Ohio State University

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract Background: Substance use disorders (SUDs), which can involve illicit drugs, prescription drugs, or alcohol, are common and result in substantial morbidity and mortality. Unhealthy alcohol and substance use are among the leading causes of preventable deaths in the US, are increasing, and are among the top causes of the shortening life expectancy in the US. Screening for unhealthy substance use is recommended by the US Preventive Services Task Force and other national groups because it can lead to improved health outcomes, yet most primary care practices do not have a system to screen for and manage SUDs. Objectives: (1) to determine if practice facilitation (PF) can achieve rapid implementation of patient-centered outcomes research findings to improve screening for and management of SUDs in primary care and (2) to compare whether learning collaboratives (LC), performance incentives (PI), or their combination can significantly improve the implementation of screening for and management of SUDs in primary care compared with PF alone. Methods: The STop UNhealthy Substance Use Now trial (STUN II) is a multisite 2x2 randomized comparative effectiveness trial that will enroll 48 primary care practices. STUN II will evaluate the comparative effectiveness of the following strategies for the implementation of screening and evidence-based interventions for SUDs in primary care: PF, PF plus LC, PF plus PI, and PF+LC+PI. Assessments will be conducted throughout a 12- month intervention period and at a 12-month post-intervention follow up, to assess sustainment. In addition, a mixed methods evaluation will assess the association between implementation effectiveness and putative moderators (e.g., organizational readiness for change) and mediators (e.g., implementation climate). Potential Impact: The study will produce fundamentally important evidence about the comparative effectiveness of PF, PF plus LC, PF plus PI, and PF+LC+PI on uptake of evidence-based screening and interventions for SUD when delivered to primary care practices. It will also generate scientific knowledge about how contextual factors influence the effectiveness of the various approaches in promoting clinical practice and office systems changes in primary care settings. The results will inform health systems, policymakers, and primary care practices about optimal strategies for implementation of screening for and management of SUDs in primary care.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Although antiretroviral therapy (ART) is successful to block active replication of HIV, it does not completely eradicate the infection. HIV remains persistently infected and viral load can rebound after ART withdrawal, presenting a major obstacle for cure of HIV/AIDS. Investigation of host machineries that regulate HIV replication will help to improve the understanding of the mechanisms supporting HIV persistent infection. It will also provide new strategies to perturb host regulatory factors for eliminating residual HIV. This topic is especially relevant for the HIV-infected drug users, since drug abuse creates a profound impact on HIV infection, increasing the difficulty to manage HIV viral reservoirs. Our earlier effort includes the identification of novel host restriction factors specifically associated with a rare subset of HIV-infected individuals (<1%), termed elite controllers (ECs), who can maintain long-term control over HIV replication in the absence of ART. In a preliminary study, we performed RNA sequencing analysis and identified alternative splicing variants in cells from ECs, HIV-infected individuals undergoing suppressive ART, ART-naive HIV-infected individuals, and healthy controls. Differential gene expression patterns that are specific to ECs and may influence HIV resistance were identified, including alternative RNA splicing and exon usage variants of the CREM/ICER gene (cAMP-responsive element modulator/inducible cAMP early repressors). The knockout and knockdown of specific ICER exons resulted in significantly increased HIV infection. Overexpression of ICER isoforms decreased HIV infection. We also preliminarily showed that ICER isoforms are dysregulated in cocaine users. Together, these earlier studies confirm that CREM/ICER is a unique and novel host restriction factor suppressing HIV replication. We propose to comprehensively investigate their roles in regulating HIV infection, particularly for cocaine users. Furthermore, we also propose to identify other host gene isoforms that are dysregulated by cocaine use, which overall promote HIV persistent infection. Our central hypothesis is that certain host genes, including CREM/ICER, undergo profound RNA splicing to generate distinct isoforms in HIV-infected cocaine users and thus support HIV persistent infection in this population, which can be targeted to benefit HIV functional cure and mitigate HIV- induced inflammation. These studies include three related but independent aims. In Aim 1, we will determine correlations of CREM/ICER, HIV infection, inflammation, and cocaine use parameters. In Aim 2, we will investigate roles of CREM/ICER in mediating cocaine’s effect on HIV infection and inflammation. In Aim 3, we will identify novel gene isoforms in CD4+ T cells and monocytes dysregulated by cocaine use.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY / ABSTRACT If a facile method to site-selectively install prosthetic groups at internal sites in genetically-encoded RNA were available, then it would be possible to modify RNAs and ribonucleoprotein complexes (RNPs) in native structural and intracellular context, thus elevating studies on RNA folding, trafficking, lifetime, interactomes and regulatory pathways. The objective of this application is to test the extent to which compact U-rich internal loop (URIL) sites can be used as a general targeting motif in structured RNAs. If it were possible to selectively target the URIL motif with chemical probes, then juxtaposition of the URIL site with protein binding RNA motifs would enable tracking and chemical modification of ribonucleoprotein complexes (RNPs). This would enable elucidation of motif-specific RNA location and interactome by fluorogenic and proximity (biotin) labeling of URIL RNPs; such unbiased motif-centered interactome readout is not possible with existing methods. We hypothesize that appropriately modified, URIL-targeting bifacial peptide nucleic acids (bPNAs) could enable intracellular fluorogenic URIL (FLURIL) RNA tracking and proximity labeling of URIL (PLURIL) RNPs, respectively. Our proposed plan begins with the synthesis of bPNA probes, followed by rigorous in vitro and intracellular evaluation, optimization and validation with existing tools and known interactome partners. FLURIL RNP tagging will be benchmarked against MS2-labeling, the gold standard in RNA tracking. PLURIL tagging will be tested by its efficacy in identification of known RNPs. Further, we will test the extent to which URIL tags can be used to probe disease-relevant RNP biology in the intracellular context of amyotrophic lateral sclerosis (ALS), using patient-derived cells. Investigation of ALS pathology is a highly active area, with attention focused on two major forms: C9-ALS and Fus-linked ALS. While C9-ALS represents a majority of ALS cases, Fus-linked ALS is most commonly found in juvenile, aggressive early-onset cases; notably, the pathological mechanisms of these two forms appear to be distinct. Dysregulated RNP biology centered on C9orf72 RNA (C9-ALS) and U1snRNA (Fus-linked ALS) identifies these transcripts as prime substrates for URIL tag probes. The rigor in the prior research lies in the substantive preliminary and published data supporting intracellular fluorogenic and proximity labeling of URIL-RNPs. These data form a strong scientific premise for the impactful and unique application of motif-specific URIL-tagging as a broadly enabling discovery tool in ALS pathology and other RNP-centered diseases.

