Ohio State University

NIH Research Projects · FY 2025 · 2024-06

Project Summary/Abstract: Heart failure (HF) is a leading cause of morbidity and mortality worldwide, with projected numbers continually rising, mandating a need for novel therapeutic approaches. A common feature in the development of HF is hypertrophic growth of cardiac myocytes and associated remodeling of the size, dimensions, and function of the heart. Pathologic hypertrophy initially occurs as an adaptive response, leading to increased width of individual myocytes and causing concentric growth characterized by thickened heart walls, reduced wall strain, and maintained function. Left unchecked, this hypertrophic growth becomes maladaptive and reorients to growth along myocyte length, causing relative wall thinning, heart dilation, and declining function leading to HF. We currently have a poor understanding of the mechanisms which govern this transition, yet, limited observations where adaptive growth is preserved shows resistance to HF development. Therefore, this proposal seeks to identify the fundamental mechanisms underlying adaptive and maladaptive hypertrophic growth and investigate targeted interventions to maintain and/or restore the adaptive state for HF prevention. This proposal will address the critical distinction that not all pathologic hypertrophy is adverse and that preserving the adaptive, concentric state is therapeutically advantageous in response to chronic stress. Preliminary data has implicated a role for the phospho-regulation of the transcription factor STAT3 in mediating this transition. In particular, phosphorylation of the serine residue 727 on STAT3 was revealed as a critical target with dramatic influence over concentric/eccentric growth. Therefore, our central hypothesis is that STAT3 Ser727 phosphorylation is directly responsible for the induction of gene programs which drive adaptive versus maladaptive hypertrophy and represents a therapeutic target in HF treatment. The approach will be to: 1) Determine the molecular mechanism linking STAT3 Ser727 phospho-regulation to hypertrophic orientation. 2) Define novel gene targets and pathways which tune cardiac myocyte growth and hypertrophy. Specifically, this approach will address altered STAT3 transcriptional activity dependent on Ser727 phosphorylation through ChIP-seq and RNA-seq to identify gene programs which enact concentric/eccentric states. 3) Lastly, we will test novel therapeutic strategies to support adaptive cardiac remodeling during pathologic hypertrophy in vivo to assess effectiveness in HF prevention. Overall, we anticipate that these data will expand our understanding of HF remodeling, delineate the nature of adaptive, concentric hypertrophy, and reveal novel therapeutic opportunity in HF. Furthermore, characterization of STAT3 phospho-regulation and transcriptional activity will provide significant pathophysiologic insight to numerous other disease states such as cancer, fibrosis, inflammation, and immune signaling where STAT3 activity has been implicated.

NIH Research Projects · FY 2025 · 2024-05

Among more than 180 types of RNA modification, ribose 2’-O methylation (or called Nm, where N stands for any nucleotide) is the second most abundant. In 2’-O methylation, a methyl group is added to the 2' hydroxyl of the ribose moiety of a nucleoside. There are two types of Nm, the cap Nm which is the 2’-O methylation on the 5’ mRNA cap structure, and the internal Nm which occurs in the body of an RNA molecule. Cap Nm is extensively studied and its function is known. However, the enzymatic activities and biological functions of internal Nm are currently unknown. Viral RNA modification is a key step in the SARS-CoV-2 replication cycle. In addition to the 5’ cap guanine N-7 methylation and ribose 2’-O methylation (cap Nm), SARS-CoV-2 RNA is heavily epigenetically modified. We recently developed a high throughput Nm-Mut-seq technique allowing for mapping internal Nm sites at a single base resolution. Using this technique, we found for the first time that SARS-CoV-2 RNA isolated from well-differentiated primary human bronchial epithelial (HBE) cultures and Vero-E6 cells contains 24 and 11 internal Nm sites, respectively. Thus, the goal of this R21 project is to identify enzymes that install internal Nm sites in SARS-CoV-2 RNA and to explore the biological functions of internal Nm sites in SARS-CoV-2 RNA. In Aim 1, we will determine if any known host RNA 2’-O methyltransferase (MTase) or viral nsp16 protein (the only known viral RNA cap 2’-O MTase) installs Nm on SARS-CoV-2 RNA. As a parallel strategy, we will also use RNA affinity chromatography, mass spectrometry, and bioinformatics to identify this unknown 2’-O MTase. In Aim 2, we will test the hypothesis that internal Nm in SARS-CoV-2 RNA increases RNA stability, enhances mRNA translation, and prevents viral RNA from being recognized by host innate immunity. The Nm sites in viral RNA will be mutated and recombinant SARS-CoV-2 lacking internal Nm will be recovered using a SARS-CoV-2 reverse genetics system. The effects of internal Nm on viral RNA stability, protein translation, and innate immune response will be determined. This project will have an impact because it will not only fill a major gap in our understanding of the biological functions of internal Nm in SARS-CoV-2 RNA but will also facilitate the development of therapeutic agents for SARS-CoV-2 by targeting internal Nm.

NIH Research Projects · FY 2025 · 2024-05

Summary Bordetella pertussis (Bp), a gram-negative bacterium is the causative agent of whooping cough or pertussis, an acute disease primarily in infants and young children. Despite high vaccination coverage, pertussis is resurging in many countries including the USA. Bp infection in vaccinated individuals cause mild symptoms or are asymptomatic. This results in severe underreporting of global pertussis incidence. Current acellular pertussis vaccines (aPV) elicit suboptimal and short-lived immunity and fail to prevent the colonization of the nasal cavity. These infected individuals serve as a reservoir for bacterial transmission. Since Bp is restricted to humans as hosts and has no environmental or animal reservoir, it is critical to utilize model systems that resemble the environment of the human respiratory tract. Traditionally, the focus of Bp research has been on studying its interactions and pathogenesis in the context of lower respiratory tract and by utilizing animal models. The mechanisms utilized by Bp to survive and establish persistent infection in the human nasal cavity are poorly understood. We hypothesize that attachment to nasal epithelium followed by biofilm formation are key determinants of long-term infection of Bp in the human nasopharynx. In this proposal, we will use primary well- differentiated human nasal epithelial cultures (HNEC) grown at the air-liquid interface. These primary HNECs produce mucus, are ciliated and mimic the human nasal environment. In Specific Aim 1, we will determine (i) how Bp attaches, establishes colonizes and forms biofilms on HNEC; (ii) the role of known virulence factors in facilitating HNEC infection and (iii) how Bp infection alters the cellular and morphological characteristics of HNEC. Infection often elicits a dynamic cascade of events resulting in the adaptation of both the host and the pathogen. The crosstalk between Bp and the nasal cells following infection is yet to be investigated. We hypothesize that infection of nasal cavity by Bp triggers transcriptional changes in both the bacterial and host cells resulting in shaping of the infection process. In Specific Aim 2, we will use high throughput dual RNA sequencing technology to analyze the changes in the transcriptome of both Bp and HNEC. With the proposed research, we will gain an advanced understanding of host-pathogen interactions and identify the dynamic changes occurring in both the host and bacterial transcriptome during infection.

