell large granular lymphocytic leukemia (T-LGLL) is an incurable, underdiagnosed proliferation of clonal CD8+ cytotoxic T-lymphocytes that results in severe neutropenia and anemia, with resultant recurring infections, transfusion dependence, and death. There are no FDA-approved therapies for T-LGLL, and current immune- suppressive based therapies have marginal efficacy. Even if a response is attained with current therapies, patients shift between immune-suppressive agents, are subject to the adverse effects of these drugs, and for those with any effect, indefinite therapy is required. Urgent investigation into the pathogenesis of T-LGLL and development of rational, targeted therapies are needed. T-LGLL is a cytokine-dependent disease, driven by interleukin-15 (IL-15) which has been identified as the ‘master switch’ crucial to induce and potentiate T-LGLL. IL-15 induces pathogenesis in T-LGLL through up-regulation of STAT3, with resultant decrease in Fas/Fas- Ligand mediated apoptosis with resultant T-LGLL proliferation and cytopenias. We evaluated BNZ-1, a γc-inhibiting peptide that blocks IL-15, in patients with T-LGLL in only the second major, multicenter trial completed in T-LGLL. Clinical responses were observed in 20% of patients, but in vivo data revealed that nearly all patients had dramatic apoptosis of T-LGLL cells 24 hours post-BNZ-1, though apoptosis persisted only in responding patients at day 29. These results (in revision, Blood), provide in vivo proof that BNZ-1 induces T- LGLL cell apoptosis, and that T-LGLL cells are dependent on IL-15 in vivo in patients. To evaluate the mechanisms of resistance to IL-15 deprivation induced cell death, we performed single-cell RNA sequencing (scRNAseq) on samples from a non-responder. We identified the emergence of unique T-LGLL sub-populations on day 29, with corresponding up-regulation of anti-apoptotic pathways PI3K and NF-kB; implicating up- regulation of these alternate pathways in T-LGLL as key mechanisms of resistance of T-LGLL to IL-15 deprivation. Yet, validation in a larger sample set is needed to confirm these findings and identify therapeutic targets. To address these translational and clinical gaps, we will perform scRNAseq and single-cell T-cell receptor sequencing (scTCRseq) on remaining clinically annotated samples from 3 responders and 3 non- responders from the BNZ-1 trial to test our hypothesis that resistance to IL-15 deprivation with BNZ-1 is caused by expansion of resistant T-LGLL populations with upregulation of anti-apoptotic pathways (e.g. NF-kB, PI3K). In Aim 1, we will perform scRNAseq at serial timepoints (baseline, 24 hours post-BNZ-1, 29 days post BNZ-1), to identify key anti-apoptotic pathways, genes, and therapeutic targets in resistant populations. In Aim 2, we will apply scTCRseq to evaluate T-LGLL clonal populations (clonotypes), and determine the impact of IL-15 deprivation on gene expression and anti-apoptotic pathways in these groups. Upon completion, we will gain crucial mechanistic insights on the critical genes and anti-apoptotic pathways in T-LGLL that lead to resistance to IL-15 deprivation and identify therapeutic targets for a future NCI R01 proposal and BNZ-1 combinatorial trial.

NIH Research Projects · FY 2026 · 2024-01

Abstract Anthracycline-based chemotherapeutics such as doxorubicin (Adriamycin) are among the most widely used anticancer agents in oncology for the treatment of multiple solid tumors and leukemias. The clinical use of doxorubicin is associated with a dose-limiting, potentially lethal cardiotoxicity for which no effective preventative treatments are presently available. In addition, the mechanism by which doxorubicin accumulates into cardiomyocytes remains to this day unknown. Using a technique based on human induced pluripotent stem cell-derived cardiomyocytes from cancer patients receiving doxorubicin, we recently found that uptake transporter OCT3 is highly upregulated in patients experiencing cardiotoxicity. Functional validation studies in OCT3-deficient mice and heterologous overexpressed models confirmed that doxorubicin is transported into cardiomyocytes by OCT3. Furthermore, deficiency of OCT3 protected mice from acute and chronic doxorubicin-related changes in cardiovascular function and genetic pathways associated with cardiac damage, and these findings were confirmed using cardiac MRI-based methods. To provide proof-of-principle and demonstrate translational relevance of this transport mechanism, we found that pharmacological targeting of OCT3 can also preserve cardiovascular function following treatment with doxorubicin without affecting its plasma levels and its cytotoxic potential against multiple leukemia and breast cancer cell lines. Finally, we identified a previously unrecognized, OCT3-dependent pathway of doxorubicin-induced cardiotoxicity that results in a downstream signaling cascade involving the calcium binding proteins S100A8 and S100A9, and we validated this observation in a mouse model with S100A8 and S100A8 deficiency. Based on these preliminary findings, we now outline three sets of related studies that will further test and refine the validity of our central hypothesis that targeted inhibition of OCT3 function can specifically affect accumulation of doxorubicin in cardiomyocytes and affect downstream toxic events without negatively influencing its plasma pharmacokinetic profile or antitumor properties: (i) identification, validation, and mechanistic characterization of novel OCT3 inhibitors from a library screen that includes FDA-approved agents in novel humanized knock-in and conditional knock-out mouse models; (ii) functional validation of endogenous and exogenous cardiac-specific OCT3 biomarkers that could serve as a companion diagnostic to guide dose selection of OCT3 inhibitors; and (iii) safety, toxicokinetic, and efficacy analyses of optimized combinatorial regimens of OCT3 inhibitors with doxorubicin (acute and chronic), including simultaneous assessment of cardiac protection and antitumor properties in established experimental models of breast cancer and acute leukemias. It is expected that these studies will shed new light on the etiology of doxorubicin-induced cardiotoxicity and provide a rationale for the future implementation of novel targeted intervention strategies to prevent this debilitating side effect.