NIH Research Projects · FY 2025 · 2023-09

United States (US) pregnancy-related mortality (PRM) has more than doubled over the last two decades, with an additional 1.5-fold post-pandemic rise. It’s estimated that 80% of US PRM is preventable, yet rates remain high. Roughly half of US PRM occurs postpartum (PP) after hospital discharge. During this period, cardiometabolic and mental health conditions are precipitating factors in most deaths. Such data is particularly alarming considering that only 60% of patients receive healthcare during the PP year and few patients receive care that is adherent to evidence-based guidelines, particularly after pregnancy affected by a cardiometabolic or mental health condition. This is perhaps not surprising considering that patients consistently report that, “after you have [a] baby, it’s all about the baby and you don’t have time for yourself.” And providers consistently report that the obstetric to PP primary care hand-off is challenging. In collaboration with the Ohio Department of Medicaid, we designed a novel dyadic mother-infant PP primary care program targeted toward Medicaid-insured patients transitioning out of high-risk pregnancy – the Multi-modal Maternal Infant Perinatal Outpatient Delivery System (MOMI PODS). Dyadic care is a critical component of the model, with mothers and infants cared for in tandem throughout the PP year, and beyond. MOMI PODS was also strategically designed to 1) facilitate a coordinated obstetric to PP primary care transition, 2) promote tailored, evidence-based care informed by the obstetric history, and 3) integrate clinical and supportive care to concurrently address clinical and psychosocial needs. We have now established 8 MOMI PODS sites and delivered MOMI PODS to >300 dyads. Our compelling preliminary data shows that MOMI PODS is feasible and acceptable, with patients attending 95% of visits that systematically incorporate clinical and supportive care. In the MOMI PODS hybrid type 1 randomized controlled trial (RCT), we will evaluate the effectiveness of MOMI PODS in mitigating PP cardiometabolic and mental health risk, identify the biopsychosocial mechanisms linking MOMI PODS to PP health, determine if MOMI PODS promotes consistent PP care delivery, and identify strategies to improve implementation. Our central hypothesis is that MOMI PODS will mitigate PP risk for all by improving risk profiles and facilitating access to evidence-based clinical and supportive care. Thus, the MOMI PODS RCT represents a critical step toward establishing a consistent, scalable model of integrative PP clinical and supportive care that is capable of mitigating PRM risk for all. MOMI PODS has significant potential to serve as a foundational model of care appropriate for scale up and replication across a range of healthcare settings.

NIH Research Projects · FY 2025 · 2023-09

Acute myeloid leukemia (AML) is the most common adult acute leukemia. Molecular features such as cytogenetics and somatic mutations are essential components of risk stratification; used in daily clinical practice to determine treatment modality and intensity. Our knowledge of recurrent genetic AML-associated features, survival associations, subsequent genetic risk classification and clinical practice is informed by large-scale genomic studies performed over the past decade. Current clinical practice is predicated on the supposition that adequate testing has been performed and that genetic background should not interplay with the known AML-associated genetic and genomic landscape or with genes that associate with treatment response and/or that drive AML-genesis. However, our published and preliminary data reveal that constitutional genetic elements affect not only the frequencies and impact of known AML-associated gene mutations, but also multiple recurrent variants in genes thus-far not implicated in AML-genesis. We hypothesize that: a) known AML-associated drivers may carry different prognostic significance and might need to be adjusted in clinical risk classifications depending on genetic backgrounds, and b) some unrecognized molecular features are novel drivers of AML-genesis and/or therapy resistance. We propose to extend our initial studies of AML genomes and genomics to provide statistical confidence around frequently mutated genes, and a more precise, molecularly-adjusted risk and treatment stratification for AML patients. Next, we will focus on NPM1c, a genotype which currently is predicted to confer favorable outcome in AML patients. We will use cutting-edge single-cell multiomic assays to delineate bystander clonal mutations from potential driver mutations, then we will biologically test their roles in clonality/leukemia-stem-cell frequency and treatment response in genetically heterogeneous patients. We predict this approach to provide an improved characterization of the genetic and genomic landscape of AML, identify those variants with prognostic significance, and provide exemplars of heretofore unrecognized drivers of treatment response and survival. The overall goal is to enable precision oncology approaches which accommodate the effects of underlying human genetics in all patients.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY Marijuana smoking and vaping is on the rise in the United States with the increased legalization of recreational and medical marijuana. The main psychoactive component present in marijuana, and added to vaping preparations, is delta-9-tetrahydrocannabinol (THC). Even though the effect of marijuana smoking on obstructive lung disease can vary between studies, it is recognized that marijuana smoking is associated with increased cough, sputum production, and chronic bronchitis often associated with infections, but the mechanism remains to be determined. Our preliminary data show that chronic exposure of THC alone to primary human bronchial epithelial cells grown at air-liquid interface alters several components of the mucociliary function (i.e. ion channels, cilia beating), resulting in increased susceptibility to infections. Environmental pollutants, such as cigarette smoking, can alter gene expression. Unfortunately, some of the changes can persist despite cessation. The goal of this study is to determine the effects of THC-containing preparations (marijuana and vaping) on bronchial epithelium host defenses. This proposal is submitted in response to NOT-DA-20-031 “Transcriptomic, epigenomic, regulatory RNA or functional genomic research in substance use disorders”. Here, we propose to (1) determine whether chronic exposures to THC through marijuana smoke or vaping differentially alter the bronchial epithelium functions involved in host defense against microbial pathogens; and (2) identify the nature of transcriptomic changes which persist in bronchial epithelium after cessation from exposures to marijuana smoke or vaping. This comprehensive study will reveal the effects of THC in marijuana smoke and vaping preparations on airway homeostasis and identify new molecular targets to prevent infections and development of lung diseases.