NIH Research Projects · FY 2025 · 2024-05

Project Abstract Treatment of biofilm-associated infections, such as those caused by Pseudomonas aeruginosa and non- tuberculous mycobacteria (NTM), can be extremely challenging, especially when these infectious agents co- infect the lungs of people with cystic fibrosis (CF) or other diseases. This application seeks to understand community interactions between P. aeruginosa and the antibiotic resistant NTM Mycobacterium abscessus. P. aeruginosa and M. abscessus are both found in the same environmental reservoirs and are co-isolated from individuals with CF and surgical site/soft tissue infections. Indeed, studies demonstrate that M. abscessus positive CF individuals are more likely to be infected with P. aeruginosa. Despite this, there is limited understanding of the interaction between these two important opportunistic pathogens. We recently investigated M. abscessus and P. aeruginosa interactions and reported several novel observations. P. aeruginosa potently antagonizes both rough and smooth M. abscessus variants, yet only when co-cultured in dual-species biofilms; antagonism was not observed in planktonic co-culture. Multiple P. aeruginosa strains exhibited antagonism, and P. aeruginosa-mediated antagonism was observed with another NTM strain, Mycobacterium smegmatis. Surprisingly, antagonism did not require known P. aeruginosa contact-dependent or -independent killing, motility, or oxygen/iron sequestration mechanisms. Thus, we hypothesize that inter- bacterial antagonism of M. abscessus by P. aeruginosa in dual-species biofilms is mediated by a novel antibacterial strategy. Aim 1 will utilize several complementary strategies to identify the mechanism of interbacterial antagonism. Aim 2 seeks to investigate M. abscessus and P. aeruginosa interactions under conditions that closely mimic those found in the host. The growing prevalence of M. abscessus infections and their recalcitrance to both host clearance and antibiotics highlight an urgent need to find new strategies to successfully treat these persistent infections. Successful completion of this proposal will advance our understanding of the novel P. aeruginosa antagonistic mechanism towards M. abscessus biofilms and provide novel biofilm targets that could be leveraged to augment antimicrobial efficacy in people with inflammatory and muco-obstructive lung disorders such as CF, chronic obstructive pulmonary disease (COPD), and non-CF bronchiectasis as well as with other NTM lung infections such as Mycobacterium avium.

NIH Research Projects · FY 2025 · 2024-05

Project Summary/Abstract: Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are problematic because of associated high treatment failures and elevated mortality rates. Staphylococci including MRSA are the major cause of medical device infections (MDIs) due to their strong capability to form biofilms. These bacterial biofilms resist removal by the host immune system and lead to antibiotic-treatment failures due to bacterial tolerance, development of antibiotic resistance, and limited antibiotic penetration. Vancomycin is the recommended therapy for MRSA MDIs and the antibiotic daptomycin is the primary antibiotic alternative to vancomycin for these infections; however, the development of daptomycin resistance especially post vancomycin therapy has been reported with increasing frequency. Dalbavancin is the so called “last line antibiotic”, a new lipoglycopeptide being used to treat MRSA infections. Our preliminary data furthermore have shown that neither daptomycin nor dalbavancin monotherapies are effective in eradicating high MRSA bacterial loads. In this context, the first aim of this project investigates the impact of a single dose of dalbavancin in combination with a single dose of daptomycin in biofilm state (in vitro pharmacokinetic/pharmacodynamic (PK/PD) model with components of prosthetic material for biofilm growth). This aim is particularly important for combating MRSA infections associated with MDIs in out-patient settings. Bacteriophages (phages) have been found to readily serve as anti-biofilm agents, and in some cases can also encode biofilm matrix hydrolyzing depolymerases. Therefore, the second aim proposes a single dose of dalbavancin adjunctive to a single dose of daptomycin (day 1) and daily administration of bacteriophage Sb-1 to eradicate MRSA infections in biofilm state (in vitro PK/PD models as stated in aim 1). We will use pharmacokinetic monitoring to achieve dose de-escalation of antibiotics in antibiotic-phage treatment. Collectively, this proposal will apply pharmacokinetic measurements, such as area under the concentration- time curve to achieve dose-de-escalation in phage-antibiotic treatment regimens. The proposed research is significant in the context of preserving current and future antibiotics and provides critical information regarding resistance prevention/re-sensitization using antibiotic-phage co-treatments. We will test our central hypotheses by evaluating the susceptibility of biofilm embedded MRSA to the various proposed antibacterial combinations and then perform in vitro two-compartment PK/PD biofilm models with humanized pharmacokinetics to optimize these novel therapies. We expect that through optimizing therapy of MRSA-biofilm infections, we will improve patient care and prolong the useful life of dalbavancin and daptomycin for the management of MRSA MDIs.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY/ABSTRACT Young adult alcohol misuse is an urgent, growing public health crisis, as young adults have the highest alcohol use disorder rates of any age group with rates increasing in young women. Interventions for this population are hampered by small effects, few options and lack of tailoring to salient risk factors. A recent review in Addiction argued that research informing interventions fails to account for the complexity of relationships among factors contributing to young adult alcohol misuse. We will address these needs by examining relationships between two cardinal etiologic risk factors: impulsivity and subjective response to alcohol (SR). Despite theoretical and biological links between them, little is known about relations between the two risk factors, or how associations between them may promote young adult alcohol misuse longitudinally and on a momentary or daily basis. Prior theory and evidence linking impulsivity and positive, rewarding alcohol effects have been primarily learning and expectancy based. Here, based on exciting preliminary results, we posit an inherent, biological link between impulsivity and high-risk SR (elevated stimulation and dampened sedation). This research is nascent with multiple gaps in knowledge. We will address these gaps by examining impulsivity pre-drinking, SR and alcohol use in a lab setting and via seven 10-day daily assessment periods over 2 years using ecological momentary assessment (EMA) (N=250, 50% female). Using self-report and both lab and mobile tasks, we will characterize 3 unique, established impulsivity domains: poor inhibitory control, delay discounting and negative urgency. SR will be assessed at successive breath alcohol levels using precision intravenous (IV) methods in the lab, followed by opportunity to self-administer more IV alcohol. We will also measure SR at roughly comparable, estimated blood alcohol levels via daily EMA methods. This design enables testing of SR early in a drinking event as a predictor of in-lab and daily alcohol use, along with alcohol use and consequences over time, plus SR’s potential role as a mediator of impulsivity/alcohol relations. Recent findings indicate daily changes in impulsivity predict subsequent drinking and consequences. These types of changes are challenging to capture with lab methods only. Daily measures also enable modeling of both person-level individual differences and daily, within-subject effects. However, there are no published studies relating daily impulsivity and SR measures. In this study, we will: 1) determine relations between lab-based impulsivity and SR; 2) determine relations between daily impulsivity and SR; and 3) relate impulsivity and SR to alcohol misuse longitudinally. We hypothesize impulsivity will relate to heightened stimulation and less sedation following alcohol and that SR will partially mediate relations between impulsivity and alcohol misuse. Evidence of links between specific impulsivity domains and SR longitudinally and on a momentary/daily basis will point to specific intervention targets to ameliorate two critical vulnerability factors for young adult alcohol misuse. Thus, we will: 1) identify mechanisms of alcohol action and 2) facilitate prevention and treatment research: two NIAAA priority areas.

