Ohio State University

NIH Research Projects · FY 2026 · 2023-04

Project Summary: The pathogenesis of heart failure is linked to systemic metabolic dysfunction, a comorbidity likely involving peripheral organs, and resulting in insulin resistance in patients with and without diabetes. This study focuses on the reciprocal response of adipose tissue to pathological stress on the heart and will investigate how adipose tissue activity affects cardiac metabolic remodeling. This proposal originates from novel findings by each of the MPI laboratories. We have identified transient changes in adipose tissue plasticity during the pathogenesis of decompensated cardiac hypertrophy in response to pathological stress alone, absent any primary metabolic stress, such as diabetes, nutrient overload, or obesity. This finding and our unpublished data implicate cardiac natriuretic peptides (NP) as mediators of adipose activation and show that NPs are induced by a reduction in long chain fatty acid (LCFA) oxidation by the heart, even without pathological stress. Preliminary data reveal a profound, sex-specific array of early responses in white (WAT) and brown (BAT) adipose tissue activity to cardiac pressure overload that include beiging of WAT and induction of a lipolytic phenotype with cold exposure. While the cardiac responses to pathological stress include maladaptive remodeling of lipid metabolism, the effects of adipose plasticity on the cardiac lipid profile during the development of pathological hypertrophy are virtually unexplored. Thus, we hypothesize that: 1) the metabolic response of reduced fatty acid oxidation in the heart during pathological stress induces plasticity of WAT that is mediated by cardiac NPs in a sex-specific manner, distinct from β-adrenergic stimulation, with downstream effects on glucose tolerance; and 2) sustained beiging of WAT and activation of BAT confer cardioprotection against adverse cardiac metabolic remodeling. The research design supports three specific aims: 1. Elucidate effects of adipocyte adrenergic activation on WAT beiging, and responses of cardiac LCFA metabolism during TAC with adipose adrenergic activation and inhibition; 2. Determine whether adipose plasticity results from NP effects in direct response to TAC or from the metabolic shift of reduced LCFA oxidation in male and female mice; 3. Investigate a potential cardioprotective role of adipose browning on the lipid profile of failing hearts. The overall objectives are to: 1) determine the reciprocal metabolic responses between the pathologically stressed heart and adipose tissue via cardiac NP production and adipokine/lipokine release (namely 12,13-diHOME), respectively; 2) elucidate the consequences of sex differences in the responses of adipose tissue and myocardium to cardiac stress; and 3) explore the potential for WAT beiging or BAT activation to provide cardioprotection via metabolic signaling. The findings will contribute new insights into the adipose responses and contributions to metabolic remodeling of the heart during the pathogenesis of decompensated cardiac hypertrophy, with the potential to identify targets for remediating the progression to overt heart failure and improving peripheral and systemic metabolic dysfunction.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Pancreatic ductal adenocarcinoma (PDAC) is projected to become the second leading cause of cancer-related death by 2030, yet there are no accurate diagnostic tests for early diagnosis. Among pancreatic cystic lesions (PCLs), branch duct (BD) intraductal papillary mucinous neoplasm (IPMN) is the most common precursor lesion for pancreatic cancer. Nearly 50% of all prevalent cysts are BD-IPMNs. Endoscopic ultrasound (EUS)- guided fine needle aspiration (FNA) of PCLs and cyst fluid analysis are standard-of-care (SOC) diagnostic modalities. Unfortunately, the current SOC is suboptimal (65-75% accuracy) for the detection and risk stratification [high-grade dysplasia or adenocarcinoma (HGD-Ca) vs. low-grade dysplasia (LGD)] of BD-IPMNs. The goal of surgery in BD-IPMNs is to resect lesions with HGD-Ca. However, multiple surgical series over the last 5 years have revealed that nearly half to two-thirds of resected BD-IPMNs had only LGD, often representing overtreatment. In these instances, the morbidity (30%) and mortality (2%) from surgical resection of PCLs are not justified. On the other hand, several series reports missed (mean 13%) invasive cancers in BD-IPMNs during follow-up. There are currently no accurate tests for detecting HGD-Ca in BD-IPMNs. We have utilized a novel diagnostic modality of EUS-guided needle-based confocal laser endomicroscopy (nCLE), a technology that provides in vivo, real-time, optical biopsies of PCLs. In a landmark study, we demonstrated a high accuracy (97%) for nCLE-guided diagnosis of precancerous (includes mucinous BD-IPMNs) PCLs. We have derived nCLE features of HGD-Ca that can be qualitatively assessed and quantitatively analyzed in BD- IPMNs. We also have designed a pilot CLE-based convolutional neural network (CNN)-artificial intelligence (AI) algorithm to risk-stratify BD-IPMNs (HGD-Ca vs. LGD). We have also pioneered cyst fluid Next-Generation Sequencing (NGS) analysis, augmenting the diagnosis and risk-stratification of BD-IPMNs. The primary objective of the proposed study is to accurately risk-stratify (HGD-Ca vs. LGD) BD-IPMNs to detect early-stage PDAC and avoid unjustified pancreatic surgery. Supported by preliminary data, our central hypothesis is that EUS-nCLE (manual and CNN-AI algorithm) and a combination of EUS-nCLE with NGS and SOC variables will accurately risk-stratify BD-IPMNs. Specific aims – (1) Evaluate the accuracy and interobserver agreement of EUS-nCLE differentiation (HGD-Ca vs. LGD) of BD-IPMNs among independent observers. (2) Improve and prospectively evaluate an accurate nCLE-based CNN-AI algorithm for presurgical risk stratification (HGD-Ca vs. LGD) of BD-IPMNs. (3) Evaluate an integrative diagnostic approach including nCLE, NGS, and SOC to improve the accuracy of risk stratification (HGD-Ca vs. LGD) of BD-IPMNs. Successful completion of this project and application in clinical practice will provide a method for early detection of PDAC arising from PCLs, guiding surgical decision-making to help avoid unwarranted resections or delayed treatment.