NIH Research Projects · FY 2025 · 2023-09

Modified Project Summary / Abstract: Prior research has documented substantial differences in levels of physiological stress among adolescents, a period of particular importance from a developmental perspective with the potential to influence chronic disease risk in adulthood. The proposed project seeks to explore the role of exposure to multiple dimensions of everyday activity space locations – including area-level violence and perceptions of belonging – in contributing to everyday (real-time) perceptions of both physical and psychological unsafety and their contribution to explaining differences in physiological stress during adolescence (Aim 1). We will also consider the impact of activity space exposures, unsafety perceptions, and physiological stress as predictors of substance use and mental health outcomes (Aim 2). The project will collect aim-relevant data on a cohort of adolescents residing in the Columbus, OH metropolitan area employing an innovative two-stage design: The first phase will produce rich cross-sectional data (N=700) including standard survey items for youth and their caregivers; estimates of youth activity space exposures from self-reported routine activity locations combined with extensive contextual ratings and administrative data, physiological stress markers (cortisol and inflammatory), and health outcomes. The second phase will involve a supplemental intensive longitudinal data collection period taking place over a six-month period for a subsample of Phase 1 youth (N=300) selected using a novel exposure-based sampling strategy. Data collected during this supplemental phase will include an unprecedented combination of continuous high-resolution GPS tracking; ecological momentary assessments of perceived psychological and physical safety; continuously-sensed heart rate variability; and hair cortisol concentration and C-reactive protein measured at three additional time points. In addition to analyses of the novel data sources produced by the project, by drawing on data from the Adolescent Health and Development in Context Study (2014-2016) previously conducted by our study team, we will also examine recent changes in activity space exposures, safety and physiological stress, and their health consequences (Aim 3).

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract A growing literature suggests autistic adults feel pain in a different manner than non-autistic adults and may experience more persistent pain than the general population. As pain is often the first sign of injury or illness, individual differences in pain perception or communication can significantly impact diagnosis and treatment of many health conditions. Alternatively, when pain is not addressed, this can lead to a pro-nociceptive pain profile, where endogenous systems facilitate pain instead of inhibiting it, potentially contributing to persistent pain. Autistic adults often experience sensitivity in other sensory domains that could extend to pain. For example, hyper-responsiveness can include pain-like reactions to everyday tactile stimuli like tags on clothing. At this point, several studies have demonstrated increased pain sensitivity2,3, increased pain-related anxiety2, and altered neural responses to pain1 in autistic individuals. Yet, several important questions remain that present barriers to improving pain treatment in autistic individuals. In this project, we propose to address several gaps in our current understanding, building a framework to investigate pain sensitivity in autism. First, we will address pain assessment and expression of pain in autistic adults. Currently, there is no consensus recommendation on pain assessment in autism. It is not clear if autistic individuals express pain in specific ways that differ from non- autistic individuals and if better pain assessment tools would improve pain management. In Aim 1, we propose to test different pain scales in self-reporting autistic adults to determine reliability. Additionally, we will assess how pain ratings on these scales correspond to observable pain behaviors. Secondly, we will address potential factors that may predict persistent pain in autism. It is currently unknown if hypersensitivity to pain extends from a general sensory hypersensitivity for autistic adults. In Aim 2, we will explore individual profiles of sensory reactivity to identify important relationships between pain and sensitivity in other sensory domains. We will also explore relationships with social and emotional factors that may impact persistent pain. Lastly, in Aim 3, we will address potential brain mechanisms of pain sensitivity in autistic adults. Our previous work indicated that autistic adults have a different neural response to heat pain across somatosensory and affective brain regions, but the functional connectivity of these regions and the relationship between neural responses and pain reports is not clear. Overall, we hypothesize that autistic adults have difficulty communicating their pain, leaving them at a greater risk for mismanagement of pain. Additionally, we hypothesize that autistic adults likely have a pro- nociceptive profile increasing the magnitude of pain. This combination of difficult pain communication and a pro- nociceptive profile leaves autistic individuals at risk for persistent pain and is a detriment to their overall health. At the conclusion of this project, our work could inform how clinicians assess pain in autism (Aim 1), identify autistic individuals at most risk for persistent pain (Aim 2) and spark new mechanisms to improve pain management in autistic adults (Aim 3).

NIH Research Projects · FY 2025 · 2023-09

Project Summary A concomitant rise in adolescent social media use and decline in adolescent mental health over the last decade has raised the question as to the degree of relationship between these factors. To better understand this relationship, we examine how passive social media use (pSMU)—monitoring other people’s lives without engaging in direct exchanges with others—leads to poor mental health (Aim 1), when people engage in pSMU and how this coping strategy impacts mental health (Aim 2), and for whom pSMU is likely to undermine mental health (Aims 3) in a longitudinal study (n=400) in adolescents (ages 13-17; 50% Male/Female; 50% Black/White youth). There will be a 2-month focused study period. The initial and final 2 weeks of the 2-month study period will use ecological momentary assessment-based surveys (5 prompts a day) to measure pSMU, and to characterize affective responses to both pSMU and stress exposures the youth encounter in daily life. Objective pSMU will also be measured using an app installed on the youth’s phone in order to continuously measure pSMU over the entire 2-month intensive study period. We will measure stress exposures and responses in two novel ways. First, using GPS tracking we will determine the youth’s exposures to objectively stressful environments (i.e., high crime areas). Second, we will use GPS to trigger EMA prompts at locations youth reported as being stressful at baseline. The influences of these experiences on mental health trajectories (measured at weeks 0,2,4,6,8 and 20 will be assessed. Additionally, we focus on the relationship of pSMU to two physiological pathways that are responsive to social stress and influence risk for poor mental health: a) the parasympathetic nervous system, the function of which will be continuously measured using a self-charging wristband worn by the youth for the entire 2-month intensive study period to quantify shifts in heart-rate variability (HRV); b) and the expression of immune and other socially stress responsive gene pathways in blood cells sampled on week 0,2,6, and 8. This will provide the opportunity to determine the effects of pSMU on momentary affect, momentary HRV, immune related gene expression, a marker of general inflammation (CRP), and changes in depressive symptoms (Aim 1). We will also determine the role of stressor exposure (self-reported and GPS based) upon pSMU (Aim 2a) and the degree to which pSMU as a stress coping strategy moderates the effects of stressor exposure on affective response, HRV, stress related gene signaling pathways, inflammation, and mental health measures (Aim 2b). To examine individual differences in these effects, we will determine the degree to which gender is associated with increased pSMU (Aim 3a) and whether gender moderates responses to pSMU (Aim 3b). Finally, because our sample will be half Black youth, we will also determine if race moderates the relationship between GPS derived stress exposures (including GPS determined exposure to heavily policed areas or racially exclusive areas) and pSMU (Aim 3c).