NIH Research Projects · FY 2025 · 2024-01

Project Summary/Abstract The focus of the Zadrozny laboratory is the design of metal complexes for noninvasive sensing of physiological chemistry. The broader goal of the effort is to make molecular probes that overcome inherent challenges in electron paramagnetic resonance imaging (EPRI), the unpaired electron analog to conventional 1H MRI. EPRI can sense local chemistry and could produce comprehensive chemical and anatomical maps of the body if merged with 1H MRI. Modern EPRI molecular imaging probes are organic radicals which require dangerous high-energy microwaves for use in the large magnetic field of an MRI scanner. Hence, the two techniques remain disconnected. For EPRI to enable imaging of physiological chemistry by integration with MRI, new probes must be developed to avoid high frequency microwaves at high magnetic fields. The next five years of the Zadrozny lab’s work involve exploring high-spin metal complexes as an alternative platform to radicals for molecular probes in EPRI. A key inherent advantage of metal ions is that the unique electronic feature of large zero-field splitting enables the possibility of safe, low-frequency microwave use at high magnetic field. Hence, metal complexes with this feature could provide a completely new set of EPRI molecular imaging probes with capabilities unmatched by organic radicals. However, all of the basic EPR spectral properties of metal complexes with low frequency microwaves are unmapped. The Zadrozny lab will amend this knowledge gap. The work will use synthetic inorganic chemistry and spectroscopic analyses to (1) understand how to target the frequency/field of the resonance to match the magnetic fields of MRI scanners with low-frequency microwaves (2) understand how to control the linewidth of the low-frequency EPR resonances to enhance resolution, and (3) how to merge radical/metal chemistry in hybrid molecules to gain the advantages of both metals and radicals for a single molecular probe system. Meeting these objectives will provide a new class of imaging probe capable of mapping physiological chemistry in a conventional MRI scanner.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY Tertiary lymphoid structures (TLS) are T and B lymphocytes in aggregation with antigen presenting cells, fibroblasts, and high endothelial venules observed in some chronically inflamed tissues. TLS formation in tumors of cancer patients is associated with improved adaptive immune function and increased overall survival. In surgically resected human pancreatic ductal adenocarcinoma (PDAC), a significant proportion of patients form spontaneous TLS in their primary tumors. Other groups have demonstrated similar findings, but no evidence currently exists showing TLS directly influence T cell immunity. In this application we propose to investigate if TLS enhance T cell immunity in the context of PDAC tumorigenesis and the mechanism of how this occurs. Our preliminary data in TLS+ mouse and human PDAC tumors shows TCF1+ stem-like T cells (T-stem) are enriched specifically within TLS relative to the surrounding tumor and stroma. We posit that TLS in PDAC tumors support the development of stem-like T cell phenotypes (T-stem), dependent on interleukin-21 signaling. These unique T cell subsets have previously been shown to sustain cellular immunity during chronic infection and cancer, providing continual repletion of memory, effector, and terminally differentiated cells following antigenic recall or immune checkpoint blockade treatment. We model this biology using a panel of mouse PDAC cell lines derived from transgenic mouse tumors that conditionally express mutant KRas and p53, the most common driver mutations in PDAC, and encode for a model MHCI-restricted antigen to track tumor antigen-specific responses. These cancer cell lines, despite sharing a similar genetic and mutational background, induce tumors with unique phenotypes and heterogenous tumor microenvironments. We will deploy a novel method to induce TLS in these mouse tumors through lymphotoxin beta receptor (LTBR) agonism, whereby some of our mouse tumor lines are permissive for TLS formation and others are resistant. We will test if the TLS-mediated generation of T-stem is responsible for improved tumor control in CD8 conditional TCF1-knockout mice. We will determine if their enrichment in TLS is due to IL-21 signals derived directly from TLS that promote their formation, independent of lymphatic involvement, utilizing a combination of cell lineage tracing and genetically modified mouse models. We will complement these studies by investigating tumor samples from PDAC patients for phenotypic pathways unique to specific regions of these tumors. We will achieve this by performing spatial transcriptomics on human PDAC tumor sections analyzed by an innovative algorithm that transforms spatial variable genes into functional tissue modules to determine region-specific immune phenotypes localized to the TLS compared to non-TLS regions and tumors without TLS. This approach identifies the mechanism of how TLS supports T cell function in authentic mouse models of TLS+ PDAC and translates these findings to human cancer patients so novel immunotherapy options designed to elicit and activate TLS can be offered to patients with a treatment resistant malignancy.