NIH Research Projects · FY 2026 · 2023-04

My overall goal is to improve the health and well-being of individuals with comorbid opioid misuse/opioid use disorder (OUD) and chronic pain across varied settings and populations. Research on the management of comorbid opioid misuse/OUD and chronic pain is being outpaced by the clinical need across populations and settings. Additionally, there is no well-worn training pathway to conduct research in this area, few mid-career and senior investigators have the expertise to mentor early-career investigators in this space, and the need for mentoring to grow the workforce is enormous. I am a physician and PhD-trained behavioral scientist who treats individuals with opioid misuse/OUD and chronic pain. I have a consistent track record of NIH-funded research and productive mentoring at the intersection of these conditions across populations and settings including HIV, cancer, and primary care, and am at an institution with exceptional resources to support trainees. This K24 mid-career investigator award will allow me to grow the workforce at the intersection of opioid misuse/OUD and chronic pain by 1) expanding my capacity to mentor PhD students, postdoctoral scholars, and faculty from a variety of disciplines in research on comorbid opioid misuse/OUD and chronic pain; 2) Become a leader in implementation science approaches to studying treatment of comorbid opioid misuse/OUD and chronic pain, including hybrid trials; and 3) Develop the skills needed to systematically incorporate health policy considerations and methods throughout my work. I have assembled a team of advisors with whom I have created a comprehensive career development plan including didactic coursework, experiential learning, and conference attendance that will allow me to excel at achieving these goals. Additionally, this K24 proposes a new research project that leverages existing Delphi-based preliminary data to 1) Develop an implementation strategy bundle to promote the adoption and application of evidence-informed approaches (EIAs) to managing opioid misuse/OUD and metastatic cancer-related pain, and 2) Conduct a hybrid type 2 pilot trial of the approaches and implementation strategy bundle. Findings from this study will lead to a hybrid type 2 randomized trial of the implementation strategy bundle and ultimately improved care for patients with opioid misuse/OUD and metastatic cancer. This and other NIH- funded studies in my portfolio will provide mentees ample opportunity to propose mentored research projects. This mentored research experience, combined with a program of tailored career development opportunities, will develop a new cadre of independent investigators at the intersection of opioid misuse/OUD and chronic pain.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY In heart disease, the amount of blood pumped by the heart is not sufficient to supply the metabolic demands of the body. Current positive inotropes (dobutamine, milrinone) increase cardiac contraction by elevating the intracellular calcium contractile signal, however trials have demonstrated this elevated intracellular calcium also causes detrimental arrhythmia, impaired relaxation, and increased mortality rendering this inotropic approach unsuccessful as a long-term therapy. There are currently no approved therapies that directly increase the insufficient function of the heart in disease without long-term detrimental effects. The myofilament protein troponin I (TnI) is critical to relay the calcium activating signal into muscle contraction and is therefore a key regulator of in vivo cardiac muscle function. We have shown the site-specific phosphorylation of TnI increases cardiac muscle contraction without elevating intracellular calcium. Our preliminary data now demonstrates this TnI phosphorylation increases in vivo cardiac function in both normal and diseased hearts without long-term detrimental effects. This proposal will establish the novel effects and mechanism of this TnI site-specific phosphorylation to improve in vivo cardiac function in the normal and diseased mouse heart. We will further establish the effects and safety of this site-specific TnI phosphorylation on human cardiac tissue towards establishing this phosphorylation as a future therapeutic approach in human heart disease.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY / ABSTRACT Ependymomas are tumors that occur in the brain or spinal cord and are incurable in nearly half of all patients. Recent molecular classification has identified numerous molecular subgroups of ependymoma with supratentorial ependymoma ZFTA-RELA fusion positive (ZFTA-RELA) accounting for over 70% of all supratentorial ependymomas. ZFTA-RELA, a mainly pediatric brain tumor, has also been identified as one of the subgroups with the worst prognosis. These tumors arise from the oncogenic fusion between a central gene in the NF-κB pathway, RELA, and a gene with undescribed function, ZFTA. The resulting fusion protein, ZRfus, aberrantly recruits transcriptional and chromatin remodeling machinery to drive neoplastic transcriptional programs that includes constitutive activation of NF-κB signaling, activation of inflammatory gene expression programs, and stem cell programs. To date, chemotherapy has not become standard of care for any of the subtypes of ependymoma. All targeted therapies tested in ependymoma clinical trials have failed. Therefore, there is an urgent need to capitalize on the more recent molecular understanding of ZFTA-RELA to develop novel therapeutic paradigms that increase survival outcomes for these patients. In the search for co-regulatory proteins as candidate tumor dependencies and targets, we identified the chromatin remodeling complex, FACT (FAcilitates Chromatin Transcription), as a ZRfus interacting protein. Moreover, FACT is elevated in ZFTA-RELA compared to normal brain tissue and other ependymoma disease subtypes. Project goal: To thoroughly investigate FACT as a driver of ZFTA-RELA and to reveal it as a promising therapeutic target for this devastating disease. FACT, a heterodimer of SPT16 and SSRP1, mainly serves to reorganize nucleosomes to facilitate RNA polymerase II-mediated transcription. In tumors, we and others have shown that FACT is essential for maintaining an undifferentiated stem-like state necessary for tumor growth. This is relevant for ZFTA-RELA tumors as they are characterized as having undifferentiated transcriptional profiles. Project hypothesis: FACT regulates ZRfus oncogenic and inflammatory genes to maintain an undifferentiated cell state. Compromising FACT function will lead to reduced tumor growth, modulate tumor inflammation, and improve survival in orthotopic murine tumor models. Aim 1: Determine if FACT is essential for sustaining transcription of ZRfus targets (including oncogenes and inflammatory genes) and stem cell identity genes that may be important for tumorigenicity. Impact: These studies will reveal how FACT regulates ZRfus transcription and tumor cell identity. Aim 2: Evaluate the impact of genetic and pharmacological disruption of FACT on tumor progression, immune landscape, overall survival, and normal neurogenesis. Impact: These studies will reveal preclinical insight into FACT as a therapeutic target, and the efficacy of our candidate small molecule anti-neoplastic as rationale therapy to inform future clinical trial design. Overall, successful completion of our studies will reveal new therapeutic options for ZFTA-RELA ependymoma that can rapidly be moved into clinical trials.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Nonsense mutations pose a serious challenge to fitness and survival of cells and organisms. To suppress mRNAs carrying such nonsense mutations, all eukaryotes possess a conserved mRNA surveillance pathway called Nonsense-Mediated mRNA Decay (NMD). NMD is also an essential post-transcriptional regulator of normal mRNAs that shapes processes such as stem cell maintenance, neurogenesis, germ cell development and anti-viral response. In all eukaryotes, NMD is governed by three UPF proteins, UPF1, UPF2 and UPF3. In multicellular organisms, NMD is also regulated by a conserved multi-protein exon junction complex (EJC), which binds upstream of mRNA exon-exon junctions. During translation, if at least one EJC remains present downstream of a terminating ribosome, it can signal premature termination and trigger NMD. Understanding NMD mechanism and its regulation by EJC is crucial for betterment of human health as mutations in EJC and NMD proteins cause developmental defects, intellectual disability and mental retardation. The overarching goal of this research program is to understand how the remarkable variation in composition and function of EJC/UPF machinery regulates NMD to dictate cellular function and fate in animal cells. To achieve this goal, we are using a combination of genetic, genomic, molecular, biochemical and cellular approaches in cultured human cells and in zebrafish embryos to pursue four main directions. (1) We will identify the mechanism of a switch in EJC composition that we recently discovered and define the role of distinct EJC compositions in gene expression. (2) Our recent discovery that mammalian UPF3 paralogs and their interaction with EJC are non-essential for NMD challenges a decades old model of EJC-dependent NMD in eukaryotes. We will apply new genomic technologies that probe in vivo ribosome function to identify the role of UPF3 and other UPF proteins in premature termination complex assembly and activity on hundreds of human mRNAs. We will also identify the factors and features that govern signaling between the termination complex and the EJC. (3) We and others have previously shown that EJCs are often detected at unexpected locations on RNAs. By exploiting a new step in EJC recycling that we have uncovered, we will define the assembly mechanisms and functions of EJCs at such unexpected sites. (4) We have developed zebrafish mutants that lack one of the EJC or its NMD adapter proteins, which will be used to identify the genetic and cellular processes controlled by these factors during motor neuron and muscle development. Overall, our work will advance the knowledge of NMD mechanisms and how they regulate post- transcriptional gene regulation to control cellular function and organismal development. This progress will also elevate our ability to target NMD for therapeutics.