NIH Research Projects · FY 2026 · 2023-09

Nicotine is the most important driver of tobacco use. The tobacco industry’s manipulation of nicotine in cigarettes and smokeless tobacco led to products that were more addictive, abused, and deadly. With “starter” products for young people and nicotine-naïve users and “mature” products for established tobacco users, the industry has historically manipulated the “levers” of nicotine concentration and form to increase tobacco use across all populations. E-cigarettes (ECs) and more recently oral nicotine pouches (ONPs) vary these nicotine dimensions signaling that the industry is again following this profit-driven strategy. Recent advancements in nicotine synthesis also makes nicotine isomer a new product lever that the industry can adjust to drive consumer uptake through changes in pharmacokinetics and misleading marketing claims. While the US Congress and FDA have imposed important regulations to dissuade tobacco use among young people, we argue that the regulation of nicotine itself may be the most effective strategy to achieve optimal population health. Like the tobacco industry, the FDA can pull these same levers via product standards and marketing regulations to adjust the appeal and addiction potential of ECs and ONPs. Through our theme, “Flipping the Script”: Using the Industry’s Nicotine Playbook to Maximize Public Health, the Ohio State University Tobacco Center of Regulatory Science (OSUTCORS) will be guided by the industry’s Consumer Response Model to understand how nicotine in ECs and ONPs are impacting product appeal, addictiveness, use patterns, and toxicity. We posit that regulation of nicotine’s three dimensions can dissuade young people, including non-users, from using ECs and ONPs but still provide a satisfying and less harmful alternative for adult tobacco users for complete switching. Project 1 (P1) and P2 will examine the influence of EC and ONP nicotine manipulations on product appeal, abuse liability, use, and toxicity; P3, using ONPs as an exemplar product, will examine the influence of nicotine marketing claims on appeal and product trial; and P4 will examine how young people respond to both nicotine product marketing and design characteristics in the natural environment. The projects are supported by the Market Monitoring Core that will provide critical real-time marketing and purchasing of products by venue and the Product Evaluation and Manipulation Core that studies products varied by nicotine dimensions with chemical and toxicological analysis. They also will be supported by the Administrative Core. The Career Enhancement Core will foster the training of students, postdoctoral fellows, and early career investigators in tobacco regulatory science. Together, the OSU-TCORS addresses the FDA scientific domains of Product Composition and Design, Toxicity, Addiction, Health Effects, Behavior, and Marketing Influences. The projects will provide the FDA with critical, first-of-its-kind data by conducting complementary interventional and observational studies to inform the Consumer Response Model as a tool to protect public health.

NIH Research Projects · FY 2025 · 2023-09

PROJECT ABSTRACT Diabetes is the most common cause of peripheral neuropathy, a condition that impacts over 30 million Americans. Often, this condition occurs as painful diabetic peripheral neuropathy (PDPN), which is particularly challenging to treat in the clinic, in part due to the lack of effective treatments and the risk of dependency for opioid analgesics. Spinal cord stimulation (SCS) is a treatment that stimulates nerve electrical activity in the spine, and is proven to be effective for pain conditions, including PDPN, for which SCS is now FDA approved through multiple device manufacturers. However, the mechanisms by which SCS improves pain are not yet known, and may involve promoting neural plasticity and small fiber nerve regeneration. We hypothesize that the electrical activity in stimulated spinal nerves with SCS treatment improves pain due to, or in parallel to, improvements in peripheral tissue innervation. This is based on decades of research evidence that electrical stimulation can promote nerve plasticity and regeneration. We will compare 3 PDPN treatment groups (N=15 each) across an 18mo longitudinal study: conventional medical management (CMM), SCS treatment, or SCS with a delayed activation of the device at 3mo post-surgery (to control for placebo effect of the surgery or device implantation). A team comprised of neuroscientists, neurologists, a neuromodulation surgeon, and computational/biostatistical experts will collect an 8-point dataset across this study timeline, in order to rigorously compare pain improvement scores with small and large fiber neuropathy assessments, quality of life scores, and metabolic parameters. The innovation of this study is further driven by the inclusion of a new functional measure of small fiber nerve electrical activity, using the Detecting Early Neuropathy (DEN) device, which employs a microneedle array to collect data on nerve activity up to 4mm deep from the skin surface. This is the first device capable of measuring small nerve fiber activity as a diagnostic test for small fiber peripheral neuropathy, such as diabetic neuropathy. Taken together, this study will provide important new data on PDPN improvements with SCS treatment and whether mechanisms include changes to small fiber tissue innervation or nerve activity.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY Hernia repairs are one of the most common surgeries performed in the United States, with over 350,000 ventral hernia repairs (VHR) performed yearly. Beyond the high health care system expenditure on VHR, patients report poor quality of life, post-operative pain, and time loss at work. Management traditionally includes surgical restoration of the abdominal wall, and short-term physician follow-up to ensure surgical site healing. Even once surgery has restored the abdominal wall, individuals post-operatively demonstrate major impairments of the musculoskeletal system including poor strength and function which contributes to reduced activities of daily living and a lower self-reported quality of life. Participation in physical therapy (PT) post- operatively may directly address and improve overall strength, function, and post-operative pain, in turn improving self-reported quality of life. There is a need to characterize recovery of function after VHR, specifically with the addition of objective measures of function to standard of care. Notably, we do not know the modifiable clinical characteristics of patients who will benefit the most from undergoing standardized post- operative PT. As a step towards this, we must understand the patient- and surgery-specific factors linked to a likelihood of a successful outcome following VHR. This mentored training plan will use data from a CMS Qualified Clinical Data Registry and an NIH/NIDDK funded registry-based clinical trial to understand the complex relationships between patient- and surgical-characteristics as they relate to post-operative function. Findings from this fellowship will provide (1) robust predictive models to understand characteristics of patients who have a good or poor self-reported quality of life 1 year after ventral hernia repair and (2) foundational evidence in understanding treatment response to physical therapy using objective measures of function. This fellowship proposes a paradigm shift of the current approach to hernia disease by considering objective and patient-reported measures of function of as a primary post-operative outcome. Meeting the objectives of this fellowship will afford the applicant the knowledge and skills required of an independent investigator and provide requisite preparation for a faculty position at a research-intensive university.