NIH Research Projects · FY 2025 · 2024-05

Project Summary/Abstract Unilateral cochlear implantation is the standard treatment for adults with bilateral moderate to profound hearing loss, and it is estimated that up to 1.2 million Americans could benefit from cochlear implants (CIs).1 Bimodal hearing (CI in one ear and a hearing aid in the contralateral ear) is the most common CI listening configuration.2 Yet, an increasing number of patients are pursuing a second CI.3 Currently, there are no evidence-based criteria for when to recommend a second CI, resulting in a common clinical dilemma: should this patient continue using bimodal hearing, or should a second CI be considered (i.e., transition from bimodal hearing to bilateral CIs)? Previous attempts to provide evidence-based guidance on this clinical dilemma have focused on performance-based auditory tasks such as speech recognition;4-7 however, recent evidence suggests that patient report may have great utility when determining if bimodal hearing or bilateral CIs is right for a particular patient.6 Despite recent findings, the patient-reported advantages and limitations of each listening configuration have remained understudied, thus limiting the potential ability of patient-report in solving the abovementioned clinical dilemma. Our preliminary data demonstrates unexplained patient-reported differences between bimodal hearing and bilateral CIs, but that previously unknown information regarding the benefits and limitations of each listening configuration can be uncovered using qualitative patient interviews. Thus, the present study aims to determine the patient-reported advantages and limitations of bimodal hearing versus bilateral CIs using a mixed methods approach and two research designs (within-subject and between- subject). Aim 1 will compare bimodal hearing, bilateral CI, and bilateral candidates (bimodal patients scheduled to receive a second CI) using clinically relevant Patient-Report Outcome Measures (PROMs) and results of thematic coding obtained from qualitative patient interviews. Aim 2 will follow bilateral candidates through the preoperative (bimodal hearing) and postoperative (bilateral CI) phases using the same measures described in Aim 1 and will assess changes in patient-report across the two timepoints. Our preliminary data demonstrates the feasibility of this project and supports our central hypothesis, that patient reporting can offer valuable information about each listening configuration that can aid in differentiating the advantages and limitations of bimodal hearing and bilateral CIs. The long-term goal of this project is to equip clinicians with an evidence- based approach to counseling bimodal patients on the most beneficial listening configuration determined by individual needs and expectations. The proposed project will provide hands-on training in mixed methods research, advanced statistical analyses and interpretation, and help PI Lewis establish a unique line of clinically relevant research, thus preparing her for a post-doctoral position at an academic institution.

NIH Research Projects · FY 2026 · 2024-05

ABSTRACT This application, “Transfer RNAs as novel mediators in acute lung injury,” is submitted by Joseph S. Bednash, MD for a K08 Mentored Clinical Scientist Research Career Development Award. I am a physician-scientist in the Division of Pulmonary, Critical Care and Sleep Medicine at the Ohio State University (OSU). I am applying for this award for advanced training as a physician-scientist, studying mechanisms that contribute to dysregulated biology in ARDS and critical illness. The primary objective of my research plan is to determine how transfer RNA (tRNA)-derived fragments regulate host defense mechanisms in lung injury and to determine how TRMT1 (tRNA (guanine(26)-N(2))-dimethyltransferase), a tRNA methyltransferase and putative upstream regulator, modulates tRNA fragments to impact host defense. With cellular stress, tRNAs are fragmented into biologically active small non-coding RNAs, termed tiRNAs (tRNA halves) or tRFs (tRNA-derived fragments). TRMT1 methylates tRNAs, prevents tRNA fragmentation, and preserves protein translation efficiency. tRNAs, tRNA-derived fragments, and their control by TRMT1 has not been explored in ARDS and other critical illness syndromes. I present preliminary data demonstrating upregulation of tRNA fragments with bacterial endotoxin that promote cell death in macrophages. Further, I show that TRMT1 prevents tRNA fragments and preserves host defense mechanisms. The specific aims of this study are: 1) Determine how tRNA fragments contribute to immunopathology in ARDS and experimental lung injury, and 2) Define mechanisms whereby TRMT1 preserves inflammasome function in experimental lung injury. These studies will provide mechanistic insight into the role of tRNA fragments and their control by TRMT1 in lung injury and may identify novel strategies to augment host defense in ARDS. To accomplish these studies, I propose a training plan to acquire training in RNA molecular and cellular biology, training in microbial pathogenesis and immunology, and translational bioinformatics. To ensure success of my training and research plans, we have assembled a committed mentorship team, including i) Dr. Rama Mallampalli, Professor and Chair of Medicine at OSU, ii) Dr. Joshua Englert, an outstanding translational physician-scientist studying ARDS, iii) Dr. Federica Accornero, an expert in RNA modification in cardiac disease, and iv) Dr. Guy Brock, a biostatistician with expertise in translational bioinformatics. We have assembled a Scientific Advisory Committee to lend expertise, provide oversight, and evaluate progress, including i) Dr. Mallampalli, ii) Dr. Anuradha Ray, an expert in lung immunology, and iii) Dr. Ana Mora, an expert in mechanisms of lung injury and fibrosis. My work will be completed in the Division of Pulmonary, Critical Care, and Sleep Medicine at OSU, an excellent environment for my career development. With the guidance of mentors, the research environment at OSU, and support from the K08 Career Development Award, I am confident I can build my skillset and research program to support transition to independent R01 funding in the next five years.

NIH Research Projects · FY 2024 · 2024-05

PROJECT SUMMARY/ABSTRACT The sex gap in alcohol consumption is closing rapidly, due to alarming increases in alcohol consumption among young women. As such, there is an urgent need to determine the factors underlying sex differences in risk for AUD. Current addiction models propose three neurofunctional domains that drive problematic alcohol use and therefore serve as candidate sex-specific risk factors: executive function, negative emotionality, and incentive salience. Data from our lab and others suggest that poor inhibitory control, a key component of executive function, is a stronger risk factor for women than for men. Moreover, we have preliminary evidence that female drinkers show less engagement of neural circuitry underlying inhibitory control, and that this sex difference is influenced by circulating levels of estradiol. However, the degree to which hormonally-moderated sex differences in executive function extend to the negative emotionality and incentive salience domains, and how these sex differences influence current and future drinking is unknown. Here we will determine: 1) the neurobiological factors contributing to sex-specific risk for AUD in each of these three addiction domains and 2) the degree to which sex differences in each domain influence current and prospective drinking. Female drinkers will undergo fMRI to assess neural correlates of inhibitory control (i.e., executive function), negative emotionality, and alcohol cue reactivity (i.e., incentive salience) at three phases of their menstrual cycle: early follicular phase (low estradiol, low progesterone), late follicular phase (high estradiol, low progesterone), and mid-luteal phase (moderate estradiol, high progesterone). Male drinkers will undergo three fMRI scans at matched intervals. Immediately following each scan, participants will complete a session of free-access intravenous alcohol self-administration. We will then follow participants for 18 months to longitudinally assess changes in drinking patterns. We hypothesize that hormonally-moderated neural function underlying inhibitory control and negative emotionality will be stronger predictors of current and future alcohol consumption in women compared to men, whereas neural alcohol cue reactivity will be a stronger predictor for men. The project capitalizes on the unique skill sets of the PI (an Early Career Investigator) and a strong, collaborative investigative team. The innovative design will provide essential information regarding neural factors influencing development and maintenance of AUD, and, critically, how this risk is influenced by sex and fluctuations in sex hormones. Ultimately, this proposal is a crucial step in a line of research that will lead to the development of sex-specific prevention and treatment efforts for AUD.