NIH Research Projects · FY 2026 · 2023-03

Sepsis, the injurious systemic response to infection, is a major cause of death in both industrialized and developing societies. With advances in supportive care, sepsis mortality has improved, leading to increased recognition of the chronic consequences of this critical illness. Nearly 50% of sepsis survivors experience long- term cognitive impairment, akin to accelerated dementia. Despite the substantial societal burden of septic cognitive impairment, no effective therapies are known to prevent or treat this condition. A multidisciplinary collaboration between the laboratories of Drs. Eric Schmidt (an expert in sepsis and vascular glycobiology) and Paco Herson (an expert in brain injury) recently identified a novel mechanism underlying the development of septic cognitive impairment. As detailed in manuscripts published in 2019 in the Journal of Clinical Investigation and the Proceedings of the National Academy of Sciences, we observed that heparan sulfate (HS) fragments, shed into the circulation during septic degradation of the endothelial glycocalyx, specifically penetrate into only one tissue: the hippocampus, a compartment of the brain responsible for spatial memory formation. Hippocampal-penetrating HS fragments sequester a key growth factor necessary for learning, leading to persistent cognitive impairment in both septic animals and humans. The striking hippocampal specificity of HS extravasation during sepsis prompted our laboratory to pursue additional preliminary experiments using a murine lipopolysaccharide (LPS) model of sepsis. LPS-induced hippocampal blood-brain barrier (BBB) hyperpermeability was specific to HS, as similarly-sized dextrans were unable to penetrate the hippocampus. Heparin affinity chromatography and proteomic analyses identified that LPS selectively induces several HS-binding proteins within the hippocampus, including an endothelial protein previously implicated in paracellular BBB permeability (Prion related protein, Prp). This selective induction of hippocampal HS-binding proteins mirrors the hippocampal specificity of HS extravasation during sepsis. Based upon these preliminary data, we hypothesize that sepsis induces expression of HS-binding proteins (such as Prp) released into the blood stream. These proteins facilitate the selective transport of circulating HS into the hippocampus, leading to cognitive impairment in sepsis survivors. Pursuit of this hypothesis will require mechanistic interrogation of sepsis-induced HS-binding proteins as putative mediators of HS uptake by the hippocampal BBB. To achieve this goal requires a multi-disciplinary team to use cutting-edge molecular tools to assess HS transport in animal models and clinical studies to correlate to the human patient population. We will specifically identify 1) the mechanism of HS accumulation into the hippocampus, 2) the role of circulating prion-related protein in neurocognitive dysfunction following experimental sepsis and 3) correlate circulating HS and Prp with cognitive outcome in human sepsis patients

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY / ABSTRACT Central nervous system (CNS) injuries such as stroke and spinal cord injury (SCI) are major contributors to the global burden of disability. Attenuating CNS damage represents the core of research and neurorehabilitation strategies to enhance recovery. Yet, mounting evidence of peripheral nervous system (PNS) alterations after CNS injury provides an untapped area for therapeutic investigation. The PNS links CNS motor output with skeletal muscle function, where motor unit recruitment and firing rate modulate control. A motor unit is comprised of one motoneuron and all myofibers it innervates. For precision of motor control, healthy myofibers receive one motoneuronal axon via a single neuromuscular junction (NMJ). Studies suggest profound motor unit loss in paretic muscle after stroke and SCI, though exact mechanisms are undefined. Moreover, the applicant recently identified striking NMJ remodeling after stroke, including aberrant polyaxonal innervation (PAI), where NMJs receive more than one axonal input. Pilot data in SCI demonstrate motor unit losses similar to stroke, but impacts of SCI at the NMJ remain unexplored. Taken together, this project will interrogate maladaptive PNS remodeling in the context of CNS injury disability. This work will test the therapeutic potential of targeting paretic NMJs with brain-derived neurotrophic factor (BDNF), a known mediator of motor neuron viability and NMJ plasticity. In murine models of stroke and SCI, the applicant will longitudinally study motor behavior, motor unit electro- physiology, and muscle contractility; assess histopathology of motoneuron pools and NMJs; employ molecular biology techniques to define mechanisms of PAI; and validate a novel gene therapy approach. Aim 1 will test the hypothesis that SCI induces motor unit dysfunction and NMJ remodeling, similar to stroke. Aim 2 will define pathophysiological mechanisms of stroke-induced motor unit loss; some predict motoneuron degeneration is responsible, however we hypothesize re-expression of developmental mediators induces motor unit overlap, with PAI presenting electrophysiologically as spurious motor unit loss. Aim 3 will test the hypothesis that post- stroke reduction in BDNF signaling drives PAI, while normalization of BDNF via adeno-associated viral delivery restores NMJ form and motor function. Mentored training in translational neuromuscular physiology from the Sponsor (a neuromuscular specialist with extensive preclinical/clinical experience in neuromuscular health and disease) is complemented by a gene therapy specialist in CNS/PNS diseases as Co-Sponsor, and supported by two key Collaborators (an SCI physician-scientist, and a neuroscientist with BDNF signaling expertise). This mentorship team dovetails with the excellent resources and environment at The Ohio State University to facilitate growth in new areas of investigation and prepare the applicant for independence. Using clinically-relevant approaches to interrogate peripheral mechanisms of motor dysfunction after CNS injury, this project will expand the fundamental understanding of stroke and SCI disability, inform future therapeutics targeting peripheral alterations, and offer critical training opportunities for career development in academic neurological research.

NIH Research Projects · FY 2026 · 2023-03

Abstract Henipavirus (HNV) genus, named after the first two identified members, Hendra virus (HeV) and Nipah virus (NiV), is a group of expanding zoonotic viruses that have caused repeated outbreaks with case fatality rate reaching 75%. The exceptional broad species tropism and various transmission routes make HNV a risk of potential future pandemics. Both HeV and NiV have been categorized as Biological Safety Level-4 (BSL-4) transboundary agents, and NIAID Category C Priority Pathogens. There are six emerging HNVs reported in recent years, showing limited antigenic cross-reactivity with NiV and HeV. Glycoproteins F and G are the two only spikes on the HNV surface, which coordinate the viral entry process via G glycoprotein-mediated receptor attachment followed by the F glycoprotein-mediated membrane fusion between virus and host cell. Both G and F proteins are the targets of HNV-neutralizing antibodies. Vaccine and monoclonal antibody (mAb) countermeasure development focusing on these two HNV glycoproteins are now of critical and urgent importance to prepare for potential HNV spillover events. The ectodomain of HNV G glycoprotein is a homo- tetramer with each protomer composed of a globular head and a stalk region. The metastable tetrameric conformation has restricted the previous structural characterization of G protein to the monomeric head only, which is a major obstacle for a comprehensive understanding of the G protein-mediated mechanisms of HNV entry and antibody recognition. HNV F glycoprotein trimer transits from the prefusion to the post-fusion conformation during viral entry process. The metastable prefusion conformation can be preferably recognized by neutralizing antibodies, whereby a promising target for vaccine design and therapeutic development. While the glycoproteins of both HeV and NiV have been extensively studied, the glycoproteins of the emerging HNVs, which are genetically and antigenically distinct from the two prototypic HNVs, also extremely diverse among themselves, have not been well investigated. We have used rational designs to created soluble, stabilized, oligomeric glycoprotein ectodomain constructs from several emerging HNVs, including two phylogenically distant bat-borne HNVs, CedPV and AngV, to facilitate structural, functional and antigenicity characterization. We propose to use structural and structure-based functional analyses of these glycoproteins to assist delineation of the function of HNV glycoprotein-mediated entry. We will also define the antigenicity of these HNV glycoproteins with neutralizing antibodies, including conventional murine antibodies and camelid single-domain antibodies, to investigate their neutralization epitopes and mechanisms structurally and functionally. The combined antigenicity results will render a comprehensive definition of sites of vulnerability on both G and F glycoproteins of the emerging HNVs. Collectively, findings derived from this project will provide insights to HNV entry mechanism, as well as inform future work on designing envelope glycoprotein-based vaccine and immunotherapeutic against emerging HNVs.