NIH Research Projects · FY 2025 · 2023-09

This proposal is for a five-year research program for Dr. Jon Wisler, an Assistant Professor in the Division of Trauma, Critical Care, and Burn Surgery. This proposal studies the mechanistic events and immunosuppressive clinical consequences of epigenetic methylation events that occur in survivors of sepsis. Dr. Wisler is a highly productive researcher in the fields of epigenetic regulation, sepsis, and clinical outcomes. This proposal couples the knowledge and skills gained during Dr. Wisler’s NIH K08 program relating to epigenetic regulation with direct application to clinical and psycho-social outcomes. Survivors of sepsis exhibit a profound degree of immunosuppression with higher levels of functional decline, depression, subsequent infections, and long-term mortality. To date, investigations related to his topic are fragmented and lack synergy. Jon’s research program seeks to unify multiple areas of investigation to improve the long-term outcomes of survivors of sepsis. His preliminary data identifies that patients with sepsis exhibit significant increases in DNA methyltransferase (DNMT) activity during sepsis. This results in profound gene silencing and immunosuppression. Additionally, we show that survivors of surgical sepsis exhibit numerous negative psycho-social effects that may represent the clinical effects of these epigenetically mediated immunosuppression events. Our intent for this application is to integrate the research efforts of Dr. Wisler and elucidate the deleterious biopsychosocial consequences of these epigenetic events coupled with in vivo assessments of longitudinal immune function and restoration. We hypothesize that molecular or pharmacological means to control DNMT function has potential benefits to patients with sepsis for boosting their innate immune function during the recovery phase of post-septic insult. Incorporating and coordinating these areas of research will greatly improve our understanding of these epigenetic events and provide a unified analysis of mechanistic, translational, and clinical outcomes. Under the R35 program, Jon seeks to integrate cutting-edge laboratory-based investigations and therapeutic testing with patient-based assessments including time-course based immunologic dysfunction and altered clinical outcomes. Post-sepsis immunosuppression is an often diagnosed but untreated consequence of sepsis survivorship. This program will establish the time course, functional effects, and avenues of interventions to treat the underlying epigenetic events involved in this immunosuppression. This will generate paradigm shifting treatments for a disease process with significant clinical impact.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY / ABSTRACT Bladder cancer (BC) is the tenth most diagnosed cancer in the world, and is strongly linked to environmental chemical exposures including benzo[a]pyrene (BaP). BaP is a polycyclic aromatic hydrocarbon (PAH) and widespread pollutant found in automobile exhaust, tobacco smoke, and charred food, but the mechanisms underlying the links between BaP and BC remain unclear. Additionally, not all individuals exposed to BaP develop BC, indicating that there are additional undefined or individualized risk factors associated with BC. While human cells can metabolize BaP, host-associated microbes can too. Moreover, host-associated microbiomes, like host genomes, are individualized, which could drive differences in host responses to BaP exposure. However, the role of microbes in BC, and specifically, how the gut and urine microbiome modulate exposure to BaP, is largely unexplored. There is a critical need to evaluate chemical-microbe-host interactions in relation to high-risk chemicals – like BaP – on bladder health. The objective of this application is to define microbial metabolism of BaP in vitro and in vivo and asses the effects of this metabolism on the urothelium. BaP along with its metabolites are excreted in urine, have been linked to BC, and can be metabolized by microbes. However, it is unknown if microbial metabolism of BaP is occurring in vivo and how the microbiome may influence this metabolism. Our overarching hypothesis is that microbial metabolism of BaP is playing a role in bladder carcinogenesis. Specifically, we predict that urine and stool microbes can either “toxify” or “detoxify” BaP, with “toxifying” metabolism defined here as that which yields secondary metabolites more damaging or carcinogenic to host cells. In our first aim, we will characterize metabolism of BaP in vitro by urothelial cells and by gut or urinary tract-associated microbes. Host cells and bacteria will be grown in the presence or absence of BaP, separately and together, and we will apply targeted metabolomics (GC-MS) to quantify BaP metabolism. In the second aim, we will characterize metabolism of BaP in vivo through the gut and urine microbiome and metabolome of mouse models with differing microbial community profiles. In aim three, we will evaluate urothelial response (toxicity and inflammation) to BaP exposure in vitro and in vivo using samples from the first two aims. The proposed research is innovative because it represents a substantive departure from the status quo by defining chemical-microbe interactions and their direct impact on the bladder epithelium. This approach will create novel opportunities for bladder health and BC management through microbial community manipulation and engineering. Upon completion of the proposed aims, we will have identified which microbes can metabolize BaP, how they do so, if this metabolism is occurring in vivo, how differing microbial communities influence this metabolism, and how this affects urothelial response. This research is significant because it will establish a novel chemical-microbe-host framework for evaluating chemical safety, bladder health, and BC development.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT Pregnant and lactating people remain therapeutic orphans as they are excluded from the vast majority of clinical drug development and therapeutic trials. Moreover, current practices in drug evaluation in pregnancy have been hindered by the lack of effective biomarkers and innovative study designs. There is a need to develop novel placental-specific biomarkers in order to assess placental function and response to therapeutics, as a way to inform on their safety and efficacy. An example of these novel biomarkers is placental (fetal) specific extracellular vesicles (EVs). Recent advances in characterizing the cargo content of these EVs demonstrated their potential to be used as placental biomarkers. We have shown using funding support from the MPRINT that fourteen proteins found in EVs significantly correlated with aspirin use (FDR<0.1) in at-risk people, but that more power is needed to confidently assess the relationship with aspirin dosage and pregnancy outcomes. In addition, prior studies showed that aspirin affects endothelial and trophoblast cells, thus potentially modulating exosome derived from these cells and their cargo contents. Leveraging our prior proof of principle success, previously collected, and ongoing collection of maternal plasma from people at-risk of preeclampsia (PE) receiving 81mg or 162mg of aspirin daily and low-risk people at the Ohio State University, we are submitting this application with the overarching goal to develop a novel platform to augment the MPRINT resources using exosome profiling, as novel biomarkers, to monitor placental mediated adverse pregnancy outcomes and response to therapeutics. For this, we will use PE as a hallmark of these outcomes and aspirin as the therapeutic agent to show case the utility of the platform. In this study, we will test the hypothesis that differential expression of EVs proteome cargo is associated with placental and pregnancy health in response to aspirin treatment at different gestational periods. We will test the following specific aims: 1) Validate our preliminary proteome data and model using larger cohort from additional biobanked samples of at-risk people receiving 81mg or 162 mg aspirin, characterize fetal specific EVs (placental alkaline phosphatase [PLAP] +ve) isolated from maternal plasma, and determine the differences in between the maternal vs. fetal EV cargo difference with aspirin treatment; and 2) Determine the changes associated with EV proteome profile (maternal and fetal) in at-risk people receiving aspirin prospectively and correlate with clinical outcomes (development of PE) and angiogenic and inflammatory biomarkers associated with PE. This project has the potential to play a seminal role in the development of a novel platform for therapeutics research in pregnancy. This translational research platform will substantially add to the MPRINT Network repertoire and be available for scientific community through the already established relationship of the team with other networks such as the Maternal Fetal Medicine Units Network, MPRINT, and the Foundation of the NIH.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Bladder cancer is one of the top 10 most frequent cancers, yet the most underfunded cancer by NCI based on funding dollars and ratio of funding/mortality. There has been no significant improvement in overall survival and prognosis over the last thirty years except for the recent development of immunotherapy. After initial diagnosis and treatment, over 60% of the BC patients will relapse within two years and progression into advanced stages in up to 25% of patients. Therefore, almost all patients will need long-term expensive cystoscopy which makes bladder cancer the costliest cancer (per case) of all cancer types. If a sensitive but less expensive method to detect cancer were available, it would improve the treatment outcomes and decrease the cost. Dr. Xuefeng Liu (PI) and his team recently invented a robust method of conditional reprogramming (CR) technology to establish patient-derived cell models from cystoscopy or needle biopsies and urine samples of BC patients. The overall success rate was near 100%. These urine cancer cell cultures (UCCC) provide a simple, non-invasive, comfortable, reliable method to detect BC recurrence, and represent a living biomarker to predict patients’ response to chemotherapy and targeted therapy. Several cancer centers and laboratories including PI’s lab are actively involved in clinical trials to validate clinical utility of the UCCC in BC clinics. Since a variety of patient- context and collection, storage and transportation of urine samples to laboratories for cell cultures affect enrichment and viability of cancer cells in urine samples, it is urgent needed to evaluate pre-analytical factors of urine samples for UCCC and optimize these non-Invasive and living liquid biopsies as a routine approach in monitoring recurrence and prediction of response of BC patients. In this application, we propose to address the gaps by extending the work done by our team members on evaluating effects of sample processing protocols on UCCC analysis, to further investigate effect of patient-specific context. In this application, we will first further optimization of UCCC system to establish a culture kit for clinical applications, specifically we will optimize and simplify conditions with co-culture kits, conditioned medium (CM), hypoxic condition for maximal efficiency and robustness. Second, we will establish a Standard of Procedure - SOP- for urine collection and transportation in clinics: we will test pH, temperatures, minimal cell number, a cocktail with anti-biotics and Y-27632 affect UCCC cultures and success rate. Last, we will determine patient-context factors that affect UCCC cultures: we will evaluate how urine timing (days before surgery, and after surgery, early morning, noon, night, drinking water before urine collection, etc), urine volume affect cell viability and UCCC. Last, we will optimize a self- sampling procedures for BC patients. The overall goal is to expedite UCCC clinical assay development through evidence-based standardization of urine collection and handling practices, potentially approved by FDA as a standard method for bladder clinics.