NIH Research Projects · FY 2025 · 2024-05

Alzheimer’s disease (AD) is a major public health crisis. It is estimated that in 2022, 6.5 million Americans ages 65 or older lived with AD, and 5 million among the same population had mild cognitive impairment (MCI). By 2025, the number of AD cases will reach 7.2 million. The healthcare costs for individuals with Alzheimer’s or other dementias are substantial. The total payments in 2022 for all individuals with AD or other dementias were estimated to be $321 billion; by 2050, the annual payments for AD healthcare will total almost $1 trillion. However, resources are limited, particularly for underserved populations, and identification of cognitive impairment is often delayed so long that more effective treatments are underutilized. Therefore, expert panels have continued to stress the need for validated, brief, case-finding cognitive assessment tools, especially self-administered tests that allow for completely unsupervised administration and can accurately identify those with MCI. However, adding to the difficulties of AD diagnosis during the MCI stage are the significant disparities in the prevalence of AD and access to healthcare in racial/ethnic minority groups and other under-resourced and/or underserved populations. Black and Hispanic individuals are disproportionately more likely than White individuals to have AD, yet their socioeconomic disadvantages impede the early detection of MCI or AD. Thus, having easy access to low/no-cost validated and accurate self-administered cognitive assessments that can be taken in any clinic or non-clinic setting is highly consequential for the early detection of cognitive impairment. The goal of this project is to leverage the electronic Self-Administered Gerocognitive Examination (eSAGE), the metadata collected during eSAGE test-taking, the rich EHR data, and advanced ML techniques to develop such tools, which are particularly accessible to all individuals, including those socioeconomic disadvantaged, AD-vulnerable populations. To achieve the goal, we have three Aims. Aim 1 is to develop behavioral data and metadata, and ML methods to enhance the scoring and predictive ability of eSAGE. Aim 2 is to combine eSAGE test data and metadata/behavioral data with EHR data and develop new ML approaches to increase predictive capacity for cognitive status detection. Aim 3 is to test and validate the new ML- and EHR-enhanced eSAGE with the OSU Wexner Medical Center Memory Disorders Clinic, and with the underserved rural and minority populations in Ohio through outreach and tele-health (tele-cog) to evaluate their cognitive complaints. Successful completion of this project will produce a novel, translational, ML-enhanced eSAGE smart app and its integration within EHR systems, to improve our understanding of behavioral and clinical characteristics of cognition impairment, facilitate the identification of cognition impairment, and ultimately have a translational impact on AD identification and management. The project will enable a widely accessible, easy-to-use, and highly accurate cognitive assessment tool to be available for underserved, AD-vulnerable populations.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY Candidate and Environment: Dr. Rana’s career objective is to establish an independent research program aimed to develop therapeutic strategies to enhance respiratory function following spinal cord injuries (SCIs). This proposal has been carefully designed to supplement the candidate’s strong background in SCI neurobiology and respiratory neurophysiology with the acquisition of additional technical skills to study neuromodulatory therapies in rodents and will make her ideally suited to succeed on her career path. The University of Florida is an ideal place for Dr. Rana to achieve these goals since it is home to the Breathing Research and Therapeutics Center which brings together basic and clinician scientists devoted to understanding and addressing physiological challenges of respiratory motor control in disease and injury conditions. The core mentoring team consists of Dr. David Fuller (scientist) and Dr. Emily Fox (clinician-scientist), who are leaders in the field of respiratory motor control and SCI rehabilitation, and have a track record of successful mentees. Research: Respiratory complications are a leading cause of morbidity and mortality in the SCI population. Thus, strategies to target respiratory motor recovery are urgently needed. Transcutaneous spinal direct current stimulation (tsDCS) is a non-invasive neuromodulatory therapy that involves the delivery of a constant low-intensity current to target neural tissue, resulting in increased activation of spinal pathways and motor neuron excitability. However, the feasibility and efficacy of tsDCS to restore breathing following SCI has never been investigated. We have recently demonstrated that ampakines (allosteric modulators of α-amino-3-hydroxy-5- methyl-4-isoxazolepropionic acid (AMPA) receptors) enhance diaphragm muscle activation at acute and chronic stages of SCI. Thus, we hypothesize that 1) tsDCS can safely stimulate diaphragm motor output after incomplete cervical SCI, and 2) pairing tsDCS with low-dose ampakine promotes neuroplasticity, and therefore, is an effective respiratory neurorehabilitation approach. We will use a multidisciplinary approach, including controlled neurophysiological phrenic nerve preparations (Aim 1 & 2) and a comprehensive system to quantify diaphragm activity and overall ventilation in awake/unrestrained rats (Aim 3 & 4) to accomplish the following aims: 1.) Develop an effective tsDCS protocol to increase phrenic activation after cervical SCI; 2.) Test whether pairing ampakine therapy with tsDCS will promote sustained increases in phrenic output; 3.) Test whether tsDCS paired with ampakines can safely enhance diaphragm EMG output in awake rats with cervical SCI; 4.) Test whether a rehabilitation paradigm, consisting of daily tsDCS + low-dose ampakine therapy, can promote sustained recovery of diaphragm activation after cervical SCI. This application encompasses both mentored and independent phases. For the mentored phase of this application, a strong scientific community and support structure has been set in place. Together the co-mentors will guide the candidate in developing the necessary skills to complete this work and transitioning into a career aimed at developing effective strategies to mitigate SCI-inducted motor dysfunction.

NIH Research Projects · FY 2026 · 2024-04

OVERALL - ABSTRACT COVID-19 is a world-wide health problem caused by SARS-CoV-2 viral infection in the lung with long-term symptoms in at least one third of patients. Many COVID-19 patients suffer from silent or identified thrombi in major organs such as the lung and the brain and have increased occurrence of cardiac events. They also experience high levels of inflammatory cytokines collectively called cytokine storm. Combined, these reactions lead to organ damage and long- term sequelae of infection commonly termed Long-COVID. Our Program team will join forces to determine the host cell mechanisms underlying tissue damage in the lung and how SARS-CoV- 2 alters immune responses (Project 1), as well as in the brain and blood circulation (Project 2). Identification and targeting of host mechanisms that control the multi-organ inflammatory pathologies of COVID-19 will synergize with the targeting of cellular enzymes that control SARS- CoV-2 replication (Project 3). Together, our team will reveal and test novel therapeutic targets to collectively tame inflammation, neuroinflammation and thrombosis and to restrict viral replication. To achieve such a comprehensive overall goal, the three Projects by four Cores that will offer administration, biostatistical and bioinformatic support, animal models and purified viral strains, and relevant primary cell types with genetic manipulations to perform the planned experiments. Our Program will spearhead efforts to better understand the mechanisms of COVID-19 pathology in different organs and to identify novel drug targets to limit the severity of COVID-19 and the development of Long-COVID.

NIH Research Projects · FY 2025 · 2024-04

Abstract Nonsense Mediated mRNA Decay (NMD) degrades both aberrant transcripts containing Premature Termination Codons (PTCs) and “normal” transcripts with other specific features. Regulation by NMD is pervasive and estimated to impact around 10% of the transcriptome. UPF3 is a central NMD factor that bridges the mRNA bound Exon Junction Complex (EJC) with the rest of the NMD machinery and aids in PTC recognition. In mammals, there are two paralogs of UPF3, UPF3A and UPF3B, that have been documented to have both distinct biological roles and functions. We and others previously found that UPF3A can compensate for UPF3B in NMD, but is a weaker activator; moreover, this difference in activity is conferred by the “mid” domain. Moreover, overexpression of UPF3A, but not UPF3B stabilizes an NMD reporter mRNA. To understand the different propensities of UPF3 paralogs to stimulate NMD, we performed immunoprecipitation followed by mass spectrometry to identify their associated factors. In addition EJC and NMD components, we identified transcriptional regulators and nucleocytoplasmic shuttling factors to be among some of the most enriched factors in UPF3A and UPF3B immunoprecipitation. We also identified members of the nuclear transcription regulating Little/Super Elongation Complex (LEC/SEC); the LEC was previously identified as an NMD factor that promotes UPF3B association with the EJC. UPF3 is a nucleocytoplasmic shuttling protein, but its nuclear functions are unknown. For the K99 phase, I will investigate the impact of these nuclear functions on UPF3 deposition and NMD activity. I hypothesize that nuclear import is required for UPF3 function in NMD, and the LEC/SEC mediate nuclear UPF3 deposition onto nascent mRNPs. I will also investigate the functional differences between the UPF3 paralogs in human cells and in zebrafish development; I hypothesize that differential nuclear deposition underlies some of the functional differences between UPF3A and UPF3B, and their distinct roles in vivo arise from these functional, rather than expression differences. In zebrafish, Upf3a was implicated in the poorly characterized Genetic Compensation Response (GCR) via interaction with nuclear histone modifiers, and we found that this interaction is conserved in human cells. For the R00 phase, I will characterize the mechanism of GCR initiation from NMD-targeted mRNAs. I hypothesize UPF3A/Upf3a-stimulated NMD is required for GCR and UPF3A/Upf3a presence at these genomic loci determines GCR. Importantly, I have assembled a strong mentoring committee that will contribute their expertise to both my research training and professional development. This work will elucidate how the nuclear functions of the UPF3 paralogs impact NMD and how their functions contribute to developmental outcomes. Taken together, this work will deepen our understanding of how events in the nucleus and cytoplasm are coordinated to regulate cytoplasmic mRNA decay and its feedback to transcriptional regulation.