NIH Research Projects · FY 2026 · 2023-02

The goal of this project is to determine the efficacy of computerized cognitive training for breast cancer survivors (BCS) suffering from cancer-related cognitive impairment (CRCI). For millions of cancer survivors, CRCI is a prevalent, severe, and persistent problem that negatively impacts work outcomes (work ability and productivity), health perception, and health-related quality of life. Evidence suggests that up to 75% of the more than 3.8 million BCS in the U.S. will experience cognitive changes that may persist for years after treatment ends. Unfortunately, the scientific basis for managing these cognitive changes in cancer survivors is extremely limited. Available evidence from pilot studies, including our own work, suggests that computerized cognitive training, which is based on the principles of neuroplasticity (ability of brain neurons to re-organize and form new neural networks), may be a viable treatment option. However, previous trials to date have been limited by lack of attention-controlled designs, small samples, and limited follow-up. Therefore, to overcome limitations of past studies and build on our pilot results, the purpose of this 2-group, double-masked, randomized controlled trial is to conduct the first full-scale efficacy trial to compare computerized cognitive training (BrainHQ) to computerized active attention control (Sudoku, crossword, word find, etc.) in BCS. Specific aims are to: (1) test the efficacy of computerized cognitive training on improving perceived cognitive function immediately post-intervention and over time, compared to active attention control; (2) test the efficacy of computerized cognitive training on cognitive performance over time compared to attention control; and (3) explore transfer effects on real-world, everyday outcomes including work-related outcomes and health-related quality of life over time compared to active attention control. This proposal has been peer-reviewed and endorsed by the NRG Oncology Research Base of the NCI Community Oncology Research Program (NCI NCORP) and the NCI Division of Cancer Prevention has approved NRG Oncology to conduct the trial at their affiliated sites. A total of 386 eligible BCS will be identified and consented through the NCI NCORP and NCI National Clinical Trials Network (NCTN) sites, composed of over 2,000 participating clinical oncology sites. Outcomes will be collected at four time points: baseline, prior to intervention (T1), immediately post-intervention (T2), 3 months (T3), and 6 months (T4) post-intervention. Data will be analyzed using linear mixed models for repeated measures. The current proposal responds directly to the NCI Notice of Special Interest to test interventions designed to address the adverse aging-related effects of cancer and cancer treatments, builds on our previous pilot studies while also making methodological improvements, and leverages access to all NCORP sites. Therefore, this will be the first full-scale study to test computerized cognitive training in cancer survivors with CRCI and provide empirical evidence for clinicians’ recommendations and survivors’ treatment selections for managing cognitive impairment.

NIH Research Projects · FY 2026 · 2023-02

Abstract Liver cancer is one of the most common types of cancer. More than 700,000 people are diagnosed with this cancer each year throughout the world. Liver cancer is responsible for more than 30,000 new cases and 12,000 deaths a year in the United States. Liver cancer has been found to be highly associated with ER stress/unfolded protein response (UPR)-induced chronic inflammation; however, the mechanisms remain unclear. Using a genetic approach, we have found that deletion of an ER protein, CNPY2, from macrophages prevents carcinogen-induced hepatocellular carcinoma, accompanied by reduced ER stress/UPR signals and tumor- associated macrophages in tumors. Furthermore, CNPY2 is required for pro-inflammatory cytokines, IL6 and TNFα released from Kupffer cells/liver macrophages. We also observed that CNPY2 plays a central role in regulating both ER stress/UPR and TLR4 signaling, two pathways known to promote cytokine production and differentiation of macrophages. Together these observations suggested that CNPY2 promotes liver oncogenesis through regulation of macrophages. In this proposal, I will aim to address several fundamental questions in the field of HCC: 1) what is the mechanism by which CNPY2 transcriptionally regulates production of IL6 and IL23 in macrophages. Are both the UPR and TLR4 pathways involved in this regulation? 2) what is the biochemical and structural basis for the roles of CNPY2 in promoting TLR4 signaling-dependent cytokine production in macrophages. Solving crystal structure of CNPY2 will help drug development against CNPY2. 3) The potential mechanism by which CNPY2 regulates differentiation, infiltration and function of tumor-associated macrophages in HCC. 4) What is the role of CNPY2 in tumor immunity? Does targeting CNPY2 improve anti-tumor immunity in HCC? Our traditional and conditional Cnpy2 KO mice are unique models for addressing these questions. Successful execution of this work will significantly advance the field. In the longer term, this study may lay a strong foundation for the development of a new class of therapeutics for cancer, based on the rational design of CNPY2 inhibitors against tumor-associated macrophages.

NIH Research Projects · FY 2026 · 2023-02

Project Summary Since its outbreak in China in December 2019, the global impact of SARS-CoV-2 infection has been extraordinary, with over 350 million cases and more than 5.5 million lives lost (WHO Coronavirus Dashboard, Jan 2022). Despite various public health measures, such as social distancing, handwashing, face masking, and vaccination, infections in the US and the world continue in waves driven by new variants of concern (WHO). While the pandemic landscape has been constantly shifting, epidemiologists and public health experts increasingly project that the virus could eventually become endemic, especially with emerging zoological reservoirs. While information is still emerging, the full picture of neurological consequences of COVID-19 to broad sensory functions remains unclear, especially in the context of new variants, vaccination, prior infection, and ongoing treatment. For example, smell loss has been a hallmark symptom of COVID-19 (>80%) and can present in isolation (the only symptom of COVID-19) or precede the occurrence of other symptoms. Epidemiology studies have shown that smell loss is the most predictive symptom for COVID-19, better in identifying COVID-19 patients than cough, fever, headache, or other typical symptoms used to screen for COVID-19 in workplaces, schools, and health care settings. But with the emergence of Delta and Omicron variants and vaccination, it is unclear whether olfaction function is still severely impacted as in the early stage of the pandemic. Similarly, taste and chemesthesis (trigeminal) losses have been implicated but not fully differentiated from the flavor loss caused by retronasal smell losses. Sudden onset of hearing loss and dizziness have also been self-reported among COVID-19 patients, but current evidence for these are still limited, despite being in the 3rd year of the pandemic. In this study, Aim 1 will apply multidisciplinary methodologies to extensively capture and quantify the full impact of COVID-19 on broad sensory functions (smell, taste, chemesthesis, hearing, balance/vestibular function), with endemic viral upper respiratory infection (URI: cold, flu, etc.) as a comparison group. These tests will allow us to extensively explore and differentiate the impact of COVID-19 on multiple sensory functions and their association with the disease profile, such as severity, dominant variant at the time of infection, vaccination status, prior infections, and treatment received, with potential to characterize common and distinct factors to that of URI. Aim 2 will broadly assess the longitudinal time course of sensory loss and recovery among COVID-19 patients, with the expectation that recovery can vary depending on the system affected and the disease profile. Aim 3 will specifically focus on COVID-19 “long haulers”, patients who have persistent symptoms (>90 days), to examine the different characteristics in sensory losses and recovery during the prolonged disease phase. The outcomes of this study could importantly expand our understanding of the characteristics of broad sensory losses and their recovery among COVID-19 patients.