NIH Research Projects · FY 2025 · 2023-09

Sarcopenia, the age-related loss of skeletal muscle mass and function, predicts for morbidity, mortality and excessive healthcare costs. Other than exercise, lifestyle therapies for slowing the progression of sarcopenia in older men and women remain elusive. Mitochondrial dysfunction is a common mechanistic link between aging and many pathological factors contributing to sarcopenia. We propose to test whether supplementing the diet with an oil rich linoleic acid (LA; 18:2n6), the preferred fatty acyl constituent of cardiolipin, improves muscle strength and physical mobility. In mice, dietary LA-rich oil increases LA-rich cardiolipin (e.g., 4-LA- cardiolipin) and mitochondrial respiration. Therefore, the effect of LA-rich oil to increase 4-LA-cardiolipin and mitochondrial respiration in older people with sarcopenia will be measured evaluated. Because LA is a required nutrient for humans, e.g., an essential fatty acid that must be attained in the diet, dietary sources are the sole method to replenish LA for cardiolipin, the signature phospholipid of inner mitochondrial membranes. In a phase II double-blinded, randomized controlled trial, this study will test the effect of LA-oil supplementation to change muscle strength, physical mobility, 4-LA-cardiolipin and mitochondrial function. Identifying a mechanistic link between LA and mitochondrial capacity could aid in the development of mechanistically targeted therapies to slow or stop the progression of sarcopenia as well as other mitochondrial-related conditions that develop in older adults.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY We aim to answer the signature question of paramedic care; “What is the best way to manage the prehospital airway in critically ill children?” Cardiac arrest, respiratory failure, and major trauma are devastating critical conditions in children. Resuscitation from critical illness requires skillful airway management to optimize delivery of oxygen to the lungs, preventing irreparable damage to the brain and heart. As the first to provide critical care, prehospital paramedics often perform life-saving airway management on critically ill children. The most common prehospital airway management techniques (bag-valve-mask ventilation (BVM), endotracheal intubation (ETI), and supraglottic airway insertion (SGA) have important trade- offs between risks and benefits; the best approach is unknown. We propose the novel Pediatric Prehospital Airway Resuscitation Trial (Pedi-PART) to determine the best strategies for prehospital airway management in critically ill children. Pedi-PART will immediately influence paramedic practices and have a lasting impact upon the health of children internationally. We will determine the relative effectiveness of three airway strategies ([BVM-only], [BVM+ETI], [BVM+SGA]), accounting for variations across disease (cardiac arrest, trauma, respiratory failure) and age categories, and we will apply adaptive techniques to minimize the required number of patients. Our aims are: AIM I: Prepare the Pediatric Emergency Care Applied Research Network (PECARN) to execute the innovative Pedi-PART trial. PECARN is the nation’s premier pediatric emergencies research network. We have assembled an alliance of over 60 EMS agencies from 10 communities associated with PECARN and their ongoing Pediatric Dose Optimization for Seizures in EMS trial (PediDOSE - U01-NS114042). AIM II: Compare the effectiveness of prehospital BVM, SGA, and ETI airway management strategies upon pediatric cardiac arrest, major trauma, and respiratory failure outcomes. We will execute the trial in two sequential stages. Stage I: Determine if [BVM-only] or [BVM followed by SGA] results in better ICU-free survival in critically ill children with cardiac arrest, major trauma, or respiratory failure. Stage II: Determine if [winner of Stage I] or [BVM followed by ETI] results in better ICU-free survival. AIM III: Compare the safety of prehospital BVM, SGA, and ETI airway management strategies in pediatric cardiac arrest, major trauma, and respiratory failure. In Stage I, we will determine if [BVM+SGA] results in fewer prehospital and hospital adverse events compared with [BVM-only] in critically ill children with cardiac arrest, major trauma, or respiratory failure. In Stage II, we will determine if the winner of Stage I results in fewer adverse events compared with [BVM+ETI].