NIH Research Projects · FY 2025 · 2024-04

Project Summary: Immunotherapies have revolutionized the clinical treatment of melanoma; however, these treatment strategies have been ineffective in treating late stage and metastatic melanoma lesions with overall patient response rates below 50% illustrating an unmet medical need in melanoma therapy. We have previously demonstrated that mitochondrial metabolism played an important role in melanoma metastasis and hence, I sought to investigate the role of mitochondria in facilitating this aggressive form of the disease. Interestingly, our preliminary results indicate that specific deletion of mitochondrial complex I subunit Ndufs4 in tumor cells led to a dramatic anti-tumor immune response. Proteomic and metabolomic analyses of the tumor samples reveal that mitochondrial complex I inhibition induces an upregulation of proteins involved in antigen presentation, and a shift of choline metabolism from choline-sarcosine pathway to choline- phosphatidylcholine pathway. Tumor-Infiltrating Lymphocytes (TIL) analyses reveal a significant increase of NKT cells. However, the mechanisms by which mitochondrial complex I inhibition induces antigen presentation, metabolic flux shift and NKT cell activation are still waiting to be explored. Based on our encouraging preliminary results, I seek to further explore the mechanisms whereby mitochondrial complex I activity in tumor cells modulates immune response in tumor microenvironment by focusing on three Aims. In Aim-1, I will determine the mechanisms of how mitochondrial complex I inhibition enhances MHC-I dependent antigen presentation. In Aim-2, I plan to determine the mechanisms whereby mitochondrial complex I inhibition causes the metabolic shift, choline-betaine to choline-phosphatidylcholine, and its potential roles in NKT cell recruitment and activation. Finally, in Aim-3 I will evaluate the efficacy of combination treatment of immune checkpoint inhibitors with mitochondrial complex I inhibition in preclinical mouse melanoma models. While Aims 1 and part of 2 will be completed during the training stage, part of Aim 2 and the entire Aim 3 will be conducted during the independent phase of the award. The extensive training in different fields proposed in this application including proteomics, metabolomics and immunology will provide the tools to for me to become an independent researcher and study the mechanisms of which mitochondrial complex I regulates immune response in the tumor microenvironment. This training will be received in the vibrant scientific communities of Dana-Farber Cancer Institute and Harvard Medical School. This environment will expose me to the collaborations and discussions necessary for career development and future opportunities. Dr. Puigserver mentorship will be supportive to establish those connections and actively guide me in talk and manuscript preparation, student mentorship, experimental design, and career development. Together, the research and career development plans proposed in this application will strengthen my skills and competitiveness to become an independent researcher at a major institution.

NIH Research Projects · FY 2025 · 2024-04

Project Summary/ Abstract Traumatic brain injury (TBI) is a major cause of death and disability, with an estimated 1.5 million Americans receiving medical care each year for TBI. Patients with severe TBI require a decompressive craniectomy (DC), which is a lifesaving surgery to remove a large segment of the skull and relieve elevated pressure in the brain. The bone is then cryopreserved until brain swelling resolves (typically weeks to months), after which it is replaced in a cranioplasty surgery. Pathophysiological processes, including impaired revascularization of the cryopreserved bone, can lead to complete reconstruction failure, neurological deterioration, prolonged hospital stays, and increased economic burden. The foundation for this proposal is based on recent mouse model studies demonstrating that nonviral, nanotransfection based vasculogenic cell reprogramming can drive revascularization of the brain in ischemic stroke, as well as peripheral nerves in sciatic nerve injury, with improved functional outcomes in both instances. The current studies propose to utilize this technology for a new application: calvarial bone revascularization. Specifically, the aims of this proposal are to 1) Characterize and further develop a novel mouse model of autogenous cryopreserved cranioplasty after TBI and DC; and 2) Utilize fibroblasts nanotransfected with the transcription factors Etv2, Foxc2, and Fli1 (EFF-TNT) to induce vasculogenesis in autogenous cryopreserved calvarial bone in mice. Together, these studies will provide a mechanistic understanding of cryopreserved calvarial bone healing after decompressive craniectomy and explore an innovative approach to optimize healing. This proposal presents a five-year mentored career development program that will provide critical mentorship and position the candidate to transition from a primarily clinical role to an independent surgeon-scientist. The candidate is currently a tenure-track Assistant Professor at the Ohio State University. The outlined proposal builds on her previous clinical and research experience in calvarial reconstruction by integrating two new domains of expertise represented by her primary mentor Dr. Paco Herson and mentorship team: 1) Characterization of a novel animal model for studying pathophysiological processes in calvarial bone healing and testing therapeutic approaches (Dr. Ching-Chang Ko), and 2) utilizing an innovative, nanotransfection cell- based approach to revascularize bone and improve healing (Dr. Daniel Gallego-Perez). She is uniquely poised to carry out the proposed studies based on her training and background. She is firmly committed to a career as a translational surgeon-scientist and will have additional mentorship from Dr. Clara Lee to guide this aspect of her career trajectory. Completion of this comprehensive training plan will provide the candidate with the skills and experience necessary to become a leading independent investigator specializing in calvarial bone reconstruction after decompressive craniectomy.

NIH Research Projects · FY 2026 · 2024-04

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Adeno-associated viral (AAV) vectors are mainstream delivery platforms in gene therapy because of its safety profile and promising results in clinical trials. AAV has been successfully used to deliver transgenes to liver, heart, skeletal muscle, brain, and eye. Yet AAV-mediated gene transfer to adipose tissue is left far behind due to the low transduction efficiency and tropism of natural AAV serotypes. Our previous study demonstrates that an engineered hybrid capsid Rec2, transduces adipose tissue with superior efficiency than natural AAV serotypes. One caveat is that Rec2 capsid vector also transduces liver efficiently upon systemic administration. To mitigate off-target transgene expression in the liver, we have developed a novel dual-cassette vector design to severely restrict transgene expression in the liver. Although the dual-cassette design coupled with Rec2 capsid significantly improve the efficiency and selectivity of AAV-mediated gene delivery to adipose tissue, there are drawbacks such as the presence of Rec2 vectors in the liver and therefore bearing the risk of viral vector-associated toxicity; inability to suppress transgene expression in the liver if miRNA and shRNA are the transgene; the dual-cassette further reducing the cargo size. The weaknesses of existing technology highlight the need to generate new capsids with enhanced adipose tropism while eliminating liver tropism. Recently, we made several mutations in the Rec2 capsid to alter the tropism. Eight capsid variants (named LC.V1~8) have been generated, packaged, and tested in vivo. The capsid variant LC.V7, with three point-mutations, was found to possess exclusive adipo-tropism as it achieved substantial transduction in adipose tissue while completely eliminating liver transduction. Moreover, LC.V7 capsid vector did not transduce heart, spleen, muscle, kidney, or pancreas. Dosing experiment found that 4 x 1010 viral genome (vg) per mouse via intraperitoneal injection was sufficient for LC.V7 to transduce visceral fat and no transgene expression was detected in liver or any other peripheral tissues. Of note, 4 x 1010 vg/mouse is a low dose relative to systemic use of natural AAV serotypes often in the range of 1011 to 1012. In this project, we propose to develop the next generation of AAV vector platform for adipose gene delivery based on the novel LC.V7 capsid. In Aim 1, we will comprehensively characterize LC.V7 vector including biodistribution, administration routes, liver toxicity, and durability of transgene expression in lean and obese states. We will improve AAV genomic design to enhance transgene expression in adipose tissue. In Aim 2, we will develop LC.V7-based gene therapies for lipodystrophy. We will clone human leptin and human adiponectin cDNA sequence to the optimized AAV expression plasmid using a 2A sequence to express these two adipokines from one transcript and test the efficacy and safety of this gene therapy in two lipodystrophy models. This novel adipo-tropic AAV vector platform could provide a powerful tool for basic research and therapeutic purposes.