NIH Research Projects · FY 2026 · 2023-02

Project Summary This K08 mentored clinical scientist research career development award is a five-year program designed to facilitate Dr. Marios Arvanitis’ (PI) development into an independent physician-investigator in vascular genetics. Atherosclerotic cardiovascular disease (ASCVD) is a major public health burden that accounts for over 600,000 deaths in the United States each year. ASCVD is highly heritable and genome-wide association studies have discovered many candidate genomic loci that increase the risk of the disease in the population, thereby providing a window to novel therapies. However, most genomic risk loci for ASCVD remain unexplored in terms of how they lead to disease risk. Previously published work by the PI has focused on the mechanistic interpretation of genomic risk loci for cardiovascular disease, including the development of a novel Bayesian method, called CAFEH, to prioritize the target tissue and genes in genomic loci. Our preliminary analyses of the genetic underpinnings of ASCVD reveal that endothelial cells are enriched for ASCVD heritability, and we have used those methods to prioritize a chromosome 4 locus that is predicted to affect ASCVD risk via altering the expression of the RE-1 silencing transcription factor (REST) gene in endothelial cells. This K08 project will explore the regulatory mechanisms via which the REST locus and gene influence the development of atherosclerosis. Aim 1 will employ CRISPR-Cas9 editing in human stem cells which will then be differentiated into endothelial cells to identify the causal variants and the upstream transcription factors that mediate the association in the 4q12 coronary disease GWAS locus. Aim 2 will define distal genes and pathways affected by REST in the endothelium and investigate their cellular consequences, starting with evaluating the role of REST in endothelial to mesenchymal transition. Aim 3 will use a tamoxifen inducible endothelial specific Rest knock-out mouse model to evaluate the in vivo effects of Rest in the endothelium and atherosclerosis. The success of this project is guaranteed by the support of a multidisciplinary mentoring team including a vascular biologist (Dr. Charles Lowenstein), a computational biologist (Dr. Alexis Battle), and a functional genetics expert (Dr. Andrew McCallion), along with an advisory committee of experts in vascular biology, stem cell differentiation and atherosclerosis (Dr. Harry Dietz, Dr. Chulan Kwon and Dr. Thomas Quertermous). This award period will help the PI boost their genomics skills, acquire new wet lab skills and generate preliminary data to successfully compete for R01 funding in order to translate the genetic insights into novel mechanisms for ASCVD.

NIH Research Projects · FY 2025 · 2023-02

Project Summary/Abstract African Americans (AAs) with Type 2 diabetes (T2D) have worse glycemic control and a 50% higher mortality rate of diabetes compared to non-Hispanic Whites (NHWs). AAs with T2D have 3.2 times more hospital admissions for uncontrolled diabetes compared to NHWs. Multiple, intertwined factors at the individual, interpersonal, community, societal, and healthcare system levels contribute to lower adherence to diabetes self- management, greater difficulty achieving glycemic control, and higher rates of microvascular and macrovascular complications. The NIH Science of Behavior Change identifies interpersonal and social processes as one of the three key mechanisms for behavior change, providing greater support for this strategy. Family is a critical social context in which interdependence, collectivism, and extended family network is central to their way of life among AAs. Indeed, focusing on the individual-level demonstrated only limited improvements in glycemic control for AAs with T2D. Thus, effective multi-level interventions that promote adherence to diabetes self-management in this vulnerable population are sorely needed. Our proposed phase I/II randomized controlled trial will not only improve glycemic control for participants with T2D, but also engage family members in physical activity and healthy eating strategies. The specific aims are: 1. to examine the feasibility and acceptability of a family-dyad- focused diabetes intervention in AA adults with T2D and their designated family members; 2. to examine the preliminary efficacy of the family-dyad-focused diabetes self-management intervention compared to a waitlist control arm on: (1) glycemic control (hemoglobin A1c) and health-related quality of life (HRQOL) (primary outcomes); and (2) blood pressure control (secondary outcome) in participants with T2D; and 3. to explore the dyadic relationship (quality and support) and its association with a) changes in dyadic stress, physical activity and dietary intake, and b) health outcomes (glycemic control, HRQOL and blood pressure control) over time in participants with T2D. We will conduct a two-arm RCT. We will enroll 104 AAs with T2D and one family member of each patient (104 dyads), randomized 1:1 to intervention or wait list control arm (n=52/arm). All participants will undergo the standard usual care held at the pharmacy clinic. Patient-family-member dyads in the intervention arm will receive 1) 14 session over 20 weeks of family dyad-focused, in-person group sessions on diabetes self- management and family support; 2) family dyad-focused support component in each group session; and 3) individual family feedback telephone sessions. All participants will be assessed at baseline, post-intervention and six months after intervention. Our goals of the intervention are to encourage participants to (1) daily self- manage diabetes and stress; (2) establish a healthy eating pattern reducing overall calorie and carbohydrate intake; (3) engage in brisk walking of 150 or more minutes a week; and (4) use solution-focused problem-solving strategy and supportive family communication skills. Our approach will add to the scientific knowledge and identify interpersonal and social mechanisms in adherence to diabetes self-management among AAs.

NIH Research Projects · FY 2026 · 2023-02

Project summary: Traumatic brain injury (TBI) in childhood is the leading cause of pediatric emergency room visits, with over 800,000 children visiting the ER each year according to the CDC. Pediatric TBI can have lifelong consequences for behavioral health, increases rates of ADHD, drug and alcohol abuse, long-term cognitive and social deficits, depression and anxiety. This suggests that early childhood is a period of particular vulnerability to long-term, deleterious neurological outcomes after TBI. Despite the clear evidence of a significant public health problem, the proximal mechanisms leading up to those long term TBI-related outcomes are poorly understood. TBI induces robust neuroinflammation and brain-resident innate immune cells, such as microglia, regulate normal brain development, including synaptic patterning. The impact of TBI on microglia-synaptic interactions is poorly understood. We have demonstrated dramatic developmental biases in activation and sex differences in the profile of neuroimmune cells in the immature rat brain, both microglia and the less studied mast cells. Mast cells are abundant in the developing brain and sparse in adults, suggesting that mast cells could contribute uniquely to the neuroimmune milieu after pediatric TBI. Mast cells are ‘first responders’ to immune insults and coordinate subsequent immune cell (microglia and astrocyte) activation as well as vascular permeability but their role in TBI has not been well studied. In our project, we will use lateral fluid percussion injury on juvenile rats to model pediatric TBI, which our preliminary data suggest elicits robust mast cell activation in the hippocampus, acute gliosis, and long-term, sex-dependent shifts in social behavior and stress hormones. Because so little is known about the unique pediatric response to injury, we will compare the pediatric versus adult injury response of male and female rats via RNAseq and Nanostring profiling of isolated immune cells in a comprehensive time course study (Aim 1). To test for a contribution of mast cells to pediatric TBI, we will use an acute mast cell inhibition using an FDA-approved pharmacological agent and comprehensively profile neuroinflammatory responses, alterations in blood brain barrier (BBB) permeability to narrow in on a potential mechanism through which mast cells are acting after TBI, and correlate BBB changes with social and stress-related behavior outcomes (Aim 2). To determine whether microglia are important for long-term neurodevelopmental programming of behavioral outcomes and sculpting neural circuits after pediatric TBI, we will perform microglia depletion/forced turnover experiments post-TBI (Aim 3). We will compare sexes in all studies, and we predict that males more robust basal neuroimmune tone in the developing brain may render them more vulnerable to TBI-related outcomes. Our studies have the potential to improving long-term outcomes following pediatric traumatic brain injury and uncover potential new therapeutic options targeted to the unique neuroimmune environment of the developing brain.