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY/ABSTRACT Failure of the right ventricle (RV) is a main component of the morbidity and mortality of pulmonary hypertension (PH). There exists no specific treatment for RV dysfunction and failure and the only cure for many patients (especially in the case of pulmonary arterial hypertension) remains transplantation. This is partially due to the critical lack of understanding of RV biology; therapeutic strategies that are beneficial for LV failure have worse outcomes when applied to those with RV failure. What has been observed in RV dysfunction in human subjects and animal models is exaggerated fibrosis and divergent contractile and hypertrophic responses to several drugs when compared to LV dysfunction. Thus, the RV may contain distinct pathobiology compared to the LV that has high significance for treatment of PH. A greater understanding of RV cellular biology is needed to develop new opportunities toward our long-term goal of discovering tailored therapies for dysfunction and failure of the RV. We performed transcriptomic analysis to compare the RV with the LV using Sprague Dawley rats and found significant differences in immune-related and fibrotic genes. Flow cytometry was used to confirm that the RV of control rats had an average of four-fold higher density of immune cells, namely macrophages/monocytes, compared to the LV. When rats were subject to pressure overload via hypoxia in combination with the VEGF pathway inhibitor Sugen, the RV became hyper-fibrotic. This was also clear in isolated fibroblasts from the RV of control rats, which had a higher proliferation rate and an alteration of metabolic genes. Thus, we were led to our central hypothesis: the RV is primed for disproportionate dysfunction under pressure overload due to the unique physiology of its cellular constituents, namely immune cells and fibroblasts. We will investigate this hypothesis with the following two aims: Aim 1: Determine the role of the immune response in the RV during adaptive and adverse remodeling. We will test the hypothesis that distinctive macrophage populations in the RV are critical for the “hyper-fibrosis” found in RV disease, and modulating specific inflammatory signals will be able to dampen the fibrotic response. Aim 2: Identify the fibrotic mediators in RV fibroblasts in situ and in vitro. We will test the hypothesis that RV fibroblasts have altered cellular metabolic programming that causes an enhanced fibrotic response, mediated by interactions with cardiac macrophages. This project will provide new understanding of the cellular processes that occur during pathologic remodeling of the RV. This knowledge is expected to yield new therapeutic approaches for RV dysfunction and failure occurring with PH, which currently have no specific treatment.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY There are ~1 billion human influenza cases worldwide each year, resulting in up to 650,000 deaths. Influenza virus infects cells of the airways and distal lung, causing acute lung injury. The main class of antiviral drugs recommended for treating influenza works by blocking the viral neuraminidase enzyme on the virus surface to prevent the release of viral particles from infected cells. However, the increasing emergence of drug resistance limits their effectiveness. Therefore, there is an urgent need to identify novel molecular regulators and pathways of the innate immune response in the lung against influenza virus infection. This proposal is driven by our novel finding that the E3 ubiquitin ligase MARCH10, a protein never described in the lungs in the context of influenza virus infection with no known role in pulmonary immunity, is significantly downregulated by influenza virus infection in human and murine lungs. Overexpression of MARCH10 also drastically decreases the viral protein influenza hemagglutinin when lung epithelial cells are infected with influenza virus. We hypothesize that MARCH10, regulated at the transcriptional level, decreases influenza pathogenesis and is a critical regulator of airway epithelial cell host defense. We have proposed the following specific aims to investigate this hypothesis: 1) Determine if influenza virus induces a stage-specific decrease in MARCH10 at the gene transcriptional level and 2) Determine if catalytically active MARCH10 reduces influenza virus pathogenesis via post-transcriptional mechanisms. To accomplish these goals, the principal investigator has developed a five-year training program under the guidance of her mentors, Dr. Rama Mallampalli – an expert in ubiquitin-mediated proteolysis and acute lung injury and Dr. Jacob Yount - an expert in influenza and viral immunology, to acquire training in advanced molecular and cell biology techniques, virology techniques, and small animal model development. The candidate’s training will also be overseen by a Scientific Advisory Committee to lend expertise, provide oversight and evaluate progress, including, Dr. Mallampalli, Dr. Yount, Dr. Estelle Cormet-Boyaka – an expert in lung epithelial cell biology, and Dr. Dan Wozniak – an expert in lung and bacterial pathogenesis. The proposed career development plan will provide the additional training necessary to achieve the principal investigator’s ultimate goal of becoming an independent physician-scientist studying the biological mechanisms by which lung epithelial cells respond to respiratory viral infections and modulate the lung innate immune response and translate these findings into novel therapeutics to prevent and treat viral infections and its associated lung injury.