NIH Research Projects · FY 2026 · 2024-04

Project Summary/Abstract Ferroptosis is a unique type of programmed cell death that is induced by excessive lipid peroxidation. Emerging evidence suggests that ferroptosis represents a vulnerability in certain types of cancer that have acquired resistance to therapies. However, the signaling mechanisms that can be harnessed to promote cancer cell ferroptosis and its functional consequence on the tumor microenvironment (TME) reprogramming are poorly understood. In this Proposal, we aim to study a previously unrecognized anti-ferroptotic effect imparted by mitochondrial calcium signaling, which may also contribute to the establishment of immunosuppressive TME. Mitochondrial calcium uniporter (MCU) is a highly selective calcium channel that is localized to the inner mitochondrial membrane, which promotes the production of metabolite acetyl-coenzyme A (acetyl-CoA) by targeting the pyruvate dehydrogenase (PDH). In preliminary studies, we discovered that MCU-mediated acetyl-CoA generation blocks both cancer cell ferroptosis and antitumor immunity. We found that genetic ablation of Mcu (Mcu−/−) abolished acetylation of the glutathione peroxidase 4 (GPX4), a critical gatekeeper of ferroptosis, which correlated with impaired GPX4 activity and enhanced sensitivity to ferroptosis induction. Moreover, Mcu deficiency in cancer cells significantly blunted their growth and improved antitumor immune response. Therefore, blockade of MCU function may represent a promising therapeutic regimen for cancer. The goal of our proposed research proposal is to examine the function and mechanism of MCU- mediated acetyl-CoA metabolism on GPX4-controlled ferroptosis and its impact on antitumor immunity. We hypothesize that 1) decreased acetyl-CoA production in the absence of MCU sensitizes cancer cells to ferroptosis due to GPX4 hypoacetylation; 2) elevated ferroptotic cell death of Mcu−/− cancer cells promotes the production of type 1 interferon by intratumoral myeloid cells via the cyclic GMP-AMP synthase (cGAS)- dependent DNA-sensing pathway, leading to enhanced antitumor immunity; and 3) pharmacological inhibition of MCU by MCU-i11 synergizes with programmed death-1 (PD-1) blockade in limiting tumor growth. Representative syngeneic tumor models with high or low immunogenicity will be employed to examine the effect of MCU-mediated ferroptosis on antitumor immunity. Single-cell RNA sequencing analysis of tumor- infiltrating immune cells in response to MCU-i11 treatment will be performed to understand whether MCU inhibition promotes antitumor immunity. Results of these studies will provide novel insights into the induction and function of ferroptosis in rejuvenating antitumor immunity, which can potentially lead to the identification of new therapeutic targets in cancer treatment.

NIH Research Projects · FY 2025 · 2024-04

Project summary/Abstract Macrophages are considered a double-edged sword for tumor cells due to their plasticity in response to different environments. M1-like macrophages (M1) promote immune responses and inhibit tumor growth, while M2-like macrophages (M2) contribute to tumor progression by promoting angiogenesis and suppressing T cell responses. Tumor-associated macrophages (TAMs) predominantly consist of M2 macrophages, creating an immunosuppressive tumor microenvironment (TME). However, the mechanisms regulating macrophage polarization within tumors remain poorly understood. Tumor endothelial marker 8 (TEM8) has emerged as a key biomarker expressed on immunosuppressive cancer-associated fibroblasts (CAFs). Disruption of the Tem8 gene or blocking TEM8 protein has been shown to critically impede tumor growth in mouse models. My preliminary data demonstrate a striking increase in the M1 population within Tem8 knockout (KO) mice compared to wildtype (WT) mice. Notably, through an unbiased high-throughput CRISPR/Cas9 activation library screening, I recently identified TEM8 as a cell surface ligand for a macrophage receptor known as TEM8R, which plays a crucial role in regulating macrophage polarization. Moreover, TEM8 induces degradation of TEM8R through trans-endocytosis. Intriguingly, in vivo studies have demonstrated that monoclonal antibodies (mAbs) targeting TEM8, which effectively block the TEM8-TEM8R interaction, can inhibit tumor growth. Building upon these findings, my hypothesis is that TEM8 facilitates tumor growth by modulating macrophage differentiation in the TME through its interaction with TEM8R. To address this hypothesis, I will first elucidate the role of TEM8R in macrophage polarization within the TME by using the Tem8r KO mouse model I recently created. Subsequently, I will investigate the impact of the TEM8-TEM8R interaction on macrophage polarization and tumor growth using mAbs that specifically block this interaction, as well as a Tem8 mutant mouse model that is defective in this interaction. Furthermore, I will investigate the underlying mechanisms by which TEM8R regulates macrophage polarization and identify the factors governing TEM8R expression during macrophage differentiation. Additionally, I will explore the potential of anti-TEM8R mAbs and small molecule drugs that can disrupt the TEM8-TEM8R interaction as innovative approaches for cancer treatment and chemoprevention. Through unraveling the mechanisms by which TEM8, present on tumor-associated stromal cells, regulates TAM dynamics, these studies have the potential to advance our understanding of immunosuppression in solid tumors and facilitate the development of innovative and effective therapeutic strategies for cancer treatment and prevention.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY In the United States alone, in thirty years, prevalence rates of Alzheimer’s disease (AD) are projected to be at 13.8 million, with familial and societal costs estimated at $800 billion US dollars annually. With limited treatment options, there has been a renewed focus on targeting neurodegenerative, pathophysiological processes through behavioral and lifestyle-based interventions in the prodromal phase of AD. Presence of subjective cognitive decline (SCD)—referring to perceived persistent declines in cognitive functioning compared with previously normal cognitive status—has been identified as a potential preclinical stage of AD. Individuals with SCD show steeper declines in cognitive functioning later in life and have a higher rate of conversion to dementia. Moreover, SCD is also associated with the classical neural signatures of AD. Additionally, as individuals with SCD, by definition, show no objective indication of cognitive decline but are known to seek medical advice at memory disorders clinics, they make ideal candidates for prevention research. In the proposed application, our overall objective is to develop and test the feasibility of an entirely online, asynchronous mindfulness training program, and an active control group—internet-based Lifestyle Education (iLifeEd)—for targeting mind-wandering and plasma-based biomarkers of amyloid beta (Aβ) and tau pathology in adults with subjective cognitive decline. Mindfulness meditation involving the cultivation of purposeful and nonjudgmental attention to specific phenomena as they arise holds significant promise as an attention training platform. There is increasing support for engagement in mindfulness practices to reduce mind-wandering and enhance the executive control of attention, partially via mindfulness-induced alterations in connectivity of the default mode network with other large-scale networks. More recently, mindfulness has also been linked with higher volumes of the hippocampus, lower levels of amyloid burden and tauopathy in mid-life and older adults, suggesting a potential link between mindfulness training and AD biomarkers. However, clinical trials of mindfulness meditation are predominantly in-person, group-based MSBR programs that limit accessibility, and where online programs have been examined, studies show poor adherence and high attrition The proposed project is divided into two main objectives, wherein Aim 1 will iteratively develop and refine the iMBSR and iLifeEd protocols with a team of stakeholders, including psychologists, contemplative scholars, instructional design specialist, computer programmers, and focus group participants. Aim 2 is a Stage I pilot study, examining the feasibility of iMBSR and iLifeEd protocols, in an independent sample of 60 adults with subjective cognitive decline. Our main hypothesis is that iMBSR, carefully curated to include active components of the MBSR program, and designed to be entirely self-paced and online, will be feasible and acceptable to adults at higher risk for developing AD and other related dementias.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Every year, more than 15M babies are born too soon, resulting in more than 1M deaths. In fact, preterm birth (PTB) affects approximately 1 in 10 births across the globe, including 1 in 10 US births. Despite this, methods for the prediction and prevention of PTB are sorely lacking. Of note, more than 60% of PTBs are spontaneous (sPTBs), brought about by premature contractions or membrane rupture. The most common finding in sPTB is significant inflammation among the expectant mother. Yet, levels of circulating and even localized inflammatory markers during pregnancy fail to clinically predict who will versus will not progress to future sPTB. Similarly, antibiotics for asymptomatic infections and anti-inflammatories do not reduce risk for sPTB and are therefore not recommended for broad administration. To address such deficits, we developed a novel immunomonitoring method for use in prenatal care with the goal of predicting future sPTB and informing allocation of targeted, preventive interventions. In other words, we aimed to diagnose and address the immunopathologies driving sPTB risk. We have now shown that our novel prenatal immunomonitoring method can predict future birth timing in low-risk cohorts and future sPTB in moderate to high-risk cohorts, outperforming all available methods. In addition, our ex vivo experiments showed considerable inter-individual variation in the immunomodulatory effects of progesterone on immune function during pregnancy, which may explain why the drug is beneficial for some but not others. Thus, quantifying these immunomodulatory effects may provide a direct avenue toward targeted prevention of sPTB using a drug that is known to be safe during pregnancy. In the “Prenatal Immunomonitoring in Spontaneous Preterm Birth Prevention (PROMIS)” study, we propose to refine, expand, and clinically validate our prenatal immunomonitoring methods in a large, diverse clinical cohort. Our long-term goal is to save lives by predicting and preventing future sPTB. Our central hypothesis is that this can be accomplished using our novel prenatal immunomonitoring methods, which aim to diagnose and address the drivers of sPTB risk. To test this hypothesis, we’ll enroll a diverse cohort presenting for the prenatal care of singleton pregnancy, with early pregnancy patient-oriented data and biospecimen collection, mid-pregnancy cervical length measurement, and post-birth medical record review. We’ll test a prenatal immunomonitoring algorithm for the prediction of future sPTB. We’ll characterize the immunomodulatory effects of progesterone and examine the association between this profile and future sPTB. We expect the PROMIS study to produce novel insights into the pathogenesis, prediction, and prevention of sPTB. Importantly, risk prediction and targeted prevention go hand-in-hand, making advancements in one area dependent upon our capacity to advance the other. The potential impact of this study lies in its potential to identify sPTB risk using clinically feasible methods AND characterize risk phenotypes in a manner that allows us to address them. Thus, this project could provide unprecedented opportunity to predict AND prevent sPTB.