NIH Research Projects · FY 2026 · 2023-02

For decades, the tobacco industry has manipulated moist snuff (i.e., dip) to become as appealing and addictive as possible. These manipulations have resulted in moist snuff accounting for 90% of the smokeless tobacco (SLT) market share in the United States and have slowed progress in reducing the prevalence of SLT use. The public health burden of the tobacco industry’s actions is especially high among males living in Appalachian regions of the United States who continue to use SLT at high rates, leading to increased cardiovascular disease, cancer, and poor oral health. The goal of this research is to identify local, state, and/or federal regulations for moist snuff that reduce its addiction potential. This research will evaluate two features of moist snuff that are key targets of manipulation from the tobacco industry and drivers of its addiction potential: length of tobacco cut (long vs. fine) and nicotine form (low vs. high levels of nicotine in the free-base form). Finer tobacco cuts and higher levels of free-base nicotine (FBN) result in faster, greater nicotine delivery. Building on the PI’s areas of expertise and providing crucially-needed training, evaluations of tobacco cut and nicotine form will be done epidemiologically, using analytical chemistry, and using experimental human laboratory methods. For the epidemiological evaluation, we will complete a secondary data analysis of recent population-based cohorts to evaluate associations between level of SLT dependence and product characteristics (i.e., cut, flavor, brand) of moist snuff used by adolescents and adults in Appalachian Ohio. For the analytical chemistry evaluation, we will quantitate the levels of total nicotine and FBN present in moist snuff products used by adolescent and adult participants in our Appalachian Ohio cohorts; these characteristics of products will be compared according to level of SLT dependence of the user as well as other product features (e.g., tobacco cut and flavor). For the human laboratory evaluation, we will use a 2x2 within-subjects cross-over design to test how moist snuff use (amount and duration), nicotine delivery, and appeal differ according to variations in cut (long vs. fine) and level of FBN (low vs. high). We will also evaluate whether the association between style of moist snuff and outcomes varies according to level of SLT dependence. Participants will be adults who live in Appalachian Ohio and are daily moist snuff users. Altogether, this research will provide a foundation for understanding the characteristics of moist snuff used in Appalachia—where the burden of SLT use remains stubbornly high—and the effects that regulations on tobacco cut and nicotine form could have on improving health. The conduct of this research and implementation of the proposed training plan will also provide the PI, a promising scientist with strong foundations in epidemiology and biostatistics, Appalachian health research, and other areas of tobacco regulatory science with needed training in addiction research, tobacco product evaluation, and human laboratory experimental designs.

NIH Research Projects · FY 2024 · 2023-01

Axons and their associated glia (Schwann cells and oligodendrocytes) form the largest part of the neuronal network. Axons are challening to maintain energetically and are vulnerable to a wide spectrum of noxious stimuli. Dysfunction of axons and pathological axon degeneration (pAxD) have emerged as a major pathophysiological driver in many neurodegenerative diseases. Consequently, a central therapeutic focus is to develop approaches tailored to protect axons. A prerequisite for such therapies is a better understanding of the autonomous and non- cell autonomous molecular mechanisms that regulate the processes leading to pAxD. Physical disconnection of the axon from the neuronal cell body is a widely-used experimental platform that has dramatically improved our understanding of these processes over the last two decades. Primarily studied in the peripheral nervous system of vertebrates, this paradigm triggers early injury responses in Schwann cells followed by rapid and stereotyped disintegration of axons (Wallerian degeneration). It is now known that axon disintegration is evoked by a conserved auto-destruction program that exhausts axonal ATP content through rapid depletion of the metabolic cofactor NAD+ in disconnected axons. Importantly, recent studies indicate an instructive role of axonal bioenergetics for the survival of injured axons. Given that neurodegenerative diseases are broadly associated with axonal bioenergetic defects, these findings suggest that the decline of axonal bioenergetics occupies a central position in the pathway leading to pAxD. In support of this, we recently made the exciting discovery that Schwann cells convert their energy metabolism early upon axon injury to antagonize the structural breakdown of injured axons, likely through the increased supply of glycolytic end-products (axon-glia metabolic coupling). Furthermore, we found that the manipulation of the metabolic injury adaptation in Schwann cells accelerates or delays the degeneration of perturbed axons in acute and subacute pAxD models. For the first time, this demonstrates a non-cell-autonomous energetic mechanism that controls the fate of injured axons. The first aim of this proposal attempts to determine if the suggested metabolic coupling mechanism counteracts the energetic decline of injured axons through the enhanced supply of glial manocarboxylates that support axonal ATP production. The next objective extends the identification of the key components of the metabolic coupling pathway critical for the support of injured axons with an emphasis on axonal mitochondria. The final goal intends to examine as to how manipulation of metabolic coupling influences pAxD in an iatrogenic disease model of subacute axon pertubation. Collectively, this work has the potential to introduce a paradigm shift away from neuron-centric views of axon protection. The proposed efforts may open the door for the future development of novel therapeutic approaches taking into account the relationship between axonal and glial bioenergetics to combat pAxD in neurodegenerative disorders. .

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY/ABSTRACT Understanding the causes and mechanisms underlying circadian rhythm disruptions that are associated with fatigue during cancer treatment remains unclear. This current deficiency means that successful cancer treat- ment falls short of its potential and prior quality-of-life remains elusive for patients. Our long-term goal is to im- prove debilitating behavioral sequelae in cancer patients, thus improving quality-of-life, other comorbidities, and mortality. Thus, the overall objective here is to establish the potential role of circadian disruption as a fun- damental pathway by which chemotherapy promotes cancer-associated fatigue. Indeed, robust circadian rhyth- micity of virtually all physiology is extremely well-conserved; desynchrony of these rhythms leads to negative health and behavioral consequences. The central hypothesis is that chemotherapy-induced inflammation inhib- its SCN function leading to fatigue. The rationale for this work is that circadian circuitry disruption is an under- studied, relevant pathway in psycho-oncology research that could elucidate mechanisms and new, rhythm-fo- cused interventions. Three specific aims are proposed to test the central hypothesis using our novel breast cancer “survivor” mouse model. Aim 1 will determine the ability of the master clock to entrain after chemother- apy. Behavioral SCN rhythm adaptations to environmental challenges will be assessed. Aim 2 will identify the role of central inflammation in master clock disruptions after chemotherapy. The role of chemotherapy-induced neuroinflammation on SCN molecular and behavioral rhythms will be quantified. The potential resolution of fa- tigue will also be assessed. Aim 3 will determine the role of circadian disruption in chemotherapy-induced fa- tigue. Genetic and pharmacological SCN timing manipulations will precede a battery of behavioral assess- ments of the physical, motivation, and cognitive components of fatigue. In vivo and ex vivo circadian timing ap- proaches combined with systems-, cellular-, and molecular-level analyses will pinpoint the effects of two regi- mens of chemotherapy on master oscillator circadian circuitry relevant to cancer-related behavioral comorbidi- ties. The proposed research is conceptually innovative because using circadian approaches is new to psycho- oncology. It is also technically innovative by way of the superior translational model and the circadian genetic and pharmacological techniques planned. This research will result in essential new knowledge about how com- mon cancer treatments affect the pacemaker, which is crucial to extensive downstream physiology and behav- ior (i.e., beyond fatigue). Results will provide much needed evidence to make circadian-based approaches standard in clinical practice, as well as inform the design of novel circadian-directed pharmacological and non- pharmacological interventions. This research is applicable to other cancers and in non-oncological populations treated with chemotherapy (e.g., stem cell transplant, lupus).