NIH Research Projects · FY 2025 · 2023-12

PROJECT SUMMARY Cancer metabolic reprogramming is key for the adaptation of the tumors to their harsh microenvironments. Elevated LD abundance in tumors is frequently detected and is being recognized as a common feature of their rewired metabolism. Lipid droplets are multifaceted organelles that participate in crucial cellular functions including metabolism, membrane synthesis and homeostasis, signaling, and protein trafficking. We and others have shown that hypoxia stimulates LD accumulation and that forced depletion of LD stores impedes tumor growth. In order to discover novel regulators of lipid droplet turnover, we performed a genome-wide loss of function screen based on Crispr engineering. Edited cells were separated based on their LD abundance and sequenced. Our approach identified GFPT1 (glutamine-fructose-6-phosphate transaminase 1) as a candidate positive LD regulator. GFPT1 catalyzes the first and rate limiting step in hexosamine biosynthesis. The hexosamine biosynthetic pathway uses a glycolytic intermediate and glutamine to produce UDP-GlcNAc which is essential for O-GlcNACylation and glycosylation of proteins and other macromolecules. Flux through the hexosamine pathway is regulated by oncogenic events, nutrient availability, and input from nutrient sensors like AMPK. In turn, HBP impacts crucial signaling functions and metabolic processes. Cancers have elevated HBP activity and gene expression and are dependent on this activity for survival, growth, and metastatic dissemination. Broad range pharmacological inhibitors of the HBP have shown promising results as a therapeutic approach, however, there are no HBP-directed cancer therapies currently approved. Our preliminary finding that GFPT1 positively regulates lipid storage prompts us to hypothesize that concurrent inhibition of the HBP and neutral lipid synthesis will result in more complete LD depletion and enhanced tumor control. In aim 1 we will delineate the lipid metabolic processes impacted by genetic GFPT1 ablation or small molecule inhibition of HBP in PDAC models in vitro. We will uncover nutritional requirements for LD turnover under normoxia and hypoxia in the context of HBP inhibition and will quantify the toxicity of combined HBP- and lipid metabolism inhibition. Further, we will mechanistically link GFPT1 and AMPK via post-translational modifications during normoxic and hypoxic LD accumulation. In aim 2 we will evaluate the in vivo efficacy of HBP and metabolic inhibitors in orthotopic PDAC models and will measure the impact of these new treatment schemes on tumor microenvironmental characteristics. Successful completion of this proposal will uncover a new connection between two cancer-promoting metabolic programs and will develop new chemotherapeutic paradigms for cancer treatment.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY Developing neuroprotective and repair strategies represents an urgent unmet need for treating inflammatory demyelinating disorders of the central nervous system (CNS) including multiple sclerosis (MS). Lesions in both MS patients and the MS mouse model, experimental autoimmune encephalomyelitis (EAE), involve T lymphocyte infiltration and axonal damage. However, interactions between CNS-infiltrating T cells and central neurons remain poorly understood. 4-Aminopyridine (4-AP) is an FDA-approved drug for the symptomatic treatment of MS to improve walking speed. Although it is commonly believed that 4-AP blocks Kv1 (Shaker) voltage-gated K+ (Kv) channels to enhance axonal conduction and suppress immune response, the mechanisms underlying 4-AP’s actions in treating MS are still not completely clear. Our new results have provided several lines of compelling evidence that a Kv3 (Shaw) channel, with high 4-AP sensitivity, is expressed in both CD4+ T cells and CNS neurons, and plays a key role in T cell-induced axonal injury. This channel regulates T cell activation that is required for EAE induction, shown by our active and passive EAE results using its global knockout (KO) mice. This notion is further