NSF Awards · FY 2024 · 2024-04

Water melting from glaciers flow on top of, through, and underneath glaciers, ultimately ending up in lakes or the ocean. Sometimes, this water does not take a direct path to the coast, but first enters near-shore lakes next to the glacier, called ice-marginal lakes, which can be dammed by ice. When the ice dams holding the water in these lakes fail, the water can rapidly drain to the coast over several days. The largest ice-marginal lake in Greenland is believed to be Lake Tininnilik, which recently drained into the ocean in 2021. The current low water levels mean that sediments in the lake are now exposed. This will be the first study to measure chemicals, nutrients, and microorganisms in the lake and sediments. It will also determine how lake storage changes the chemistry of water melting from the glacier. With continued climate warming, the amount of water stored in ice-marginal lakes is expected to increase, and determining the chemistry of Lake Tininnilik is important for understanding ecosystem change and carbon cycling in the coastal ocean following drainage. The project leadership includes three women, all of which are ethnic and racial minorities. As part of this project, a web-based virtual reality tour of Lake Tininnilik will be created for anyone to use. The tour will be important for other scientists trying to better understand the layout of the lake and will also be a teaching tool for the public. Because of erosion and weathering under ice sheets, subglacial waters are rich in macro- and micro-nutrients. These nutrient-rich waters can be directly discharged into the ocean or stored in pro-glacial lakes, including ice-marginal lakes. Lake Tininnilik is a large ice-marginal lake restrained by an ice dam along Sarqardliup Glacier in western Greenland. It drains approximately every 10 years into a local fjord, most recently in 2021, exposing previously inundated sediments. Preliminary work prior to the 2021 drainage shows that iron (an important minor nutrient for marine phytoplankton) is 10 to 100 times greater than glacial meltwater entering the ocean directly. The iron concentrations are also paradoxically high compared to other redox sensitive element concentrations. This project will collect and analyze water samples from different lobes of Lake Tininnilik and exposed sediments to address how ice-marginal lakes change the chemical and microbial composition and availability of nutrients for near-shore and open-ocean ecosystems. Sarqardleq Fjord, into which Lake Tininnilik drains, is an important source of fish for local indigenous populations, and this work will aid future studies seeking to understand how rapid drainage events may affect the marine food web. 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 2026 · 2024-04

PROJECT SUMMARY / ABSTRACT Choosing the more effective therapy is imperative for borderline-resectable pancreatic cancer (BRPC). BRPC is a subcategory of pancreatic adenocarcinoma (PDAC) (about 20% of the entire PDAC cases). BRPC contacts peripancreatic arteries and/or veins but has the potential to be successfully resected after downstaging with effective neoadjuvant therapy. Currently, there are two first-line therapeutic options for BRPC patients, nab- paclitaxel with gemcitabine and FOLFIRINOX. Overall response rates of these regimens are comparable (20- 30%), and there are no robust data favoring one over the other. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) has the potential as a non-invasive tool for early evaluation of PDAC response to chemotherapy. PDAC is typically hypo-perfused due to significant tumor sclerosis creating elevated interstitial pressure and consequently compressing tumor-feeding vessels. However, an effective therapy induces acute necrosis, reducing interstitial pressure and increasing perfusion. As DCE-MRI can measure tissue perfusion by monitoring the dynamic change of MRI contrast agents introduced intravenously, it can detect perfusion increase in responding PDAC before the morphological change of tumors. However, variability in quantitative DCE-MRI measurement remains a concern. We developed a perfusion phantom named P4 (Point-of-care Portable Perfusion Phantom) to reduce the variability in quantitative DCE-MRI measurement. The reproducibility of quantitative DCE-MRI measurement (e.g., volume transfer constant: Ktrans) of various abdominal tissues across three MRI scanners significantly increased after P4-based error correction (Intraclass correlation coefficient: 0.39 vs. 0.98). We demonstrated that quantitative DCE-MRI could be used to identify the early therapeutic response of PDAC after P4-based error correction. In our pilot study, DCE-MRI was applied for 20 PDAC patients before and 6-8 weeks after therapy initiation. The Ktrans of pancreatic tumors favorably responding to chemotherapy increased 84±26% (n=11) after P4-based error correction, while that of non-responding tumors did not (-7±11%) (n=9) (p<0.0001). We achieved 100% accuracy in differentiating between responding and non-responding tumors. We hypothesize that DCE-MRI-guided neoadjuvant chemotherapy will improve the negative margin (R0) resection rate for BRPC patients when the P4 is used for correcting errors in the imaging data. We propose to switch the first-line therapeutic regimen to the second one if the Ktrans in the tumor increases more than the threshold. We will compare the R0 resection rate of the group that received DCE-MRI-guided neoadjuvant chemotherapy with that of the historical control group that received standard-of-care treatment (primary endpoint). Also, we will determine whether quantitative DCE-MRI can be a reliable tool for assessing pancreatic tumor microenvironment after P4-based error correction using digital histopathology (secondary endpoint).