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Neurofibromatosis type 2 (NF2) is a profoundly devastating tumor predisposing syndrome characterized by multiple nervous system tumors that cause substantial debility for patients. The hallmark of NF2 is bilateral vestibular schwannomas (VS) which cause hearing loss, imbalance, brainstem compression and even death. Both NF2-associated and sporadic VS can develop aggressive cystic degeneration which result in rapid neurologic decline. There are no FDA-approved pharmacotherapies against cystic VS and surgery is the only treatment. However, surgical outcomes are highly variable, and surgery carries considerable morbidities including deafness, facial paralysis, brain injury and incomplete resection, due to significant peri-tumoral adhesions and reliance on imprecise visual cues during tumor dissection. Efforts are needed to identify biomarkers to classify aggressive VS and develop intraoperative tools to detect tumor cells, visualize the interface between tumor and cochlear/facial cranial nerves and improve the accuracy of surgical resection. This K08 career development award is designed to launch the principal investigator’s career as an independent surgeon scientist, whose goal is to understand mechanisms that underlie VS growth and cystic degeneration, identify molecular biomarkers to classify disease, and leverage nanotechnology tools to improve the precision of tumor surgery. The PI’s mentor (Krystof Bankiewicz) is a leader in delivering therapeutics to the central nervous system and translational research. The PI’s co-mentor (Long-Sheng Chang) is an expert in studying the NF2 tumor suppressor gene function in VS. Additional mentorship is provided by a research advisory committee of experts in nanoparticle delivery (Yizhou Dong), matrix metalloprotease and extracellular matrix biology (Jennifer Leight), and skull base surgery (Oliver Adunka), along with didactics and workshops at the Ohio State University and nationally. Previously, the PI utilized primary human VS cultures to develop nanoparticles that target VS and identified a matrix metalloproteinase whose activity was associated with poor surgical outcomes in VS. The proposed study will: 1) systematically identify proteases dysregulated in VS through spatial transcriptomic profiling; 2) engineer a library of nanomaterials to rapidly quantify VS protease activity and establish an activity-based tumor-classifying biomarker; 3) develop protease-sensing, tumor-targeted nanoparticles. Results from this work will elucidate mechanisms that regulate VS behavior and establish tools to enhance the accuracy of surgical resection, which could ultimately reduce treatment-associated morbidity, preserve cranial nerve function and improve outcomes for patients with this devastating disease. Furthermore, the principal investigator will obtain the training needed to transition into an independent physician scientist focusing on using nanotechnology to translate biological discoveries into new diagnostics and therapeutics for VS and NF2.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Among patients with diabetes, cardiovascular diseases (CVDs) are the primary cause of their mortality. Reducing CVD risk is a critical clinical goal for treating diabetic patients. Diabetes exacerbates atherosclerosis development and progression, which is the major cause of many CVD, including heart attacks, strokes, and peripheral vascular disease. Vascular smooth muscle cell (VSMC) dysfunction contributes to the pathogenesis of atherosclerosis throughout all the stages. The genetic relationship between diabetes and CVD provides the promise for the prevention and treatment of both disorders. Recent genetic studies have demonstrated that the specific variants at the coiled-coil domain containing 92 (CCDC92) locus are associated with both type 2 diabetes (T2D) and coronary heart disease (CHD). The biological function and detailed mechanisms by which CCDC92 regulates these diseases, a necessary step towards the ultimate goal of targeting CCDC92, remain unclear. Our preliminary data demonstrated that Ccdc92 knockout inhibits high-fat diet-induced insulin resistance and atherosclerosis in mice. We further present extensive preliminary studies showing that CCDC92 induces proatherogenic phenotypes, contributing to atherosclerosis pathogenesis. Here we hypothesize that VSMC CCDC92 promotes atherosclerosis development and progression by regulating the lysosomal pathway. By taking advantage of our unique animal models combined with molecular, cellular, histological approaches, we will define the role of CCDC92 in proatherogenic phenotypes in VSMCs in vitro (Aim 1); Determine the role of CCDC92 in atherosclerosis under diabetic and euglycemic conditions in vivo (Aim 2). Successful completion of the proposed study would provide a deep understanding of how CCDC92 elicits atherosclerosis and will likely set a profound foundation to define CCDC92 as a novel therapeutic target to treat atherosclerosis and diabetes-associated CVD.

NIH Research Projects · FY 2026 · 2023-01

Project Summary/Abstract There is an increasing need to improve the characterization of lipids and saccharides for clinical and biomedical research purposes. NMR is the method of choice for obtaining detailed structural information about saccharides. However, NMR typically requires milligram (micromole) quantities of analyte; this often exceeds biologically relevant levels. For lipids, mass spectrometry (MS) provides an efficient avenue for rapid profiling, but quantitative analysis is challenged by difficulty in isolating species of interest due to wide structural diversity. A new MS approach is proposed that fundamentally addresses challenges in quantitative and qualitative MS by utilizing online accelerated droplet chemistry. Since ion suppression effects in electrospray ionization (ESI) MS occur during droplet formation, our method is designed to tackle this intellectual challenge exactly at the point of droplet formation – not before by adding reagents in solution, and not after by performing gas-phase reactions. This strategy simplifies instrumentation requirements and allows effective coupling to liquid chromatography (LC). Selected droplet-based reactions improve signal-to-noise ratios to enable femtomole sensitivity using <1 µL sample volume. Gas-phase ion intensities generated by our platform reflect the corresponding analyte concentration in solution. Importantly, selected droplet-based reactions allow isomers of lipids and saccharides to be differentiated. We propose to couple online droplet reactions with LC to enable high throughput quantification of lipids and saccharides in complex mixtures. The specific research aims are: Aim 1: To develop a functional contained-electrospray platform for coupling accelerated droplet chemistry on LC-MS for saccharide analysis. A novel contained-ESI source is proposed to couple droplet chemistry with LC-MS. Our method will enable LC mobile phase and ESI spray solvent to be independently optimized. This orthogonal feature is expected to allow effective separation of isomeric saccharides (linkage, anomeric, and position isomers). Selected droplet reactions will improve detectability of saccharides and provide a second layer of identification for isomers that co-elute. The LC-contained-ESI-MS/MS platform will be validated via high throughput combinatorial studies. Aim 2: To develop a plasma-droplet fusing contained-electrospray source for coupling LC-MS for lipid analysis. We propose to include etched silica capillaries on our LC-contained-ESI-MS/MS platform for accurate quantification of all types of lipids, including triglycerides. The device is expected to enable instantaneous determination of degree of unsaturation, C=C bond position, and bond orientation (cis/trans). The tandem development of quantitative analytical methods for lipids and saccharides will result in concomitant creation of versatile platforms for applications in diseases diagnosis and high throughput analysis of rare sugars to effectively guide synthetic method development. The proposed strategy will also be valuable in biomedical research using existing instruments without modification.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY / ABSTRACT Extracellular vesicles (EVs) are cell-derived membrane-bound structures released into extracellular spaces that navigate the bodily fluids and appear to support intercellular communication. Cancer cells release significantly higher numbers of EVs then their normal counterparts. Because the EV contents are derived from the cell of origin, molecular profiling of circulating EVs are being scrutinized as a non-invasive means for early cancer diagnosis, monitoring disease progression, and assessing response to treatment. However, EV-based clinical diagnostics have been limited by inadequate rigor and reproducibility of samples to specifically discriminate, isolate, and characterize normal and disease-associated EVs. Three analytical and conceptual challenges have prevented the identification of specific and reproducible EV- associated cancer biomarkers with clinical relevance: 1) no unbiased strategy has been developed to evaluate the limits of cancer detection using EVs; 2) the relative contribution of healthy tissues to the pool of circulating EVs is not known; and 3) a systematic analysis of the number and composition of circulating cancer-derived EVs during tumor development has not been performed. These complications have prompted us to design a general platform capable of evaluating the contribution of specific tissues to the pool of circulating EVs in otherwise health animals compared to animals undergoing cancer development. The system is based on an engineered EV marker developed from the tetraspanin protein CD63 (enCD63), which facilitates collection, visualization, and quantification of EVs released by specific cells and tissues. By restricting the expression of enCD63 to specific normal or neoplastic cells and tissues of genetically engineered mouse models, we will unambiguously examine the efficacy of EVs as biomarkers. We propose to use our innovative platform to: 1) perform an unbiased calibration correlating the number of EVs with their cells of origin (Aim 1); 2) define the relative contribution of healthy tissues to the pool of circulating EVs (Aim 2); and 3) to assess the specificity and sensitivity of EVs in cancer detection (Aim 3). This application focuses on Pancreatic Ductal Adenocarcinoma (PDAC), a deadly neoplastic disease with low survival rate that lacks specific and sensitive diagnostic tests. Completion of the proposed studies will contribute to the development of standardized procedures for the preparation, selection, and analysis of EV-based biomarkers.