supported by our data from T cell culture, immunostaining/confocal imaging, flow cytometry, RNAi knockdown, and conditional KO (cKO) mice. Therefore, based on our new findings, we propose a novel hypothesis that in the development and pathology of CNS autoimmunity, Kv3 channel is required for the efficient generation of encephalitogenic T cells, whereas its upregulation in axons aggravates autoimmune-induced injury via aberrant Ca2+ signaling. To test this original hypothesis, we have created a floxed mouse line for this Kv3 channel to examine the effects of its cell- type-specific cKOs on EAE. We will use a multidisciplinary approach, including active and passive EAE models, inducible cKO and transgenic mouse lines, flow cytometry, confocal and transmission electron microscopy, functional assays, in vivo RNAi, and a new myelin coculture. We will determine (Aim 1) whether deleting this Kv3 channel from CD4+ T cells suppresses EAE induction and progression via impaired T cell activation, (Aim 2) whether deleting this channel from CNS neurons ameliorates autoimmune-induced axonal injury, and (Aim 3) how this channel regulates pathogenic interactions between T cells and axons in autoimmune-induced injury. This project is innovative because this is the first study to show that a Kv channel regulates the function of both immune cells and neurons. This project is significant because the findings of this project will provide novel mechanistic insights into pathogenic interactions between T cells and neurons, and hence may contribute to a novel treatment strategy for MS through simultaneously suppressing immune response and rescuing injured axons.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY/ABSTRACT Social cognitive deficits are prevalent in schizophrenia spectrum disorders (SSD) and significantly contribute to poor community functioning. Since most people living with SSD are in their mid- and late-life and social cognitive deficits often persist or even worsen over the course of illness, there is a critical need to develop effective interventions for mid-late life SSD. Characterizing social cognitive deficits and their changes across age and identifying neural markers are prerequisites to developing efficient identification strategies and targeted neurobiological treatments. In response to RFA-MH-22-270 “Schizophrenia and Related Disorders during Mid- to Late-life,” this project aims to advance knowledge in social cognition—one of the identified priority research areas. We will recruit a large sample (n = 192; 50% female) of SSD participants in their mid- late life (age 35 to 75). Additionally, 48 early-psychosis (age 18-34) and 120 age-matched non-psychiatric participants will serve as clinical and healthy comparisons, respectively. Participants will complete assessments of psychiatric phenotypes, neurocognition, and community functioning. Social cognition will be assessed using a comprehensive battery capturing low- to high-level processes. A subset (75%) of the participants will additionally undergo EEG during social cognitive tasks to determine the theta-band neural oscillatory features underlying social cognitive deficits. The specific aims of this project are threefold: 1) Delineate the age trajectories of social cognitive deficits in mid-late life SSD; 2) Evaluate theta-band neural oscillatory features as neural markers of social cognitive deficits in mid-late life SSD; and 3) Parse heterogeneity based on neural oscillatory signatures in mid-late life SSD. Successfully completing these specific aims will advance our understanding of the course and neural mechanisms of social cognition in mid- late life SSD, advancing NIMH’s Strategic Objective Objectives 2.1 (characterize the trajectories of cognitive and affective processes across the lifespan), 2.2 (identify behavioral and biological markers of mental illnesses), and 1.3 (identify neural mechanisms contributing to mental illnesses). The findings will guide identification and personalized treatment strategies, providing critical knowledge to determine who and how to intervene.