NIH Research Projects · FY 2025 · 2023-09

Out-of-hospital cardiac arrest (OHCA) is a dynamic process that requires new interventions to improve outcomes. End-tidal carbon dioxide (EtCO2) measurement is a tool that is widely recognized, easy to use, and can potentially provide real-time insights into ongoing resuscitation efforts; however, it has yet to be applied to individualized medicine. Our overall hypothesis is that integrating EtCO2 capnography into OHCA resuscitation will improve outcomes. Using innovative signal processing and machine learning methods, we will identify a wide range of resuscitation quality characteristics over resuscitation, their relation to individual patient characteristics and predictability of OHCA outcomes. These goals will be accomplished via the following aims: Aim 1. Determine the influence of resuscitation interventions on real-time physiologic dynamics and outcomes in OHCA. Aim 2. Establish the influence of individual patient characteristics on the real-time physiologic dynamics and OHCA outcomes. Aim 3. Develop a novel cardiac arrest resuscitation strategy based upon real-time individualized physiologic dynamics. We will create a large repository of cardiopulmonary resuscitation process data encompassing data from 6841 adult OHCA. This work will define intra-arrest EtCO2 dynamics over resuscitation to allow for the development of guided resuscitation efforts, and the resultant data will provide a solid foundation for future hypothesis-driven research. Dr. Nassal’s training plan encompasses both formal didactics and experiential training with experienced mentors and collaborators that will develop a skillset in both signal processing and artificially intelligent driven algorithms. The team has extensive experience in using machine learning and multimodal signal processing for classification and predictions in resuscitation. This training program will develop a unique skillset in advanced cardiac signal processing; artificial intelligence, machine learning processing; and expertise in the application of these skills to develop dynamically guided resuscitation strategies that few other physician-scientist possess.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ ABSTRACT The central premise of precision medicine is that an individual’s unique physiologic characteristics play a significant role in disease vulnerability and response to specific therapies. To make progress towards precision medicine in psychiatry it is imperative to move beyond group-based studies that disregard individual variations as noise, and instead interpret individual variations in the context of the normal-range of biological systems, which is consistent with the Research Domain Criteria [RDoC] paradigm. In response to the Notice of Special Interest (NOSI) regarding the use of Human Connectome Data for Secondary Analysis we propose to use normative modeling (‘brain growth charting’) techniques to generate a comprehensive reference atlas for brain microstructure across the lifespan that contains age and sex based normative information for individual brain regions using publicly available connectome data (>50,000 individual datasets, age 10-100 years). We chose to target brain microstructure because postmortem and neuroimaging studies have implicated microstructural involvement in virtually all major psychiatric disorders. We will also create a web-based ecosystem that contains the relevant workflow components, including an option to calibrate models to new datasets. In addition to publishing final models and code as Jupyter notebooks, we will design a graphical user interface to reduce barriers for neuroscientists with limited experience in command line coding and to maximize the impact of our work. Taken together, this approach will provide a problem-agnostic, quantitative framework for characterizing variations at the individual level and give us a paradigm-shifting opportunity to advance clinical translation, where information of an individual patient’s brain microstructure can be leveraged to forecast the probability of a psychiatric condition, clinical outcomes, or response to treatment. We will showcase the value of this approach for psychiatric applications by determining the extent and clinical relevance of individual microstructure deviations in early psychosis patients. We chose this patient population because it is genetically and phenotypically heterogeneous, confounds of disease chronicity and antipsychotic medication exposure are limited, and because we have found that subtle microstructural abnormalities are already present at illness onset and associated with clinical and outcome variables. Using publicly available connectome data, we first will assess the extent of individual microstructure variations and assess heterogeneity in microstructural deviations from the norm in this patient population. We will then test the relationships between individual microstructure variations and key clinical variables, and determine if individual microstructural variations can be used to identify clinical subtypes. This constitutes a pivotal step towards precision psychiatry, where patients are conceptualized based on their unique physiologic characteristics, and insights from individual imaging signatures guide medical decision making.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Stringent regulation of inflammation during infectious and non-infectious diseases is critical for limiting tissue pathology while promoting disease resolution. The dead-box helicase family members known as retinoic acid- inducible gene I-like receptors (RIG-I-like receptors, RLRs) play a critical role in recognizing self and non-self nucleic acids in the cytosol of host cells. Dysregulation of RLRs and their downstream interferon (IFN) and inflammatory signaling cascade can manifest as autoimmunity or as defects in antimicrobial responses. Despite the critical importance of RLRs, the function of these receptors and their associated molecular pathways in different cell types, remains an important gap in our understanding of tissue homeostasis versus diseased states. As an independent investigator, my studies now focus on how emerging non-canonical functions of RLRs and IFN regulate inflammation in the context of pulmonary pathogenesis and immune cell programming, two critical areas in which better understanding of RLR-associated signaling could lead to new strategies for treating infectious and non-infectious inflammatory diseases. Our preliminary studies have found that atypical induction of the RLR RIG-I using a synthetic agonist leads to activation of immune cell programming genes rather than IFN induction. Moreover, we have identified new roles for type III IFN (IFN) in pulmonary tissue repair following pathogen-induced damage. My research program can be defined with three thematic goals: 1) elucidate how RLRs and IFN pathways contribute to tissue homeostasis, 2) determine whether RLR and IFN pathways are differentially activated in a strategically-selected set of cell types in damaged vs adjacent tissues, and 3) determine the distinct contributions of RLR and IFN in controlling non-infectious or infection-mediated inflammation and resolution. We have developed first-of-their-kind mouse models to eliminate expression of the RLRs RIG-I, MDA5, and their downstream signaling adapter MAVS, as well as type I and type III IFN signaling pathways in temporal and cell-specific fashions. These tools will allow us to use cutting edge high-parameter spectral cytometry, along with spatial transcriptomics, to define the most relevant inflammatory pathways deployed by individual cell types in the context of their natural tissue microenvironment. Ultimately, our studies will improve understanding of cell-intrinsic regulation of nucleic acid sensing pathways, and will provide strategies for differentially targeting each pathway to maximize therapeutic benefit while minimizing adverse side effects.