NIH Research Projects · FY 2026 · 2022-12

Urogynecologic meshes are often implanted in women to treat stress urinary incontinence and pelvic organ prolapse, two most common pelvic floor disorders. While peri-operative glycemic control is the standard of care, women with diabetes experience ~5-fold higher risk of developing mesh-related complications such as mesh exposure into vaginal cavity and pelvic chronic pain. These complications significantly impair women’s life quality and increase health costs. Since diabetes affects 1 in 10 adult women in the US and the incidence continues to increase, defining the mechanism underlying the diabetes-associated risk of mesh complications is critical to improve patient care and decrease societal expenses. Studies have demonstrated that the beneficial effects of intensive glycemic control are markedly decreased if long episodes of hyperglycemia precede the treatment, referred to as “hyperglycemic memory” (HgM). To date, there has been no investigation into how this phenomenon impacts the outcomes of women receiving urogynecologic meshes. Our preliminary data in a rat model support that hyperglycemia is harmful to vaginal immune response to mesh in the long term and that HgM in bone marrow (BM) cells may impact mesh outcomes. Here we hypothesize that hyperglycemia leaves epigenetic marks in BM progenitor cells, which drives macrophage dysfunction at mesh-tissue interface, leading to an increased risk of mesh complications despite later glucose normalization. As epigenetic modification is reversible with long-term glycemic normalization, we further hypothesize that tight and long-term peri-operative glycemic control attenuates the negative impact of HgM, thereby improves mesh outcomes in women with diabetes. We have assembled an interdisciplinary team to test the hypotheses with the following specific aims: (1) Define the role of HgM in host response to mesh. In vitro experiments using hyperglycemia-impacted vs. normal BM cells will be performed to define impact of HgM on macrophage phenotype and function. In vivo experiments using BM transplantation in a diabetic rat model with mesh implanted via sacrocolpopexy will be performed to define the impact of HgM in BM cells vs. HgM in vaginal tissue on the long-term mesh outcomes at 90 days. (2) Define the effect of peri-operative glycemic control regimens on host response to mesh and long-term mesh outcomes. Three peri-operative glycemic control procedures – early and strict, immediately before surgery, and poor glycemic control will be implemented. Mesh outcomes at short-, medium-, and long-terms (i.e., 7-, 42-, and 90-days post-surgery) will be compared between groups. (3) Define the relationship between HgM and diabetes- associated risk of mesh complications via an in-depth analysis of host response in human mesh-tissue samples obtained from women with mesh complications. This study will provide insight into the mechanism of long-term implant complications in women with diabetes and inform future preventive strategies to improve health outcomes in this rapidly expanding demographic.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY The rising tide of antimicrobial resistance threatens catastrophic increases in mortality in the coming decades. Methicillin-resistant Staphylococcus aureus (MRSA) remains a leading pathogen. New antibacterial classes are urgently needed to ensure adequate therapeutic options for MRSA and other resistant bacteria. Novel Bacterial Type II Topoisomerase Inhibitors (NBTIs) derive their efficacy by targeting the clinically validated essential enzymes, DNA gyrase and topoisomerase IV (TopoIV). A novel binding mode avoids target-based cross-resistance to fluoroquinolones and establishes NBTIs as a new antibacterial class. A lead, gepotidacin, stands at the threshold of FDA approval, with several completed Phase 2 and ongoing Phase 3 clinical trials. Resistance to gepotidacin has been observed but is very poorly characterized. The transformative potential of the NBTIs will require a better understanding of mechanisms of action/resistance and new medicinal chemistry strategies to deliver highly efficacious successor NBTIs, the areas of focus in the present proposal. To date, we have synthesized >250 highly diverse NBTIs. Our anti-MRSA lead, 147, showed in vivo efficacy in two infection models and a favorable cardiovascular safety profile by rationally designed reductions of basicity and lipophilicity. We have generated NBTIs with improved dual-targeting of gyrase and TopoIV, reduced rates of spontaneous resistance, and greater antibacterial activity over gepotidacin against NBTI- resistant MRSA. In contrast to gepotidacin, several newly synthesized amide-containing NBTIs induced DNA double strand breaks which we will investigate as a new mechanism of action for the NBTI class. Critically, we also propose that studies with our existing and planned NBTIs, coupled with our demonstrated expertise in microbiology, biochemical pharmacology, computational chemistry, and structural biology, will effectively address major unanswered questions regarding the emergence of resistance to NBTIs and strategies to overcome this issue. Overall, our goal is to generate lead compounds as innovative chemical tools and/or clinical candidates for further development. Three integrated specific aims will be pursued by our interdisciplinary team to: 1) Synthesize structurally and mechanistically distinct NBTIs with druglike properties 2) Evaluate new NBTIs for antibacterial activity & identify/characterize key NBTI-resistant S. aureus mutants 3) Elucidate the mechanism(s) of action of and molecular resistance to new lead NBTIs Aim 1 serves as the innovation engine for the proposal. Aims 2 and 3 support Aim 1 through iterative cycles of rigorous assays to provide new lead compounds. New fundamental information concerning the origin, mechanism, and impact/circumvention of acquired resistance to NBTIs will advance this new class of antibacterials as a pathway to promote human health by addressing the crisis in antimicrobial resistance.

NIH Research Projects · FY 2025 · 2022-12

Project Summary Although adolescents (ages 11 to 17) make treatment gains (e.g., reduced internalizing or externalizing behaviors) in psychiatric residential treatment (RT), they experience significant difficulty adapting to the community and often do not sustain treatment gains long term. After RT, 70% of adolescents discharge to their family of origin. However, parents are not provided with the necessary support or behavior management skillset to bridge the gap between RT and home. A new federal mandate will soon require parent training, an evidence-based behavior management intervention, in the RT setting. Parents with adolescents admitted to RT are a difficult-to-reach population, and technology may increase access and uptake of parent training. Parenting Wisely (PW) is a web-based parent training with demonstrated efficacy in increasing effective parenting practices to reduce adolescent behavior problems. We previously found that PW was highly feasible for parents and the skills were perceived as useful. However, parents reported two unmet needs: (1) skill individualization to apply the PW skills and (2) enhanced community to reduce isolation. In collaboration with an advisory board (a partner in the proposed study), we augmented PW with clinician facilitated discussion groups (referred to as PWRT). The discussion groups in PWRT supports program completion and parent engagement, provides a venue to discuss individualizing PW strategies, reduces isolation, and support parents by engaging in conversation about parenting in the RT context. The proposed study aims to: (1) establish feasibility and acceptability of PWRT, (2) evaluate whether PWRT engages target mechanisms (parental self- efficacy, parenting behaviors, social support, family function), and (3) determine the effects of PWRT on adolescent outcomes (internalizing and externalizing behaviors, placement restrictiveness). Sixty parents (30 per condition) will be randomly assigned to PWRT or treatment-as-usual (TAU). Each week for six weeks, parents will complete two PW modules (20 minutes each) and attend one discussion group via Zoom (90- minutes). PWRT will be initiated towards the end of the RT admission and continue post-discharge to bridge the transition from RT to the community. Adolescents (n=60) will not receive intervention; however, we will evaluate the feasibility of adolescent data collection for future studies. Data from parents and adolescents will be collected at baseline, 6-weeks, and 6-months post-baseline to allow for a robust understanding of the longer-term effects of PWRT on treatment gain maintenance. Consistent with PAR-21-211, our team of researchers and community partners is collecting the requisite data for a larger-scale effectiveness trial by testing the feasibility, acceptability, and preliminary effects of PWRT vs. TAU. PWRT is among the first web- based parent training augmented with supportive elements designed to engage parents with adolescents in RT. By providing parents with tailored education and support in PWRT, parents will be equipped with the behavior management skillset to provide structure in the home and ultimately maintain RT treatment gains.