NIH Research Projects · FY 2026 · 2023-12

Project Summary/Abstract Conventional thresholds of physiological parameters for clinical diagnosis have become a root cause of racial/ethnic disparities in healthcare. These thresholds were established decades ago using limited samples of Caucasian subjects and simplistic statistics without considering genetic effects. Consequently, these thresholds have no capacity to account for the normal variability intrinsic in race/ethnicity, genes, and environments, which can lead to errors in disease diagnosis and risk assessment when applied to racial/ethnic minorities. As the minority population grows and becomes even more diverse, continuing to use Caucasian-based, “one-size-fits- all” thresholds creates the risk of exacerbating racial/ethnic disparities in healthcare. Therefore, an effective alternative must be developed to address this issue. Hence, we propose creating Person-specific Precision Thresholds (PPTs), which are designed to account for normal variability in genetic and biological makeup, variation in race/ethnicity, sex, age, and other characteristics. PPTs will eliminate the root cause of healthcare disparities caused by conventional thresholds for diagnosis and risk assessment. The innovative precision of PPTs will fundamentally shift current research paradigms regarding clinical thresholds from a single cutoff point for everyone to precise, person-specific thresholds matched to individual genetics and phenotypic characteristics. PPTs will effectively improve disease diagnosis and risk assessment while reducing healthcare disparities. The first PPT iteration will focus on women's bone mineral density (BMD), as osteoporosis primarily affects women the most. The existing T-score threshold of BMD has become controversial because many women who sustain fragility fractures have a “normal” BMD value, as defined by the commonly employed T-score method. Because genetic factors contribute to 50-85% of BMD variation, genetic variants must be considered when creating precision thresholds. With feasibility tested by pilot studies, we hypothesize that PPTs will enhance the ability of a single BMD measure to predict fractures in minority women compared to conventional T-score and prior model-based methods, which lack consideration of genetics. We will pursue the following three specific aims: (1) Develop a modeling framework for PPTs of BMD using novel machine-learning techniques; (2) Calibrate PPTs of BMD using clinical data; (3) Validate the PPTs of BMD using genomic cohort data. In addition, we will create a publicly available, cloud-enabled PPT application for community investigators. This proposed approach is innovative because it integrates individual genetics into clinical precision thresholds and uses novel machine- learning techniques to create PPTs. By replacing the traditional one-size-fits-all threshold, the innovative PPTs will shift existing research paradigms regarding how clinical thresholds have been developed and implemented. The knowledge gained from this research can be utilized to create many other PPTs to improve the diagnosis and prevention of various diseases and illnesses. These next-generation PPTs will help significantly improve patient outcomes, leading to better health, a higher quality of life, and improved health equity.

NIH Research Projects · FY 2025 · 2023-12

Contact PD/PI: Vilgelm, Anna E Project Summary Metastatic melanomas are aggressive tumors that respond poorly to chemotherapy and radiation and promptly acquire resistance to targeted therapy. While immune checkpoint inhibitors (ICIs) can induce sustained melanoma remission, nearly half of patients are intrinsically resistant to ICIs, in part because their tumors are too large to be cleared by the immune system or are immunologically “cold” and lack immune recognition necessary to generate anti-tumor T effector cells. Our preliminary data show that senescence- inducing therapy enhances melanoma-cell intrinsic pro-inflammatory program driven by transcription factor NF- κB. We identified the RIG-I-Like Receptor (RLR) pathway as a candidate NF-κB inducer. This pro-inflammatory program involves enhanced antigen presentation and secretion of chemokines that recruit antigen-presenting cells into the tumor. We also found that senescent cells overexpress checkpoint molecules PD-L1 and Galectin 9, which can inhibit T cells by binding their inhibitory receptors PD-1 and TIM-3, respectively. This suggests that senescence-inducing therapy can enhance recognition of tumor antigens while targeting senescence- specific immune checkpoints can promote adoptive T cell responses against these antigens. Despite its immunogenic properties, therapeutic application of senescence-inducing agents is complicated by the reports that persistent senescent cells may promote tumor metastasis, relapse, and chemo-resistance. Here we hypothesize that we can safely improve melanoma immunogenicity by inducing tumor cell senescence for a limited time, followed by elimination of persistent senescent cells using senolytic drugs that specifically kill senescent cells. This novel therapeutic approach is expected to be effective by: 1) directly blocking tumor cell growth and survival, 2) activating RLR-mediated tumor cell intrinsic immuno-stimulatory signals, and 3) making tumor cells vulnerable to inhibitors of PD-L1/PD1 and Galectin 9/TIM3 immune checkpoints. Three specific aims will examine these mechanisms. In the first aim we will establish the optimal dosing schedule for combined senescence-inducing and senolytic therapy and determine the direct anti-tumor efficacy in an immune-deficient mouse model. A pre-clinical trial using our extensively characterized melanoma PDXs will be conducted to estimate response rate to senescence-inducing and senolytic drug combinations and to identify response-predictive biomarkers. Second aim will investigate the mechanisms whereby senescence-inducing and senolytic therapy facilitates the innate immune sensing of tumor focusing on the role of RLRs. We will examine whether deficiency in RLR signaling can abrogate NF-κB activation, chemokine secretion, and immune cell recruitment into senescent tumors. Aim three will determine if senescence-inducing and senolytic therapy augments immune checkpoint inhibitor (ICI) responses. We will utilize anti-PD-L1 and anti-TIM3 agents to target senescence-cell specific immune checkpoints. These studies will provide a preclinical basis for clinical development of rational combinations of senescence-inducing and senolytic agents. Page 6 Project Summary/Abstract