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY As the SARS-CoV-2 pandemic continues, a better understanding of the factors that regulate broadly protective immunity is needed. Interferon-lambda (IFN-λ) mediates antiviral protection against SARS-CoV-2 without causing overt pathology and exacerbated disease. As such, recombinant IFN-λ is in clinical trials as a therapeutic for COVID-19. However, the endogenous role for IFN-λ in regulating infection and functions in preventing disease beyond antiviral programming during SARS-CoV-2 remain unknown. Our preliminary data show that mice lacking the IFN-λ receptor (Ifnlr1-/-) have increased illness and viral burden during SARS-CoV-2 infection without alteration of canonical antiviral gene induction associated with IFN. Instead, IFN-λ positively regulates the induction of CD8 T cell immunity, a cellular immune response critical to mediating protection against virus infection when neutralizing antibody responses are avoided. We further identified upregulation of cell cycle repair and proliferation genes associated with fibrosis in Ifnlr1-/- mice. This proposal will investigate these newly identified non-canonical functions of IFN-λ in regulating immunity against SARS-CoV-2 with the goal of informing therapeutic use of this cytokine.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT DNA cytosine methylation is a central epigenetic modification in mammalian cells. Aberrant DNA methylation has been linked to gene dysregulation and genomic instability, resulting in cancer and other diseases. Ten- Eleven Translocation (TET) is the essential enzyme family responsible for DNA demethylation to ensure the proper maintenance of the methylome. TET enzymes oxidize 5-methylcytosines (5mC) into mainly 5- hydroxymethylcytosine (5hmC), an intermediate for DNA demethylation and a potential epigenetic mark. Germline deficiency of TET2 is associated with immunodeficiency and childhood cancers. Notably, TET2 is one of the most frequently mutated genes in hematological cancers, demonstrating the importance of TET in multiple biological processes. We and others have shown that TET enzymes are essential for cell differentiation by regulating super-enhancers and inducing the expression of lineage-specific genes. However, the mechanism by which TET enzymes facilitate the permissive chromatin environment for gene activation remains unclear. The long-term goals of the research program are: (1) to comprehensively understand how TET enzymes and 5hmC promote chromatin accessibility; (2) to investigate the crosstalk between TET and histones; (3) to understand the consequences of temporary perturbation of epigenome. To accomplish these goals, we will use precision epigenome editing to manipulate and monitor epigenetic modifications of cis- elements regulated by TET and DNA methylation. We will develop novel animal models to study the impact of acute and reversible TET deficiency and a sequencing method to map multiple DNA modifications and chromatin accessibility simultaneously on a single DNA molecule. Our research will provide crucial mechanistic insights into how TET and DNA modifications regulate gene expression and cell differentiation. Our findings will potentially identify genes involved in immunodeficiency and cancer, providing an opportunity to develop targeted treatments.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT Protein phosphorylation plays a key role in numerous cellular processes through a dynamic balance between protein kinases and phosphatases. In many disease conditions, this balance is inclined toward kinase hyperactivation and/or phosphatase inactivation, which are highly related to changes of their posttranslational modifications (PTMs). Being able to introduce multiple PTMs on such specific signaling enzymes in a chemically well-defined manner is both impactful and transformative to understand cell signaling pathways and the function of PTMs. In previous study, by employing protein semisynthesis methods to generate homogenous protein kinase Akt forms as substrates or stoichiometrically phosphorylated at Ser473 as calibration standards, we provided a detailed portrait of how mTOR Complex 2 (mTORC2) but not mTORC1 can selectively recognize and phosphorylate Akt Ser473 to activate this key signaling kinase. In this proposal, we continue to develop and utilize protein semisynthesis methods to elucidate the regulation of two key signaling enzymes including protein kinase S6K1 and the heterotrimeric phosphatase PP2A by PTMs and other allosteric mechanisms. S6K1 is a crucial downstream effector of mTORC1, and is critically regulated by the phosphorylation of a cluster of Ser/Thr residues (Ser411, Ser418, Thr421 and Ser424) in the C-terminal autoinhibitory domain (CTD). Yet how the CTD phosphorylation modulates S6K1 structure and function has been poorly defined. The PP2A phosphatase heterotrimer is responsible to the vast majority Ser/Thr phosphatase activity in eukaryotic cells, and its assembly has been linked to changes in the C-terminal PTMs of the C subunit including Thr304 and Tyr307 phosphorylations and Leu309 methylation. However, the function of these PTMs has yet to be fully characterized, and remain a great of interest in the field. It is also very little known how PP2A recruits its substrates, limiting our understanding of PP2A-regulated signaling. We will produce these two enzymes containing site-specific PTMs and their non-hydrolyzable analogs, and will integrate kinetic assays, structural analysis, binding measurements, and cell-based studies to clarify the structural and catalytic features. Moreover, we have developed a novel proximity crosslinking method using nanobodies as proximity-directing agents for analyzing PP2A interactome in different cellular conditions in response to various stimuli. Successful completion of this project will not only provide a detailed molecular understanding of how these two signaling enzymes regulated by specific PTMs, but inspire novel therapeutic strategies combat the diseases associated with their dysregulation.

NIH Research Projects · FY 2025 · 2023-09

Project Summary Lens shape is a primary determinant of its optical power and accommodative ability. Growth-induced change in lens shape directly causes presbyopia and is a known risk factor for age-related cataract. However, the biomechanical factors which interact to govern age-related changes in lens shape remain unknown. In the proposed study, we will systematically characterize the biomechanical properties of the aging human lens and use that information to understand how the lens shape evolves with age. A novel theory of constrained soft tissue growth will be employed to account for the presence of the lens capsule and its influence on the volumetric expansion of the lens fiber cell compartment. The biomechanical interaction between the fiber cell compartment and the capsule ultimately dictates the shape of the lens. These interactions behave like a water balloon filled with gelatin: as the gelatin sets, the properties of the gelatin contribute more and more to the shape of the balloon.