NIH Research Projects · FY 2026 · 2024-03

The lung is a major determinant of human health and respiratory diseases are a leading cause of morbidity and mortality among the elderly. The elderly population is one of the fastest-growing demographics in the US and worldwide. Our innovative T32 program in Biology of Aging and Pulmonary Diseases addresses a critical need to train biomedical researchers, physician scientists and future leaders in the links between aging and lung disease. We propose a multidisciplinary training program that will integrate basic mechanistic investigation of pulmonary disease and the biology of aging, with translational and clinical investigation of acute and chronic lung disease. The Division of Pulmonary, Critical Care and Sleep Medicine (PCCS) at The Ohio State University has been in a growth phase with the influx of investigators with well-recognized research programs and established track records of training. This new training program will be co-directed by Ana L Mora, MD, the Associate Director of research in the Davis Heart and Lung Research Institute and Rama Mallampalli, MD, the Chair of the Department of Internal Medicine (DOIM). An Executive Committee comprised of Jeffrey Horowitz, MD, Division Director of PCCS, and Mauricio Rojas, MD, Vice-Chair of Research in the DOIM will support Drs. Mora and Mallampalli in the administration of the training program. Five total positions will be available for MD and PhD scientists, with a 3-year structured, milestone-driven curriculum based primarily in laboratory research and complemented with research and career development retreats, translational core competencies, seminars, an academic survival skill conference series, workshops, and grant writing workshops. Our training plan is structured around individualized development plans that emphasize quantifiable outcomes including publications, career development awards, didactic courses and transition to research and academic careers. A dual mentorship training design will give trainees essential cross-disciplinary scientific and professional guidance in 6 areas: 1) Aging, Senescence and Metabolism, 2) Immunity and Host Defense, 3) Injury and Repair, 4) Environmental Exposures, 5) Therapeutics and Transplant, and 6) Biomedical Informatics. OSU’s environment for pulmonary training is unparalleled, with faculty in the Department of Internal Medicine-Division of Pulmonary Critical Care and Sleep Medicine, Department of Microbial Infection and Immunity, Department of Physiology and Cell Biology, Department of Surgery, Department of Biomedical Informatics, Department of Biological Chemistry and Pharmacology, Department of Pediatrics, Department of Molecular Genetics, Department of Chemistry and Biochemistry, and the Davis Heart and Lung Research Institute engaged in NHLBI-funded research across basic, translational, and clinical disciplines. This faculty, infrastructure, leadership, trainee pool, and unique scientific focus will address a critical pulmonary field and build a novel training program.

NIH Research Projects · FY 2025 · 2024-03

Significant vision problems (SVP), such as significant uncorrected refractive error, amblyopia, and strabismus are common in school-age children and may impede learning. Vision screening, a key public health tool to identify school-age children at risk for SVP, is mandated in 41 states, however it is not known which currently used screening test(s) effectively identify at-risk school-age children. Timely, appropriate eye care for children who are identified through screenings can improve quality of life, facilitate learning, and decrease the burden of lifelong vision problems, and its attendant economic costs. Currently there is great variation in state vision screening requirements and in approaches adopted among organizations involved in screenings. There is a need for evidence-based school-age vision screening guidelines and policies, however this will require much more robust data on the effectiveness of screening tests than are currently available. Effective screening tests must have high sensitivity and specificity to accurately identify those at risk for SVP while avoiding undue and costly over-referrals. Lay and nurse screeners also must be able to successfully administer the screening tests to children, interpret findings and make appropriate referrals. Limitations of prior screening studies in school-age children include: 1) examining only those referred, leaving the percentage with missed SVP unknown; 2) evaluating only 1-2 instruments without comparing available screening approaches concurrently; 3) including a wide age range, without subgroup analysis in the school-age population; 4) small sample size with limited range of SVP included; 4) not evaluating sensitivity to detect each type of SVP; 5) using definitions of targeted SVP that vary and may miss visually significant refractive error in school-age children; 6) not including newer technologies. Thus, valid full-scale, side-by-side comparisons between alternative screening tests are vital. There is also a paucity of information on how SVP affect quality of life in school-age children. Considering disparities in access to eye care after vision screening referral, greater understanding of the facilitators as well as barriers for obtaining needed care after screening referral is also urgently needed. This proposal aims to show the feasibility of a full-scale, multi-center, multidisciplinary study to compare the effectiveness of using traditional (visual acuity and stereoacuity) and technology-based vision screening tests for identification of school-age children at risk for SVP and in need of an eye exam. Before conducting the full- scale study, we must show the feasibility of enrolling children at each site, completing vision screening testing, and collecting eye exam results. We will also refine study procedures (e.g. training, certification, testing, data collection, data management, and analysis) and develop a Manual of Procedures and Protocol for the planned, full-scale UG1 study. We also will seek input from key organizations involved in conducting or setting standards for children’s vision screening to ensure that crucial questions, including equitable access to care, will be addressed in the full-scale study. The findings will help inform future screening guidelines and policies.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY Developmental and epileptic encephalopathies (DEEs) are severe early-onset seizure disorders with treatment-resistant seizures, developmental delay/regression, and long-term cognitive and motor impairment. SCN8A DEE, caused by gain-of-function, missense variants in the voltage-gated sodium channel gene SCN8A (Nav1.6), is characterized by infantile-onset seizures, mild to severe intellectual disability, significant developmental delay, and elevated risk of premature lethality. A serious comorbidity in SCN8A DEE is the prominent motor impairment, including severe hypotonia, movement disorders, and a large proportion of individuals that are non-ambulatory. Two mouse models carrying the patient mutations p.Asn1768Asp (N1768D) or p.Arg1872Trp (R1872W) recapitulated seizures and early death but did not exhibit motor impairment. Thus, insight into the mechanisms underlying motor impairment in SCN8A DEE has not previously been possible. We developed a novel conditional mouse model of SCN8A DEE with the patient mutation p.Thr767Ile (T767I). The T767I mouse is the first model of SCN8A DEE that exhibits significant muscle weakness and motor impairment. Our preliminary data suggests that functional motor units are reduced in Scn8a-T767I/+ mice. We hypothesize that the Scn8a-T767I mutation alters spinal motor neuron activity, leading to dysfunction of the motor unit and motor impairment in SCN8A DEE. Thus, we propose to use this unique mouse model to address the gap in our understanding of the pathophysiology underlying motor impairment in SCN8A DEE. We will use the T767I mouse to 1) determine how Scn8a-T767I affects motor unit structure, function and connectivity, 2) determine how expression of Scn8a-T767I in motor neurons affects neuromotor development, and 3) determine whether motor impairment can be prevented or reversed with administration of an anti-sense oligonucleotide that reduces expression of Scn8a in brain and spinal cord. Information gained from these studies will provide mechanistic insight into the pathophysiology of SCN8A DEE that will aid in the development of new therapeutic strategies for this severe disorder.