NIH Research Projects · FY 2026 · 2023-12

Project Summary/Abstract Myocardial infarction (MI) is a major cause of death and disability worldwide, affecting >800,000 Americans annually. A delicate balance of repair and remodeling processes is required to preserve the structural integrity and reinforce the injured myocardium following MI. Cardiac fibroblasts (CFs) are among the most numerous cell types in the heart and play an essential role in fibrotic remodeling. Following ischemic injury, surviving CFs exhibit a highly dynamic response involving the transition into an activated phenotype characterized by increased proliferation, migration to the infarct region, and secretion of fibrotic proteins. Further, CFs secrete a battery of paracrine signals to help coordinate the organ-level response to injury. Thus, proper healing requires dynamic spatial and temporal control over CF function. Dysregulation of the CF response to injury promotes pathological fibrosis, increased risk for arrhythmia, and cardiac dysfunction. While there have been many studies exploring the diverse signaling cascades and stressors that cause CF activation, the precise molecular pathways responsible for orchestrating dynamic changes in CF function across spatial and temporal scales are not well understood. Spectrin proteins are important for providing structural membrane support and spatiotemporal regulation for cell signaling events. Recent work identified stress-induced loss of βIV-spectrin, to be an important step in CF activation and fibrosis. Further, loss of βIV-spectrin was found to depend on Ca2+/calmodulin-dependent protein kinase II (CaMKII). A broader role has been identified for βIV-spectrin/CaMKII in regulating CF gene expression through an interaction with signal transducer and activation of transcription 3 (STAT3), a signaling molecule and transcription factor that promotes profibrotic mechanisms. Specifically, CaMKII is activated and promotes loss of βIV-spectrin and redistribution of STAT3 to the nucleus that lead to changes in fibrotic gene expression. Further, we identified a molecular switch for βIV-spectrin stability involving direct phosphorylation by calmodulin- dependent protein kinase II (CaMKII). Together these data support our central hypothesis that βIV-spectrin coordinates a tunable network for temporal and spatial control of CF function and the normal healing process following MI. These studies seek to offer new mechanistic insight into how the myocardial repair process is orchestrated and to improve therapeutic options for patients who have experienced MI.

NSF Awards · FY 2023 · 2023-10

The project, “Collaborative Research: RUI: Trust but Verify: The Use of Intuition in Engineering Problem Solving” is a three-year study about engineering intuition conducted at Bucknell University, Arizona State University, and Embry Riddle Aeronautical University. The goal of this project is to examine the application of intuition by engineering practitioners and generate knowledge that promotes the professional formation of engineers and development of a stronger engineering workforce. Engineering intuition is a problem-solving skill developed through experience that engineering practitioners subconsciously use when under pressure from constraints (e.g., lack of time). Engineering practitioners regularly use and develop intuition on-the-job; however, the use of intuition is often discouraged in undergraduate education. Research questions in this study focus on the application of intuition in engineering problem solving, solving “ill” versus “well” structured engineering problems, and prior engineering experience levels when engineers solve structured problems. The study will use a variety of methods and techniques (such as Simulation Interviews). New engineering practitioners (senior undergraduates and those with less than one year of experience) and mid-career practitioners (6-10 years of experience) will participate. The knowledge generated from this study will provide a foundation for bridging the disconnect between classroom and real-world engineering practices, designing educational interventions that promote intuition development, and understanding how early intuition development can help level the playing field for all students regardless of individual background, including past engineering experiences. The project, “Collaborative Research: RUI: Trust but Verify: The Use of Intuition in Engineering Problem Solving” is a three-year study about engineering intuition conducted at Bucknell University, Arizona State University, and Embry Riddle Aeronautical University. The goal of this project is to examine the application of intuition by engineering practitioners and generate knowledge that promotes the professional formation of engineers and development of a stronger engineering workforce. Engineering intuition is a problem-solving skill developed through experience that engineering practitioners subconsciously use when under pressure from constraints (e.g., lack of time). Expertise in a specific area is a prerequisite of intuition and is a defining characteristic of the expert. Engineering practitioners regularly use and develop intuition on-the-job; however, the use of intuition is often discouraged in undergraduate education. The disconnect between intuition’s use in engineering practice and in engineering education, coupled with limited knowledge of the relationship between intuition, expertise, and experience, presents an important gap in our existing understanding of engineering problem solving and future workforce preparation. A qualitative approach will be employed to investigate four research questions: RQ1: How does the application of intuition manifest in engineering problem solving? RQ2: How does the application of intuition vary when approaching “ill” versus “well” structured engineering problems? RQ3: How does the domain of practitioner expertise influence the application of intuition when approaching “ill” versus “well” structured engineering problems? RQ4: How does prior engineering experience influence the application of intuition when approaching “ill” versus “well” structured engineering problems? Cognitive Task Analysis (CTA) will elicit expert knowledge from a sample of new engineering practitioners (senior undergraduates and those with less than one year of experience) and mid-career practitioners (6-10 years of experience). Per best practices, CTA methods (Simulation Interviews, Critical Decision Method, and Knowledge Audit Method) will be used to support robust data collection. The knowledge generated from this study will provide a foundation for bridging the disconnect between classroom and real-world engineering practices, designing educational interventions that promote intuition development, and understanding how early intuition development can help level the playing field for all students regardless of individual background, including past engineering experiences. 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 · 2023-09

Homelessness severely affects health and well-being and is particularly negative for youth. Between 70-95% of youth experiencing homelessness (YEH) report problem substance use and 66-89% have a mental health disorder. Youth appear to be at greater risk while living on the streets or being homeless than adults and are more vulnerable to long term consequences of homelessness. Multiple factors are uniquely associated with homelessness, driving substance use and adverse mental health consequences. However, limited research has identified pragmatic interventions that have a long-term ameliorating impact on the complex, multi-symptomatic issues among these youth. This study overcomes prior gaps in research through testing a multi-component comprehensive prevention intervention targeting an array of factors that may affect health indicators and longer-term health outcomes. In partnership with Star House, a drop-in center for YEH, youth between the ages of 14 to 24 years, will be engaged and randomly assigned to conditions using a dismantling design so that essential intervention components can be efficiently identified. In particular, youth (N = 300) will be randomly assigned to a) Motivational Interviewing/Community Reinforcement Approach + Services as Usual (MI/CRA + SAU, n = 80), b) Strengths-Based Outreach and Advocacy + SAU (SBOA + SAU, n = 80), c) MI/CRA + SBOA + SAU (n = 80) or d) SAU (n=60) through the drop-in center. In order to assess the longer-term prevention effects on substance use, mental health and other outcomes, all youth will be assessed at baseline and at 3, 6, 12, 18 and 24-months post-baseline. The primary goal of this study is to establish the impact of a comprehensive intervention embedded within a system that serves YEH, a community drop-in center, on youth’s opioid misuse and disorder, other substance misuse and disorders, mental health diagnoses, and other targeted outcomes. This study will offer unique information on pathways underlying change along with cost estimates to inform future implementation efforts in drop-in centers around the country. This study is part of the NIH’s Helping to End Addiction Long-term (HEAL) initiative to speed scientific solutions to the national opioid public health crisis. The NIH HEAL Initiative bolsters research across NIH to improve treatment for opioid misuse and addiction.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY Age-related macular degeneration (AMD) is the third leading cause of blindness in the US and one of the leading causes of blindness worldwide. Initially, dry AMD results from inflammation caused by excess reactive oxygen species (ROS), and has no cure. When it progresses to wet AMD, the disease is characterized by abnormal growth of leaky blood vessels caused by excess expression of vascular endothelial growth factor (VEGF), which permanently damages the retina and causes severe vision loss. Anti-VEGF therapeutics are the current standard treatment for wet AMD. However, the requirement of frequent intravitreal injections is associated with high treatment costs, patient burden, and risk of complications including pain. This also does not address the inflammatory component of early stages of the disease. There is a clinical need to reduce injection frequency and treat underlying inflammation causing the disease. The overall objective of this project is to develop and validate an injectable, biodegradable, stimuli-responsive nanoparticle delivery system that can sustain release of a new therapeutic to treat inflammation for several months. In Aim 1, an investigational therapeutic will be synthesized and characterized. Stimuli-responsive polydopamine nanoparticles that release more therapeutic in the presence of reactive oxygen species (ROS) will be synthesized and loaded with the new therapeutic. Therapeutic and nanoparticles will be evaluated for in vitro cellular uptake in human retinal pigment epithelial cells using flow cytometry. Short-term biocompatibility of both the therapeutic and nanoparticles will be evaluated in vivo in a mouse model over 2 weeks. In Aim 2, in vivo biocompatibility and efficacy of the proposed treatments will be evaluated in the sodium iodate mouse model of dry AMD over 2 months. Structural and functional assessments of the eye will include intraocular pressure, fundus imaging, spectral domain optical coherence tomography, histology, and immunohistochemistry. Biocompatibility and biodegradation will be assessed concurrently. Therapeutic concentrations in ocular tissues will be validated by ELISA after 2 months. ELISA will also be used to evaluate retinal expression levels of HO-1 in the disease model to investigate the mechanism of action of the proposed therapeutic. These aims will evaluate a ROS-responsive drug delivery system to sustain release of an anti-inflammatory therapeutic in the eye. This has potential to treat underlying disease pathways associated with AMD and reduce intravitreal injections, improving the quality of life for patients with AMD and other ocular or inflammatory diseases.